rabbit monoclonal anti naa10  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit monoclonal anti naa10
    ( A ) <t>Naa10</t> +/Y , Naa10 -/Y , and Naa10 tm1a/Y embryos at E10.5. Growth retardation (5/33, more than five somites lower or undersized compared to littermate controls), kinky trunk, and developmental arrest are shown in Naa10 -/Y (4/33) and Naa10 tm1a/Y (1/5). Scale bars: 500 μm. ( B ) The percentage lethality in newborns, comparing Naa10 wildtype (WT) ( Naa10 +/Y and Naa10 +/+ ), Naa10 -/+ and Naa10 -/Y pups until P3, derived from matings between heterozygous females and WT males. Approximately 11.6% (10/86) of WT, 24% (13/54) of Naa10 +/- , and 76.3% (29/38) of Naa10 -/Y mice were found dead before P3. ( C ) Representative images of Naa10 -/Y pups during early postnatal days compared with Naa10 +/Y . Severe developmental defects such as malformations of head and lower body (one leg; black arrowheads), whole-body edema, and anophthalmia (black arrows) are shown (N = 1 each). ( D ) Hematoxylin and eosin (H&E)-stained heart transverse section at E14.5 and vertical section at E18.5, comparing Naa10 +/Y and Naa10 -/Y embryos. Naa10 -/Y embryo shows a ventricular septal defect (VSD) at E14.5 and E18.5. Also, at E18.5, Naa10 -/Y embryo shows atrial septal defect (ASD). Arrow indicates VSD, ASD, and double outlet right ventricle (DORV). Scale bars: 20 μm.
    Rabbit Monoclonal Anti Naa10, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit monoclonal anti naa10/product/Cell Signaling Technology Inc
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit monoclonal anti naa10 - by Bioz Stars, 2023-02
    93/100 stars

    Images

    1) Product Images from "Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway"

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    Journal: eLife

    doi: 10.7554/eLife.65952

    ( A ) Naa10 +/Y , Naa10 -/Y , and Naa10 tm1a/Y embryos at E10.5. Growth retardation (5/33, more than five somites lower or undersized compared to littermate controls), kinky trunk, and developmental arrest are shown in Naa10 -/Y (4/33) and Naa10 tm1a/Y (1/5). Scale bars: 500 μm. ( B ) The percentage lethality in newborns, comparing Naa10 wildtype (WT) ( Naa10 +/Y and Naa10 +/+ ), Naa10 -/+ and Naa10 -/Y pups until P3, derived from matings between heterozygous females and WT males. Approximately 11.6% (10/86) of WT, 24% (13/54) of Naa10 +/- , and 76.3% (29/38) of Naa10 -/Y mice were found dead before P3. ( C ) Representative images of Naa10 -/Y pups during early postnatal days compared with Naa10 +/Y . Severe developmental defects such as malformations of head and lower body (one leg; black arrowheads), whole-body edema, and anophthalmia (black arrows) are shown (N = 1 each). ( D ) Hematoxylin and eosin (H&E)-stained heart transverse section at E14.5 and vertical section at E18.5, comparing Naa10 +/Y and Naa10 -/Y embryos. Naa10 -/Y embryo shows a ventricular septal defect (VSD) at E14.5 and E18.5. Also, at E18.5, Naa10 -/Y embryo shows atrial septal defect (ASD). Arrow indicates VSD, ASD, and double outlet right ventricle (DORV). Scale bars: 20 μm.
    Figure Legend Snippet: ( A ) Naa10 +/Y , Naa10 -/Y , and Naa10 tm1a/Y embryos at E10.5. Growth retardation (5/33, more than five somites lower or undersized compared to littermate controls), kinky trunk, and developmental arrest are shown in Naa10 -/Y (4/33) and Naa10 tm1a/Y (1/5). Scale bars: 500 μm. ( B ) The percentage lethality in newborns, comparing Naa10 wildtype (WT) ( Naa10 +/Y and Naa10 +/+ ), Naa10 -/+ and Naa10 -/Y pups until P3, derived from matings between heterozygous females and WT males. Approximately 11.6% (10/86) of WT, 24% (13/54) of Naa10 +/- , and 76.3% (29/38) of Naa10 -/Y mice were found dead before P3. ( C ) Representative images of Naa10 -/Y pups during early postnatal days compared with Naa10 +/Y . Severe developmental defects such as malformations of head and lower body (one leg; black arrowheads), whole-body edema, and anophthalmia (black arrows) are shown (N = 1 each). ( D ) Hematoxylin and eosin (H&E)-stained heart transverse section at E14.5 and vertical section at E18.5, comparing Naa10 +/Y and Naa10 -/Y embryos. Naa10 -/Y embryo shows a ventricular septal defect (VSD) at E14.5 and E18.5. Also, at E18.5, Naa10 -/Y embryo shows atrial septal defect (ASD). Arrow indicates VSD, ASD, and double outlet right ventricle (DORV). Scale bars: 20 μm.

    Techniques Used: Derivative Assay, Staining

    ( A ) Wildtype male heart outflow tract region indicating separate aorta and pulmonary trunks nestled between left and right atria. ( B ) Naa10 -/- female heart from dying P0 pup only has a single outflow tract emerging from the right ventricle, resulting in persistent truncus arteriosus. ( C ) Naa10 -/y male heart from dying P0 pup has separate outflow tracts but both emerge from the right ventricle, resulting in double outlet right ventricle. ( D ) Normal histology from wildtype male heart of pulmonary artery exiting the right ventricle. ( E ) Histology from Naa10 -/- heart of single outflow tract exiting the right ventricle with tricuspid valve leaflets. ( F ) Histology from Naa10 -/y heart of both pulmonary and aortic arteries emerging from right ventricle within the same plane. ( G ) Naa10 -/- histology showing membranous ventricular septal defect (VSD). ( H ) Naa10 -/y histology showing muscular VSD. ( I ) Normal heart revealing closed ductus arterious. ( J ) Naa10 -/- histology showing open ductus arterious leading to pulmonary overload and likely lethality.
    Figure Legend Snippet: ( A ) Wildtype male heart outflow tract region indicating separate aorta and pulmonary trunks nestled between left and right atria. ( B ) Naa10 -/- female heart from dying P0 pup only has a single outflow tract emerging from the right ventricle, resulting in persistent truncus arteriosus. ( C ) Naa10 -/y male heart from dying P0 pup has separate outflow tracts but both emerge from the right ventricle, resulting in double outlet right ventricle. ( D ) Normal histology from wildtype male heart of pulmonary artery exiting the right ventricle. ( E ) Histology from Naa10 -/- heart of single outflow tract exiting the right ventricle with tricuspid valve leaflets. ( F ) Histology from Naa10 -/y heart of both pulmonary and aortic arteries emerging from right ventricle within the same plane. ( G ) Naa10 -/- histology showing membranous ventricular septal defect (VSD). ( H ) Naa10 -/y histology showing muscular VSD. ( I ) Normal heart revealing closed ductus arterious. ( J ) Naa10 -/- histology showing open ductus arterious leading to pulmonary overload and likely lethality.

    Techniques Used:

    ( A–C ) Representative images of abnormalities in Naa10 -/Y compared with Naa10 +/Y . ( A ) Body weight of male (left) and female (right) versus ages was monitored from 2 weeks. The weight of Naa10 -/Y and Naa10 -/- mice is markedly reduced compared with that of the wildtype (WT) mice. Asterisks indicate a statistical difference calculated by Student’s t -test: *p<0.05. ( B ) Representative images of completely penetrant phenotypes. Hypopigmentation ( Naa10 +/Y , n = 243; Naa10 -/Y , n = 121; Naa10 tm1a/Y , n = 17) and supernumerary ribs ( Naa10 +/Y , n = 3; Naa10 -/Y , n = 6; Naa10 tm1a/Y , n = 2; E18.5) were found 100% in Naa10 -deficient mice. ( C ) Naa10 -/Y is smaller in size and has round-shaped head ( Naa10 +/Y 0/59, Naa10 -/Y 7/33). Over time, hydrocephaly became apparent (N = 14/29 [~48%] for >P7 male Naa10 -Y ; N = 7/19 [~36%] for >P7 female Naa10 -/- ). Hydronephrosis (red arrow, Naa10 +/Y 0/23, Naa10 -/Y 14/29, Naa10 +/+ 0/5, Naa10 -/- 7/19) and abnormal genitalia (black arrow) of male (middle, Naa10 +/Y 0/23, Naa10 -/Y 16/29) and female (bottom, hydrometrocolpos, Naa10 +/+ 0/5, Naa10 -/- 7/19) are shown.
    Figure Legend Snippet: ( A–C ) Representative images of abnormalities in Naa10 -/Y compared with Naa10 +/Y . ( A ) Body weight of male (left) and female (right) versus ages was monitored from 2 weeks. The weight of Naa10 -/Y and Naa10 -/- mice is markedly reduced compared with that of the wildtype (WT) mice. Asterisks indicate a statistical difference calculated by Student’s t -test: *p<0.05. ( B ) Representative images of completely penetrant phenotypes. Hypopigmentation ( Naa10 +/Y , n = 243; Naa10 -/Y , n = 121; Naa10 tm1a/Y , n = 17) and supernumerary ribs ( Naa10 +/Y , n = 3; Naa10 -/Y , n = 6; Naa10 tm1a/Y , n = 2; E18.5) were found 100% in Naa10 -deficient mice. ( C ) Naa10 -/Y is smaller in size and has round-shaped head ( Naa10 +/Y 0/59, Naa10 -/Y 7/33). Over time, hydrocephaly became apparent (N = 14/29 [~48%] for >P7 male Naa10 -Y ; N = 7/19 [~36%] for >P7 female Naa10 -/- ). Hydronephrosis (red arrow, Naa10 +/Y 0/23, Naa10 -/Y 14/29, Naa10 +/+ 0/5, Naa10 -/- 7/19) and abnormal genitalia (black arrow) of male (middle, Naa10 +/Y 0/23, Naa10 -/Y 16/29) and female (bottom, hydrometrocolpos, Naa10 +/+ 0/5, Naa10 -/- 7/19) are shown.

    Techniques Used:

    Skeletal analyses for ribs, sternebrae, and vertebrae.
    Figure Legend Snippet: Skeletal analyses for ribs, sternebrae, and vertebrae.

    Techniques Used:

    ( A ) In wildtype (WT) mice, 13 thoracic vertebrae and ribs are numbered, whereas 14 thoracic vertebrae and ribs are counted in mutants ( Naa10 -/Y ) (WT on the left, mutant on the right). n = 11 CT scans for Naa10 -/Y compared to n = 18 Naa10 +/Y . ( B–D ) Different number of ribs are linking the sternum between in Naa10 -/Y , Naa10 -/- , and WT. ( B ) Seven ribs linking the sternum in WT. ( C ) Eight ribs linking the sternum (the white arrow shows the eighth rib) in Naa10 -/Y . ( D ) Seven on one side plus seven and one almost linking on the other side. In two mice, an asymmetrical link was observed. White arrow shows the eighth asymmetrical rib. ( E, F ) Abnormalities in the cervical phenotype. ( E ) Cervical WT/morphology. ( F ) Partial fusion of C1 and C2 dorsal arch in one mutant mouse.
    Figure Legend Snippet: ( A ) In wildtype (WT) mice, 13 thoracic vertebrae and ribs are numbered, whereas 14 thoracic vertebrae and ribs are counted in mutants ( Naa10 -/Y ) (WT on the left, mutant on the right). n = 11 CT scans for Naa10 -/Y compared to n = 18 Naa10 +/Y . ( B–D ) Different number of ribs are linking the sternum between in Naa10 -/Y , Naa10 -/- , and WT. ( B ) Seven ribs linking the sternum in WT. ( C ) Eight ribs linking the sternum (the white arrow shows the eighth rib) in Naa10 -/Y . ( D ) Seven on one side plus seven and one almost linking on the other side. In two mice, an asymmetrical link was observed. White arrow shows the eighth asymmetrical rib. ( E, F ) Abnormalities in the cervical phenotype. ( E ) Cervical WT/morphology. ( F ) Partial fusion of C1 and C2 dorsal arch in one mutant mouse.

    Techniques Used: Mutagenesis

    ( A ) Representative images and histology of renal defects at E18.5 (n = 6 out of 39 examined) and P3 (n = 4 out of 11 examined). ( B ) Hydrocephaly (n = 3 CT scans for Naa10 -/Y mice with hydrocephaly compared to n = 3 Naa10 -/Y mice without hydrocephaly). ( C ) Kaplan–Meier survival curve of the Naa10 +/Y and Naa10 -/Y male mice starting at 4 days of life, thus not including any mice that died in the first three days of life.
    Figure Legend Snippet: ( A ) Representative images and histology of renal defects at E18.5 (n = 6 out of 39 examined) and P3 (n = 4 out of 11 examined). ( B ) Hydrocephaly (n = 3 CT scans for Naa10 -/Y mice with hydrocephaly compared to n = 3 Naa10 -/Y mice without hydrocephaly). ( C ) Kaplan–Meier survival curve of the Naa10 +/Y and Naa10 -/Y male mice starting at 4 days of life, thus not including any mice that died in the first three days of life.

    Techniques Used:

    ( A ) Correlation of Naa10 alteration state on amino-terminal acetylation in mouse embryonic fibroblasts (MEFs). Each dot (n = 533) represents the average amino-terminal acetylation percentage of five replicates of Naa10 +/Y and Naa10 -/Y , respectively. Dashed lines are the borders of ±10% difference. Except for the 10 dots, 522 of the 533 dots are within the borders. The marginal histograms show the distribution of amino-terminal acetylation data points. ( B ) Immunoprecipitation of Naa15. Liver tissue from WT and Naa10 KO mouse was lysed and incubated with anti-Naa15 antibody to retrieve NatA complexes. Proteins were separated by SDS-PAGE and immunoblots probed with anti-Naa15 antibody and anti-NAA10 antibody. ( C ) Catalytic activity of immunoprecipitated NatA. The catalytic activity of NatA precipitated from WT and Naa10 KO mouse liver tissue by anti-Naa15 was measured towards the NatA substrate peptide SESS 24 in an in vitro [ 14 C]-Ac-CoA–based acetylation assay. Control reactions were performed with no enzyme or no peptide to account for background signal. The immunoprecipitation (IP) and activity measurements were performed in three independent setups, each with three technical replicates per assay. One representative setup is shown. Figure 3—source data 1. Identification of a potential Naa10 homolog.
    Figure Legend Snippet: ( A ) Correlation of Naa10 alteration state on amino-terminal acetylation in mouse embryonic fibroblasts (MEFs). Each dot (n = 533) represents the average amino-terminal acetylation percentage of five replicates of Naa10 +/Y and Naa10 -/Y , respectively. Dashed lines are the borders of ±10% difference. Except for the 10 dots, 522 of the 533 dots are within the borders. The marginal histograms show the distribution of amino-terminal acetylation data points. ( B ) Immunoprecipitation of Naa15. Liver tissue from WT and Naa10 KO mouse was lysed and incubated with anti-Naa15 antibody to retrieve NatA complexes. Proteins were separated by SDS-PAGE and immunoblots probed with anti-Naa15 antibody and anti-NAA10 antibody. ( C ) Catalytic activity of immunoprecipitated NatA. The catalytic activity of NatA precipitated from WT and Naa10 KO mouse liver tissue by anti-Naa15 was measured towards the NatA substrate peptide SESS 24 in an in vitro [ 14 C]-Ac-CoA–based acetylation assay. Control reactions were performed with no enzyme or no peptide to account for background signal. The immunoprecipitation (IP) and activity measurements were performed in three independent setups, each with three technical replicates per assay. One representative setup is shown. Figure 3—source data 1. Identification of a potential Naa10 homolog.

    Techniques Used: Immunoprecipitation, Incubation, SDS Page, Western Blot, Activity Assay, In Vitro, Acetylation Assay

    ( A ) Expression of Naa10 and Naa12 in wildtype (WT), Naa10 KO and Naa12 KO tissues, adult mice 13 weeks of age, (brain, heart, kidney, and testis) by RT-PCR. Expression of GAPDH was analyzed as an endogenous control. ( B ) Phenotypes in Naa12 KO mice. Lack of hypopigmentation (upper; N = 29) and lack of supernumerary ribs (middle and bottom; E18.5; N = 7) in Naa12 KO mouse.
    Figure Legend Snippet: ( A ) Expression of Naa10 and Naa12 in wildtype (WT), Naa10 KO and Naa12 KO tissues, adult mice 13 weeks of age, (brain, heart, kidney, and testis) by RT-PCR. Expression of GAPDH was analyzed as an endogenous control. ( B ) Phenotypes in Naa12 KO mice. Lack of hypopigmentation (upper; N = 29) and lack of supernumerary ribs (middle and bottom; E18.5; N = 7) in Naa12 KO mouse.

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction

    ( A ) In vitro N-terminal acetyltransferase radioactive-based assay. Comparison of mouse Naa10 and Naa12 towards Naa10 peptide substrates, beta-actin (DDDIA-) and gamma-actin (EEEIA-), and the optimal NatA complex peptide substrate, SESSS-. Background control reactions were performed in the absence of either peptide or enzyme. Assays were performed in triplicate; error bars represent SEM. ( B ) Co-immunoprecipitation assay. HEK293 cells were transfected as indicated and lysed after 48 hr. Cell lysates were incubated with 1 μg anti-V5 antibody to precipitate V5-tagged Naa15. The isolated complexes were separated on SDS-PAGE and probed with the indicated antibodies. ( C ) Recombinant mouse Naa12/human Naa15 chimera complex activity. Radioactive acetyltransferase activity assay evaluating the activity of mNaa12-hNaa15 towards peptide (closed circles, ‘mNaa12-hNaa15’) and peptide chemical acetylation in the absence of enzyme (closed circles, ‘Buffer’) as well as chemical acetylation of the enzyme in the absence of peptide (open circles) assay and background (open circles). Error bars represent SD of two technical replicates. These are the same results from fraction #14 (both SESSS- and No Peptide) and both Buffer and Background used to illustrate the size-exclusion-purified mNaa12-hNaa15 complex activity in . Figure 4—source data 1. Characterization of a potential Naa10 homolog.
    Figure Legend Snippet: ( A ) In vitro N-terminal acetyltransferase radioactive-based assay. Comparison of mouse Naa10 and Naa12 towards Naa10 peptide substrates, beta-actin (DDDIA-) and gamma-actin (EEEIA-), and the optimal NatA complex peptide substrate, SESSS-. Background control reactions were performed in the absence of either peptide or enzyme. Assays were performed in triplicate; error bars represent SEM. ( B ) Co-immunoprecipitation assay. HEK293 cells were transfected as indicated and lysed after 48 hr. Cell lysates were incubated with 1 μg anti-V5 antibody to precipitate V5-tagged Naa15. The isolated complexes were separated on SDS-PAGE and probed with the indicated antibodies. ( C ) Recombinant mouse Naa12/human Naa15 chimera complex activity. Radioactive acetyltransferase activity assay evaluating the activity of mNaa12-hNaa15 towards peptide (closed circles, ‘mNaa12-hNaa15’) and peptide chemical acetylation in the absence of enzyme (closed circles, ‘Buffer’) as well as chemical acetylation of the enzyme in the absence of peptide (open circles) assay and background (open circles). Error bars represent SD of two technical replicates. These are the same results from fraction #14 (both SESSS- and No Peptide) and both Buffer and Background used to illustrate the size-exclusion-purified mNaa12-hNaa15 complex activity in . Figure 4—source data 1. Characterization of a potential Naa10 homolog.

    Techniques Used: In Vitro, Radioactivity, Co-Immunoprecipitation Assay, Transfection, Incubation, Isolation, SDS Page, Recombinant, Activity Assay, Purification

     Naa10,  Naa11, and Naa12 peptides identified by LC-MS/MS analysis in Naa15 IP samples from WT and Naa10 -KO mouse.
    Figure Legend Snippet: Naa10, Naa11, and Naa12 peptides identified by LC-MS/MS analysis in Naa15 IP samples from WT and Naa10 -KO mouse.

    Techniques Used: Sequencing

    Naa10 Naa12 DKO exhibit embryonic lethality. Pedigree and genotypes of pups and embryos at E10.5 and E18.5 from Naa10 +/- Naa12 +/- female mice crossed to the Naa10 +/Y Naa12 +/- male mice.
    Figure Legend Snippet: Naa10 Naa12 DKO exhibit embryonic lethality. Pedigree and genotypes of pups and embryos at E10.5 and E18.5 from Naa10 +/- Naa12 +/- female mice crossed to the Naa10 +/Y Naa12 +/- male mice.

    Techniques Used:

    Litter size of  Naa10  × Naa12 matings.
    Figure Legend Snippet: Litter size of Naa10 × Naa12 matings.

    Techniques Used:

    Naa10 Naa12 double-knockout (DKO) exhibit embryonic lethality. Pedigree of mating and genotypes of pups and embryos at E8.5, E10.5, E12.5, and E18.5.
    Figure Legend Snippet: Naa10 Naa12 double-knockout (DKO) exhibit embryonic lethality. Pedigree of mating and genotypes of pups and embryos at E8.5, E10.5, E12.5, and E18.5.

    Techniques Used: Double Knockout

    ( A ) Male body weight for the Naa10 mice on inbred genetic background (eight backcrosses to C57bl6/J). ( B ) Female body weight for the Naa10 mice on inbred genetic background (eight backcrosses to C57bl6/J). ( C ) Male body weight for the Naa10 and Naa12 mice on mixed genetic background. ( D ) Female body weight for the Naa10 and Naa12 mice on mixed genetic background.
    Figure Legend Snippet: ( A ) Male body weight for the Naa10 mice on inbred genetic background (eight backcrosses to C57bl6/J). ( B ) Female body weight for the Naa10 mice on inbred genetic background (eight backcrosses to C57bl6/J). ( C ) Male body weight for the Naa10 and Naa12 mice on mixed genetic background. ( D ) Female body weight for the Naa10 and Naa12 mice on mixed genetic background.

    Techniques Used:

    Model D 4 genotype survival by age.
    Figure Legend Snippet: Model D 4 genotype survival by age.

    Techniques Used:


    Figure Legend Snippet:

    Techniques Used: Isolation, Sequencing, Software

    rabbit monoclonal anti naa10  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
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  • 93

    Structured Review

    Cell Signaling Technology Inc rabbit monoclonal anti naa10
    ( A ) <t>Naa10</t> +/Y , Naa10 -/Y , and Naa10 tm1a/Y embryos at E10.5. Growth retardation (5/33, more than five somites lower or undersized compared to littermate controls), kinky trunk, and developmental arrest are shown in Naa10 -/Y (4/33) and Naa10 tm1a/Y (1/5). Scale bars: 500 μm. ( B ) The percentage lethality in newborns, comparing Naa10 wildtype (WT) ( Naa10 +/Y and Naa10 +/+ ), Naa10 -/+ and Naa10 -/Y pups until P3, derived from matings between heterozygous females and WT males. Approximately 11.6% (10/86) of WT, 24% (13/54) of Naa10 +/- , and 76.3% (29/38) of Naa10 -/Y mice were found dead before P3. ( C ) Representative images of Naa10 -/Y pups during early postnatal days compared with Naa10 +/Y . Severe developmental defects such as malformations of head and lower body (one leg; black arrowheads), whole-body edema, and anophthalmia (black arrows) are shown (N = 1 each). ( D ) Hematoxylin and eosin (H&E)-stained heart transverse section at E14.5 and vertical section at E18.5, comparing Naa10 +/Y and Naa10 -/Y embryos. Naa10 -/Y embryo shows a ventricular septal defect (VSD) at E14.5 and E18.5. Also, at E18.5, Naa10 -/Y embryo shows atrial septal defect (ASD). Arrow indicates VSD, ASD, and double outlet right ventricle (DORV). Scale bars: 20 μm.
    Rabbit Monoclonal Anti Naa10, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit monoclonal anti naa10/product/Cell Signaling Technology Inc
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit monoclonal anti naa10 - by Bioz Stars, 2023-02
    93/100 stars

    Images

    1) Product Images from "Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway"

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    Journal: eLife

    doi: 10.7554/eLife.65952

    ( A ) Naa10 +/Y , Naa10 -/Y , and Naa10 tm1a/Y embryos at E10.5. Growth retardation (5/33, more than five somites lower or undersized compared to littermate controls), kinky trunk, and developmental arrest are shown in Naa10 -/Y (4/33) and Naa10 tm1a/Y (1/5). Scale bars: 500 μm. ( B ) The percentage lethality in newborns, comparing Naa10 wildtype (WT) ( Naa10 +/Y and Naa10 +/+ ), Naa10 -/+ and Naa10 -/Y pups until P3, derived from matings between heterozygous females and WT males. Approximately 11.6% (10/86) of WT, 24% (13/54) of Naa10 +/- , and 76.3% (29/38) of Naa10 -/Y mice were found dead before P3. ( C ) Representative images of Naa10 -/Y pups during early postnatal days compared with Naa10 +/Y . Severe developmental defects such as malformations of head and lower body (one leg; black arrowheads), whole-body edema, and anophthalmia (black arrows) are shown (N = 1 each). ( D ) Hematoxylin and eosin (H&E)-stained heart transverse section at E14.5 and vertical section at E18.5, comparing Naa10 +/Y and Naa10 -/Y embryos. Naa10 -/Y embryo shows a ventricular septal defect (VSD) at E14.5 and E18.5. Also, at E18.5, Naa10 -/Y embryo shows atrial septal defect (ASD). Arrow indicates VSD, ASD, and double outlet right ventricle (DORV). Scale bars: 20 μm.
    Figure Legend Snippet: ( A ) Naa10 +/Y , Naa10 -/Y , and Naa10 tm1a/Y embryos at E10.5. Growth retardation (5/33, more than five somites lower or undersized compared to littermate controls), kinky trunk, and developmental arrest are shown in Naa10 -/Y (4/33) and Naa10 tm1a/Y (1/5). Scale bars: 500 μm. ( B ) The percentage lethality in newborns, comparing Naa10 wildtype (WT) ( Naa10 +/Y and Naa10 +/+ ), Naa10 -/+ and Naa10 -/Y pups until P3, derived from matings between heterozygous females and WT males. Approximately 11.6% (10/86) of WT, 24% (13/54) of Naa10 +/- , and 76.3% (29/38) of Naa10 -/Y mice were found dead before P3. ( C ) Representative images of Naa10 -/Y pups during early postnatal days compared with Naa10 +/Y . Severe developmental defects such as malformations of head and lower body (one leg; black arrowheads), whole-body edema, and anophthalmia (black arrows) are shown (N = 1 each). ( D ) Hematoxylin and eosin (H&E)-stained heart transverse section at E14.5 and vertical section at E18.5, comparing Naa10 +/Y and Naa10 -/Y embryos. Naa10 -/Y embryo shows a ventricular septal defect (VSD) at E14.5 and E18.5. Also, at E18.5, Naa10 -/Y embryo shows atrial septal defect (ASD). Arrow indicates VSD, ASD, and double outlet right ventricle (DORV). Scale bars: 20 μm.

    Techniques Used: Derivative Assay, Staining

    ( A ) Wildtype male heart outflow tract region indicating separate aorta and pulmonary trunks nestled between left and right atria. ( B ) Naa10 -/- female heart from dying P0 pup only has a single outflow tract emerging from the right ventricle, resulting in persistent truncus arteriosus. ( C ) Naa10 -/y male heart from dying P0 pup has separate outflow tracts but both emerge from the right ventricle, resulting in double outlet right ventricle. ( D ) Normal histology from wildtype male heart of pulmonary artery exiting the right ventricle. ( E ) Histology from Naa10 -/- heart of single outflow tract exiting the right ventricle with tricuspid valve leaflets. ( F ) Histology from Naa10 -/y heart of both pulmonary and aortic arteries emerging from right ventricle within the same plane. ( G ) Naa10 -/- histology showing membranous ventricular septal defect (VSD). ( H ) Naa10 -/y histology showing muscular VSD. ( I ) Normal heart revealing closed ductus arterious. ( J ) Naa10 -/- histology showing open ductus arterious leading to pulmonary overload and likely lethality.
    Figure Legend Snippet: ( A ) Wildtype male heart outflow tract region indicating separate aorta and pulmonary trunks nestled between left and right atria. ( B ) Naa10 -/- female heart from dying P0 pup only has a single outflow tract emerging from the right ventricle, resulting in persistent truncus arteriosus. ( C ) Naa10 -/y male heart from dying P0 pup has separate outflow tracts but both emerge from the right ventricle, resulting in double outlet right ventricle. ( D ) Normal histology from wildtype male heart of pulmonary artery exiting the right ventricle. ( E ) Histology from Naa10 -/- heart of single outflow tract exiting the right ventricle with tricuspid valve leaflets. ( F ) Histology from Naa10 -/y heart of both pulmonary and aortic arteries emerging from right ventricle within the same plane. ( G ) Naa10 -/- histology showing membranous ventricular septal defect (VSD). ( H ) Naa10 -/y histology showing muscular VSD. ( I ) Normal heart revealing closed ductus arterious. ( J ) Naa10 -/- histology showing open ductus arterious leading to pulmonary overload and likely lethality.

    Techniques Used:

    ( A–C ) Representative images of abnormalities in Naa10 -/Y compared with Naa10 +/Y . ( A ) Body weight of male (left) and female (right) versus ages was monitored from 2 weeks. The weight of Naa10 -/Y and Naa10 -/- mice is markedly reduced compared with that of the wildtype (WT) mice. Asterisks indicate a statistical difference calculated by Student’s t -test: *p<0.05. ( B ) Representative images of completely penetrant phenotypes. Hypopigmentation ( Naa10 +/Y , n = 243; Naa10 -/Y , n = 121; Naa10 tm1a/Y , n = 17) and supernumerary ribs ( Naa10 +/Y , n = 3; Naa10 -/Y , n = 6; Naa10 tm1a/Y , n = 2; E18.5) were found 100% in Naa10 -deficient mice. ( C ) Naa10 -/Y is smaller in size and has round-shaped head ( Naa10 +/Y 0/59, Naa10 -/Y 7/33). Over time, hydrocephaly became apparent (N = 14/29 [~48%] for >P7 male Naa10 -Y ; N = 7/19 [~36%] for >P7 female Naa10 -/- ). Hydronephrosis (red arrow, Naa10 +/Y 0/23, Naa10 -/Y 14/29, Naa10 +/+ 0/5, Naa10 -/- 7/19) and abnormal genitalia (black arrow) of male (middle, Naa10 +/Y 0/23, Naa10 -/Y 16/29) and female (bottom, hydrometrocolpos, Naa10 +/+ 0/5, Naa10 -/- 7/19) are shown.
    Figure Legend Snippet: ( A–C ) Representative images of abnormalities in Naa10 -/Y compared with Naa10 +/Y . ( A ) Body weight of male (left) and female (right) versus ages was monitored from 2 weeks. The weight of Naa10 -/Y and Naa10 -/- mice is markedly reduced compared with that of the wildtype (WT) mice. Asterisks indicate a statistical difference calculated by Student’s t -test: *p<0.05. ( B ) Representative images of completely penetrant phenotypes. Hypopigmentation ( Naa10 +/Y , n = 243; Naa10 -/Y , n = 121; Naa10 tm1a/Y , n = 17) and supernumerary ribs ( Naa10 +/Y , n = 3; Naa10 -/Y , n = 6; Naa10 tm1a/Y , n = 2; E18.5) were found 100% in Naa10 -deficient mice. ( C ) Naa10 -/Y is smaller in size and has round-shaped head ( Naa10 +/Y 0/59, Naa10 -/Y 7/33). Over time, hydrocephaly became apparent (N = 14/29 [~48%] for >P7 male Naa10 -Y ; N = 7/19 [~36%] for >P7 female Naa10 -/- ). Hydronephrosis (red arrow, Naa10 +/Y 0/23, Naa10 -/Y 14/29, Naa10 +/+ 0/5, Naa10 -/- 7/19) and abnormal genitalia (black arrow) of male (middle, Naa10 +/Y 0/23, Naa10 -/Y 16/29) and female (bottom, hydrometrocolpos, Naa10 +/+ 0/5, Naa10 -/- 7/19) are shown.

    Techniques Used:

    Skeletal analyses for ribs, sternebrae, and vertebrae.
    Figure Legend Snippet: Skeletal analyses for ribs, sternebrae, and vertebrae.

    Techniques Used:

    ( A ) In wildtype (WT) mice, 13 thoracic vertebrae and ribs are numbered, whereas 14 thoracic vertebrae and ribs are counted in mutants ( Naa10 -/Y ) (WT on the left, mutant on the right). n = 11 CT scans for Naa10 -/Y compared to n = 18 Naa10 +/Y . ( B–D ) Different number of ribs are linking the sternum between in Naa10 -/Y , Naa10 -/- , and WT. ( B ) Seven ribs linking the sternum in WT. ( C ) Eight ribs linking the sternum (the white arrow shows the eighth rib) in Naa10 -/Y . ( D ) Seven on one side plus seven and one almost linking on the other side. In two mice, an asymmetrical link was observed. White arrow shows the eighth asymmetrical rib. ( E, F ) Abnormalities in the cervical phenotype. ( E ) Cervical WT/morphology. ( F ) Partial fusion of C1 and C2 dorsal arch in one mutant mouse.
    Figure Legend Snippet: ( A ) In wildtype (WT) mice, 13 thoracic vertebrae and ribs are numbered, whereas 14 thoracic vertebrae and ribs are counted in mutants ( Naa10 -/Y ) (WT on the left, mutant on the right). n = 11 CT scans for Naa10 -/Y compared to n = 18 Naa10 +/Y . ( B–D ) Different number of ribs are linking the sternum between in Naa10 -/Y , Naa10 -/- , and WT. ( B ) Seven ribs linking the sternum in WT. ( C ) Eight ribs linking the sternum (the white arrow shows the eighth rib) in Naa10 -/Y . ( D ) Seven on one side plus seven and one almost linking on the other side. In two mice, an asymmetrical link was observed. White arrow shows the eighth asymmetrical rib. ( E, F ) Abnormalities in the cervical phenotype. ( E ) Cervical WT/morphology. ( F ) Partial fusion of C1 and C2 dorsal arch in one mutant mouse.

    Techniques Used: Mutagenesis

    ( A ) Representative images and histology of renal defects at E18.5 (n = 6 out of 39 examined) and P3 (n = 4 out of 11 examined). ( B ) Hydrocephaly (n = 3 CT scans for Naa10 -/Y mice with hydrocephaly compared to n = 3 Naa10 -/Y mice without hydrocephaly). ( C ) Kaplan–Meier survival curve of the Naa10 +/Y and Naa10 -/Y male mice starting at 4 days of life, thus not including any mice that died in the first three days of life.
    Figure Legend Snippet: ( A ) Representative images and histology of renal defects at E18.5 (n = 6 out of 39 examined) and P3 (n = 4 out of 11 examined). ( B ) Hydrocephaly (n = 3 CT scans for Naa10 -/Y mice with hydrocephaly compared to n = 3 Naa10 -/Y mice without hydrocephaly). ( C ) Kaplan–Meier survival curve of the Naa10 +/Y and Naa10 -/Y male mice starting at 4 days of life, thus not including any mice that died in the first three days of life.

    Techniques Used:

    ( A ) Correlation of Naa10 alteration state on amino-terminal acetylation in mouse embryonic fibroblasts (MEFs). Each dot (n = 533) represents the average amino-terminal acetylation percentage of five replicates of Naa10 +/Y and Naa10 -/Y , respectively. Dashed lines are the borders of ±10% difference. Except for the 10 dots, 522 of the 533 dots are within the borders. The marginal histograms show the distribution of amino-terminal acetylation data points. ( B ) Immunoprecipitation of Naa15. Liver tissue from WT and Naa10 KO mouse was lysed and incubated with anti-Naa15 antibody to retrieve NatA complexes. Proteins were separated by SDS-PAGE and immunoblots probed with anti-Naa15 antibody and anti-NAA10 antibody. ( C ) Catalytic activity of immunoprecipitated NatA. The catalytic activity of NatA precipitated from WT and Naa10 KO mouse liver tissue by anti-Naa15 was measured towards the NatA substrate peptide SESS 24 in an in vitro [ 14 C]-Ac-CoA–based acetylation assay. Control reactions were performed with no enzyme or no peptide to account for background signal. The immunoprecipitation (IP) and activity measurements were performed in three independent setups, each with three technical replicates per assay. One representative setup is shown. Figure 3—source data 1. Identification of a potential Naa10 homolog.
    Figure Legend Snippet: ( A ) Correlation of Naa10 alteration state on amino-terminal acetylation in mouse embryonic fibroblasts (MEFs). Each dot (n = 533) represents the average amino-terminal acetylation percentage of five replicates of Naa10 +/Y and Naa10 -/Y , respectively. Dashed lines are the borders of ±10% difference. Except for the 10 dots, 522 of the 533 dots are within the borders. The marginal histograms show the distribution of amino-terminal acetylation data points. ( B ) Immunoprecipitation of Naa15. Liver tissue from WT and Naa10 KO mouse was lysed and incubated with anti-Naa15 antibody to retrieve NatA complexes. Proteins were separated by SDS-PAGE and immunoblots probed with anti-Naa15 antibody and anti-NAA10 antibody. ( C ) Catalytic activity of immunoprecipitated NatA. The catalytic activity of NatA precipitated from WT and Naa10 KO mouse liver tissue by anti-Naa15 was measured towards the NatA substrate peptide SESS 24 in an in vitro [ 14 C]-Ac-CoA–based acetylation assay. Control reactions were performed with no enzyme or no peptide to account for background signal. The immunoprecipitation (IP) and activity measurements were performed in three independent setups, each with three technical replicates per assay. One representative setup is shown. Figure 3—source data 1. Identification of a potential Naa10 homolog.

    Techniques Used: Immunoprecipitation, Incubation, SDS Page, Western Blot, Activity Assay, In Vitro, Acetylation Assay

    ( A ) Expression of Naa10 and Naa12 in wildtype (WT), Naa10 KO and Naa12 KO tissues, adult mice 13 weeks of age, (brain, heart, kidney, and testis) by RT-PCR. Expression of GAPDH was analyzed as an endogenous control. ( B ) Phenotypes in Naa12 KO mice. Lack of hypopigmentation (upper; N = 29) and lack of supernumerary ribs (middle and bottom; E18.5; N = 7) in Naa12 KO mouse.
    Figure Legend Snippet: ( A ) Expression of Naa10 and Naa12 in wildtype (WT), Naa10 KO and Naa12 KO tissues, adult mice 13 weeks of age, (brain, heart, kidney, and testis) by RT-PCR. Expression of GAPDH was analyzed as an endogenous control. ( B ) Phenotypes in Naa12 KO mice. Lack of hypopigmentation (upper; N = 29) and lack of supernumerary ribs (middle and bottom; E18.5; N = 7) in Naa12 KO mouse.

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction

    ( A ) In vitro N-terminal acetyltransferase radioactive-based assay. Comparison of mouse Naa10 and Naa12 towards Naa10 peptide substrates, beta-actin (DDDIA-) and gamma-actin (EEEIA-), and the optimal NatA complex peptide substrate, SESSS-. Background control reactions were performed in the absence of either peptide or enzyme. Assays were performed in triplicate; error bars represent SEM. ( B ) Co-immunoprecipitation assay. HEK293 cells were transfected as indicated and lysed after 48 hr. Cell lysates were incubated with 1 μg anti-V5 antibody to precipitate V5-tagged Naa15. The isolated complexes were separated on SDS-PAGE and probed with the indicated antibodies. ( C ) Recombinant mouse Naa12/human Naa15 chimera complex activity. Radioactive acetyltransferase activity assay evaluating the activity of mNaa12-hNaa15 towards peptide (closed circles, ‘mNaa12-hNaa15’) and peptide chemical acetylation in the absence of enzyme (closed circles, ‘Buffer’) as well as chemical acetylation of the enzyme in the absence of peptide (open circles) assay and background (open circles). Error bars represent SD of two technical replicates. These are the same results from fraction #14 (both SESSS- and No Peptide) and both Buffer and Background used to illustrate the size-exclusion-purified mNaa12-hNaa15 complex activity in . Figure 4—source data 1. Characterization of a potential Naa10 homolog.
    Figure Legend Snippet: ( A ) In vitro N-terminal acetyltransferase radioactive-based assay. Comparison of mouse Naa10 and Naa12 towards Naa10 peptide substrates, beta-actin (DDDIA-) and gamma-actin (EEEIA-), and the optimal NatA complex peptide substrate, SESSS-. Background control reactions were performed in the absence of either peptide or enzyme. Assays were performed in triplicate; error bars represent SEM. ( B ) Co-immunoprecipitation assay. HEK293 cells were transfected as indicated and lysed after 48 hr. Cell lysates were incubated with 1 μg anti-V5 antibody to precipitate V5-tagged Naa15. The isolated complexes were separated on SDS-PAGE and probed with the indicated antibodies. ( C ) Recombinant mouse Naa12/human Naa15 chimera complex activity. Radioactive acetyltransferase activity assay evaluating the activity of mNaa12-hNaa15 towards peptide (closed circles, ‘mNaa12-hNaa15’) and peptide chemical acetylation in the absence of enzyme (closed circles, ‘Buffer’) as well as chemical acetylation of the enzyme in the absence of peptide (open circles) assay and background (open circles). Error bars represent SD of two technical replicates. These are the same results from fraction #14 (both SESSS- and No Peptide) and both Buffer and Background used to illustrate the size-exclusion-purified mNaa12-hNaa15 complex activity in . Figure 4—source data 1. Characterization of a potential Naa10 homolog.

    Techniques Used: In Vitro, Radioactivity, Co-Immunoprecipitation Assay, Transfection, Incubation, Isolation, SDS Page, Recombinant, Activity Assay, Purification

     Naa10,  Naa11, and Naa12 peptides identified by LC-MS/MS analysis in Naa15 IP samples from WT and Naa10 -KO mouse.
    Figure Legend Snippet: Naa10, Naa11, and Naa12 peptides identified by LC-MS/MS analysis in Naa15 IP samples from WT and Naa10 -KO mouse.

    Techniques Used: Sequencing

    Naa10 Naa12 DKO exhibit embryonic lethality. Pedigree and genotypes of pups and embryos at E10.5 and E18.5 from Naa10 +/- Naa12 +/- female mice crossed to the Naa10 +/Y Naa12 +/- male mice.
    Figure Legend Snippet: Naa10 Naa12 DKO exhibit embryonic lethality. Pedigree and genotypes of pups and embryos at E10.5 and E18.5 from Naa10 +/- Naa12 +/- female mice crossed to the Naa10 +/Y Naa12 +/- male mice.

    Techniques Used:

    Litter size of  Naa10  × Naa12 matings.
    Figure Legend Snippet: Litter size of Naa10 × Naa12 matings.

    Techniques Used:

    Naa10 Naa12 double-knockout (DKO) exhibit embryonic lethality. Pedigree of mating and genotypes of pups and embryos at E8.5, E10.5, E12.5, and E18.5.
    Figure Legend Snippet: Naa10 Naa12 double-knockout (DKO) exhibit embryonic lethality. Pedigree of mating and genotypes of pups and embryos at E8.5, E10.5, E12.5, and E18.5.

    Techniques Used: Double Knockout

    ( A ) Male body weight for the Naa10 mice on inbred genetic background (eight backcrosses to C57bl6/J). ( B ) Female body weight for the Naa10 mice on inbred genetic background (eight backcrosses to C57bl6/J). ( C ) Male body weight for the Naa10 and Naa12 mice on mixed genetic background. ( D ) Female body weight for the Naa10 and Naa12 mice on mixed genetic background.
    Figure Legend Snippet: ( A ) Male body weight for the Naa10 mice on inbred genetic background (eight backcrosses to C57bl6/J). ( B ) Female body weight for the Naa10 mice on inbred genetic background (eight backcrosses to C57bl6/J). ( C ) Male body weight for the Naa10 and Naa12 mice on mixed genetic background. ( D ) Female body weight for the Naa10 and Naa12 mice on mixed genetic background.

    Techniques Used:

    Model D 4 genotype survival by age.
    Figure Legend Snippet: Model D 4 genotype survival by age.

    Techniques Used:


    Figure Legend Snippet:

    Techniques Used: Isolation, Sequencing, Software

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    Cell Signaling Technology Inc anti naa10
    ( A ) <t>Naa10</t> +/Y , Naa10 -/Y , and Naa10 tm1a/Y embryos at E10.5. Growth retardation (5/33, more than five somites lower or undersized compared to littermate controls), kinky trunk, and developmental arrest are shown in Naa10 -/Y (4/33) and Naa10 tm1a/Y (1/5). Scale bars: 500 μm. ( B ) The percentage lethality in newborns, comparing Naa10 wildtype (WT) ( Naa10 +/Y and Naa10 +/+ ), Naa10 -/+ and Naa10 -/Y pups until P3, derived from matings between heterozygous females and WT males. Approximately 11.6% (10/86) of WT, 24% (13/54) of Naa10 +/- , and 76.3% (29/38) of Naa10 -/Y mice were found dead before P3. ( C ) Representative images of Naa10 -/Y pups during early postnatal days compared with Naa10 +/Y . Severe developmental defects such as malformations of head and lower body (one leg; black arrowheads), whole-body edema, and anophthalmia (black arrows) are shown (N = 1 each). ( D ) Hematoxylin and eosin (H&E)-stained heart transverse section at E14.5 and vertical section at E18.5, comparing Naa10 +/Y and Naa10 -/Y embryos. Naa10 -/Y embryo shows a ventricular septal defect (VSD) at E14.5 and E18.5. Also, at E18.5, Naa10 -/Y embryo shows atrial septal defect (ASD). Arrow indicates VSD, ASD, and double outlet right ventricle (DORV). Scale bars: 20 μm.
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    Images

    1) Product Images from "Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway"

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    Journal: eLife

    doi: 10.7554/eLife.65952

    ( A ) Naa10 +/Y , Naa10 -/Y , and Naa10 tm1a/Y embryos at E10.5. Growth retardation (5/33, more than five somites lower or undersized compared to littermate controls), kinky trunk, and developmental arrest are shown in Naa10 -/Y (4/33) and Naa10 tm1a/Y (1/5). Scale bars: 500 μm. ( B ) The percentage lethality in newborns, comparing Naa10 wildtype (WT) ( Naa10 +/Y and Naa10 +/+ ), Naa10 -/+ and Naa10 -/Y pups until P3, derived from matings between heterozygous females and WT males. Approximately 11.6% (10/86) of WT, 24% (13/54) of Naa10 +/- , and 76.3% (29/38) of Naa10 -/Y mice were found dead before P3. ( C ) Representative images of Naa10 -/Y pups during early postnatal days compared with Naa10 +/Y . Severe developmental defects such as malformations of head and lower body (one leg; black arrowheads), whole-body edema, and anophthalmia (black arrows) are shown (N = 1 each). ( D ) Hematoxylin and eosin (H&E)-stained heart transverse section at E14.5 and vertical section at E18.5, comparing Naa10 +/Y and Naa10 -/Y embryos. Naa10 -/Y embryo shows a ventricular septal defect (VSD) at E14.5 and E18.5. Also, at E18.5, Naa10 -/Y embryo shows atrial septal defect (ASD). Arrow indicates VSD, ASD, and double outlet right ventricle (DORV). Scale bars: 20 μm.
    Figure Legend Snippet: ( A ) Naa10 +/Y , Naa10 -/Y , and Naa10 tm1a/Y embryos at E10.5. Growth retardation (5/33, more than five somites lower or undersized compared to littermate controls), kinky trunk, and developmental arrest are shown in Naa10 -/Y (4/33) and Naa10 tm1a/Y (1/5). Scale bars: 500 μm. ( B ) The percentage lethality in newborns, comparing Naa10 wildtype (WT) ( Naa10 +/Y and Naa10 +/+ ), Naa10 -/+ and Naa10 -/Y pups until P3, derived from matings between heterozygous females and WT males. Approximately 11.6% (10/86) of WT, 24% (13/54) of Naa10 +/- , and 76.3% (29/38) of Naa10 -/Y mice were found dead before P3. ( C ) Representative images of Naa10 -/Y pups during early postnatal days compared with Naa10 +/Y . Severe developmental defects such as malformations of head and lower body (one leg; black arrowheads), whole-body edema, and anophthalmia (black arrows) are shown (N = 1 each). ( D ) Hematoxylin and eosin (H&E)-stained heart transverse section at E14.5 and vertical section at E18.5, comparing Naa10 +/Y and Naa10 -/Y embryos. Naa10 -/Y embryo shows a ventricular septal defect (VSD) at E14.5 and E18.5. Also, at E18.5, Naa10 -/Y embryo shows atrial septal defect (ASD). Arrow indicates VSD, ASD, and double outlet right ventricle (DORV). Scale bars: 20 μm.

    Techniques Used: Derivative Assay, Staining

    ( A ) Wildtype male heart outflow tract region indicating separate aorta and pulmonary trunks nestled between left and right atria. ( B ) Naa10 -/- female heart from dying P0 pup only has a single outflow tract emerging from the right ventricle, resulting in persistent truncus arteriosus. ( C ) Naa10 -/y male heart from dying P0 pup has separate outflow tracts but both emerge from the right ventricle, resulting in double outlet right ventricle. ( D ) Normal histology from wildtype male heart of pulmonary artery exiting the right ventricle. ( E ) Histology from Naa10 -/- heart of single outflow tract exiting the right ventricle with tricuspid valve leaflets. ( F ) Histology from Naa10 -/y heart of both pulmonary and aortic arteries emerging from right ventricle within the same plane. ( G ) Naa10 -/- histology showing membranous ventricular septal defect (VSD). ( H ) Naa10 -/y histology showing muscular VSD. ( I ) Normal heart revealing closed ductus arterious. ( J ) Naa10 -/- histology showing open ductus arterious leading to pulmonary overload and likely lethality.
    Figure Legend Snippet: ( A ) Wildtype male heart outflow tract region indicating separate aorta and pulmonary trunks nestled between left and right atria. ( B ) Naa10 -/- female heart from dying P0 pup only has a single outflow tract emerging from the right ventricle, resulting in persistent truncus arteriosus. ( C ) Naa10 -/y male heart from dying P0 pup has separate outflow tracts but both emerge from the right ventricle, resulting in double outlet right ventricle. ( D ) Normal histology from wildtype male heart of pulmonary artery exiting the right ventricle. ( E ) Histology from Naa10 -/- heart of single outflow tract exiting the right ventricle with tricuspid valve leaflets. ( F ) Histology from Naa10 -/y heart of both pulmonary and aortic arteries emerging from right ventricle within the same plane. ( G ) Naa10 -/- histology showing membranous ventricular septal defect (VSD). ( H ) Naa10 -/y histology showing muscular VSD. ( I ) Normal heart revealing closed ductus arterious. ( J ) Naa10 -/- histology showing open ductus arterious leading to pulmonary overload and likely lethality.

    Techniques Used:

    ( A–C ) Representative images of abnormalities in Naa10 -/Y compared with Naa10 +/Y . ( A ) Body weight of male (left) and female (right) versus ages was monitored from 2 weeks. The weight of Naa10 -/Y and Naa10 -/- mice is markedly reduced compared with that of the wildtype (WT) mice. Asterisks indicate a statistical difference calculated by Student’s t -test: *p<0.05. ( B ) Representative images of completely penetrant phenotypes. Hypopigmentation ( Naa10 +/Y , n = 243; Naa10 -/Y , n = 121; Naa10 tm1a/Y , n = 17) and supernumerary ribs ( Naa10 +/Y , n = 3; Naa10 -/Y , n = 6; Naa10 tm1a/Y , n = 2; E18.5) were found 100% in Naa10 -deficient mice. ( C ) Naa10 -/Y is smaller in size and has round-shaped head ( Naa10 +/Y 0/59, Naa10 -/Y 7/33). Over time, hydrocephaly became apparent (N = 14/29 [~48%] for >P7 male Naa10 -Y ; N = 7/19 [~36%] for >P7 female Naa10 -/- ). Hydronephrosis (red arrow, Naa10 +/Y 0/23, Naa10 -/Y 14/29, Naa10 +/+ 0/5, Naa10 -/- 7/19) and abnormal genitalia (black arrow) of male (middle, Naa10 +/Y 0/23, Naa10 -/Y 16/29) and female (bottom, hydrometrocolpos, Naa10 +/+ 0/5, Naa10 -/- 7/19) are shown.
    Figure Legend Snippet: ( A–C ) Representative images of abnormalities in Naa10 -/Y compared with Naa10 +/Y . ( A ) Body weight of male (left) and female (right) versus ages was monitored from 2 weeks. The weight of Naa10 -/Y and Naa10 -/- mice is markedly reduced compared with that of the wildtype (WT) mice. Asterisks indicate a statistical difference calculated by Student’s t -test: *p<0.05. ( B ) Representative images of completely penetrant phenotypes. Hypopigmentation ( Naa10 +/Y , n = 243; Naa10 -/Y , n = 121; Naa10 tm1a/Y , n = 17) and supernumerary ribs ( Naa10 +/Y , n = 3; Naa10 -/Y , n = 6; Naa10 tm1a/Y , n = 2; E18.5) were found 100% in Naa10 -deficient mice. ( C ) Naa10 -/Y is smaller in size and has round-shaped head ( Naa10 +/Y 0/59, Naa10 -/Y 7/33). Over time, hydrocephaly became apparent (N = 14/29 [~48%] for >P7 male Naa10 -Y ; N = 7/19 [~36%] for >P7 female Naa10 -/- ). Hydronephrosis (red arrow, Naa10 +/Y 0/23, Naa10 -/Y 14/29, Naa10 +/+ 0/5, Naa10 -/- 7/19) and abnormal genitalia (black arrow) of male (middle, Naa10 +/Y 0/23, Naa10 -/Y 16/29) and female (bottom, hydrometrocolpos, Naa10 +/+ 0/5, Naa10 -/- 7/19) are shown.

    Techniques Used:

    Skeletal analyses for ribs, sternebrae, and vertebrae.
    Figure Legend Snippet: Skeletal analyses for ribs, sternebrae, and vertebrae.

    Techniques Used:

    ( A ) In wildtype (WT) mice, 13 thoracic vertebrae and ribs are numbered, whereas 14 thoracic vertebrae and ribs are counted in mutants ( Naa10 -/Y ) (WT on the left, mutant on the right). n = 11 CT scans for Naa10 -/Y compared to n = 18 Naa10 +/Y . ( B–D ) Different number of ribs are linking the sternum between in Naa10 -/Y , Naa10 -/- , and WT. ( B ) Seven ribs linking the sternum in WT. ( C ) Eight ribs linking the sternum (the white arrow shows the eighth rib) in Naa10 -/Y . ( D ) Seven on one side plus seven and one almost linking on the other side. In two mice, an asymmetrical link was observed. White arrow shows the eighth asymmetrical rib. ( E, F ) Abnormalities in the cervical phenotype. ( E ) Cervical WT/morphology. ( F ) Partial fusion of C1 and C2 dorsal arch in one mutant mouse.
    Figure Legend Snippet: ( A ) In wildtype (WT) mice, 13 thoracic vertebrae and ribs are numbered, whereas 14 thoracic vertebrae and ribs are counted in mutants ( Naa10 -/Y ) (WT on the left, mutant on the right). n = 11 CT scans for Naa10 -/Y compared to n = 18 Naa10 +/Y . ( B–D ) Different number of ribs are linking the sternum between in Naa10 -/Y , Naa10 -/- , and WT. ( B ) Seven ribs linking the sternum in WT. ( C ) Eight ribs linking the sternum (the white arrow shows the eighth rib) in Naa10 -/Y . ( D ) Seven on one side plus seven and one almost linking on the other side. In two mice, an asymmetrical link was observed. White arrow shows the eighth asymmetrical rib. ( E, F ) Abnormalities in the cervical phenotype. ( E ) Cervical WT/morphology. ( F ) Partial fusion of C1 and C2 dorsal arch in one mutant mouse.

    Techniques Used: Mutagenesis

    ( A ) Representative images and histology of renal defects at E18.5 (n = 6 out of 39 examined) and P3 (n = 4 out of 11 examined). ( B ) Hydrocephaly (n = 3 CT scans for Naa10 -/Y mice with hydrocephaly compared to n = 3 Naa10 -/Y mice without hydrocephaly). ( C ) Kaplan–Meier survival curve of the Naa10 +/Y and Naa10 -/Y male mice starting at 4 days of life, thus not including any mice that died in the first three days of life.
    Figure Legend Snippet: ( A ) Representative images and histology of renal defects at E18.5 (n = 6 out of 39 examined) and P3 (n = 4 out of 11 examined). ( B ) Hydrocephaly (n = 3 CT scans for Naa10 -/Y mice with hydrocephaly compared to n = 3 Naa10 -/Y mice without hydrocephaly). ( C ) Kaplan–Meier survival curve of the Naa10 +/Y and Naa10 -/Y male mice starting at 4 days of life, thus not including any mice that died in the first three days of life.

    Techniques Used:

    ( A ) Correlation of Naa10 alteration state on amino-terminal acetylation in mouse embryonic fibroblasts (MEFs). Each dot (n = 533) represents the average amino-terminal acetylation percentage of five replicates of Naa10 +/Y and Naa10 -/Y , respectively. Dashed lines are the borders of ±10% difference. Except for the 10 dots, 522 of the 533 dots are within the borders. The marginal histograms show the distribution of amino-terminal acetylation data points. ( B ) Immunoprecipitation of Naa15. Liver tissue from WT and Naa10 KO mouse was lysed and incubated with anti-Naa15 antibody to retrieve NatA complexes. Proteins were separated by SDS-PAGE and immunoblots probed with anti-Naa15 antibody and anti-NAA10 antibody. ( C ) Catalytic activity of immunoprecipitated NatA. The catalytic activity of NatA precipitated from WT and Naa10 KO mouse liver tissue by anti-Naa15 was measured towards the NatA substrate peptide SESS 24 in an in vitro [ 14 C]-Ac-CoA–based acetylation assay. Control reactions were performed with no enzyme or no peptide to account for background signal. The immunoprecipitation (IP) and activity measurements were performed in three independent setups, each with three technical replicates per assay. One representative setup is shown. Figure 3—source data 1. Identification of a potential Naa10 homolog.
    Figure Legend Snippet: ( A ) Correlation of Naa10 alteration state on amino-terminal acetylation in mouse embryonic fibroblasts (MEFs). Each dot (n = 533) represents the average amino-terminal acetylation percentage of five replicates of Naa10 +/Y and Naa10 -/Y , respectively. Dashed lines are the borders of ±10% difference. Except for the 10 dots, 522 of the 533 dots are within the borders. The marginal histograms show the distribution of amino-terminal acetylation data points. ( B ) Immunoprecipitation of Naa15. Liver tissue from WT and Naa10 KO mouse was lysed and incubated with anti-Naa15 antibody to retrieve NatA complexes. Proteins were separated by SDS-PAGE and immunoblots probed with anti-Naa15 antibody and anti-NAA10 antibody. ( C ) Catalytic activity of immunoprecipitated NatA. The catalytic activity of NatA precipitated from WT and Naa10 KO mouse liver tissue by anti-Naa15 was measured towards the NatA substrate peptide SESS 24 in an in vitro [ 14 C]-Ac-CoA–based acetylation assay. Control reactions were performed with no enzyme or no peptide to account for background signal. The immunoprecipitation (IP) and activity measurements were performed in three independent setups, each with three technical replicates per assay. One representative setup is shown. Figure 3—source data 1. Identification of a potential Naa10 homolog.

    Techniques Used: Immunoprecipitation, Incubation, SDS Page, Western Blot, Activity Assay, In Vitro, Acetylation Assay

    ( A ) Expression of Naa10 and Naa12 in wildtype (WT), Naa10 KO and Naa12 KO tissues, adult mice 13 weeks of age, (brain, heart, kidney, and testis) by RT-PCR. Expression of GAPDH was analyzed as an endogenous control. ( B ) Phenotypes in Naa12 KO mice. Lack of hypopigmentation (upper; N = 29) and lack of supernumerary ribs (middle and bottom; E18.5; N = 7) in Naa12 KO mouse.
    Figure Legend Snippet: ( A ) Expression of Naa10 and Naa12 in wildtype (WT), Naa10 KO and Naa12 KO tissues, adult mice 13 weeks of age, (brain, heart, kidney, and testis) by RT-PCR. Expression of GAPDH was analyzed as an endogenous control. ( B ) Phenotypes in Naa12 KO mice. Lack of hypopigmentation (upper; N = 29) and lack of supernumerary ribs (middle and bottom; E18.5; N = 7) in Naa12 KO mouse.

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction

    ( A ) In vitro N-terminal acetyltransferase radioactive-based assay. Comparison of mouse Naa10 and Naa12 towards Naa10 peptide substrates, beta-actin (DDDIA-) and gamma-actin (EEEIA-), and the optimal NatA complex peptide substrate, SESSS-. Background control reactions were performed in the absence of either peptide or enzyme. Assays were performed in triplicate; error bars represent SEM. ( B ) Co-immunoprecipitation assay. HEK293 cells were transfected as indicated and lysed after 48 hr. Cell lysates were incubated with 1 μg anti-V5 antibody to precipitate V5-tagged Naa15. The isolated complexes were separated on SDS-PAGE and probed with the indicated antibodies. ( C ) Recombinant mouse Naa12/human Naa15 chimera complex activity. Radioactive acetyltransferase activity assay evaluating the activity of mNaa12-hNaa15 towards peptide (closed circles, ‘mNaa12-hNaa15’) and peptide chemical acetylation in the absence of enzyme (closed circles, ‘Buffer’) as well as chemical acetylation of the enzyme in the absence of peptide (open circles) assay and background (open circles). Error bars represent SD of two technical replicates. These are the same results from fraction #14 (both SESSS- and No Peptide) and both Buffer and Background used to illustrate the size-exclusion-purified mNaa12-hNaa15 complex activity in . Figure 4—source data 1. Characterization of a potential Naa10 homolog.
    Figure Legend Snippet: ( A ) In vitro N-terminal acetyltransferase radioactive-based assay. Comparison of mouse Naa10 and Naa12 towards Naa10 peptide substrates, beta-actin (DDDIA-) and gamma-actin (EEEIA-), and the optimal NatA complex peptide substrate, SESSS-. Background control reactions were performed in the absence of either peptide or enzyme. Assays were performed in triplicate; error bars represent SEM. ( B ) Co-immunoprecipitation assay. HEK293 cells were transfected as indicated and lysed after 48 hr. Cell lysates were incubated with 1 μg anti-V5 antibody to precipitate V5-tagged Naa15. The isolated complexes were separated on SDS-PAGE and probed with the indicated antibodies. ( C ) Recombinant mouse Naa12/human Naa15 chimera complex activity. Radioactive acetyltransferase activity assay evaluating the activity of mNaa12-hNaa15 towards peptide (closed circles, ‘mNaa12-hNaa15’) and peptide chemical acetylation in the absence of enzyme (closed circles, ‘Buffer’) as well as chemical acetylation of the enzyme in the absence of peptide (open circles) assay and background (open circles). Error bars represent SD of two technical replicates. These are the same results from fraction #14 (both SESSS- and No Peptide) and both Buffer and Background used to illustrate the size-exclusion-purified mNaa12-hNaa15 complex activity in . Figure 4—source data 1. Characterization of a potential Naa10 homolog.

    Techniques Used: In Vitro, Radioactivity, Co-Immunoprecipitation Assay, Transfection, Incubation, Isolation, SDS Page, Recombinant, Activity Assay, Purification

     Naa10,  Naa11, and Naa12 peptides identified by LC-MS/MS analysis in Naa15 IP samples from WT and Naa10 -KO mouse.
    Figure Legend Snippet: Naa10, Naa11, and Naa12 peptides identified by LC-MS/MS analysis in Naa15 IP samples from WT and Naa10 -KO mouse.

    Techniques Used: Sequencing

    Naa10 Naa12 DKO exhibit embryonic lethality. Pedigree and genotypes of pups and embryos at E10.5 and E18.5 from Naa10 +/- Naa12 +/- female mice crossed to the Naa10 +/Y Naa12 +/- male mice.
    Figure Legend Snippet: Naa10 Naa12 DKO exhibit embryonic lethality. Pedigree and genotypes of pups and embryos at E10.5 and E18.5 from Naa10 +/- Naa12 +/- female mice crossed to the Naa10 +/Y Naa12 +/- male mice.

    Techniques Used:

    Litter size of  Naa10  × Naa12 matings.
    Figure Legend Snippet: Litter size of Naa10 × Naa12 matings.

    Techniques Used:

    Naa10 Naa12 double-knockout (DKO) exhibit embryonic lethality. Pedigree of mating and genotypes of pups and embryos at E8.5, E10.5, E12.5, and E18.5.
    Figure Legend Snippet: Naa10 Naa12 double-knockout (DKO) exhibit embryonic lethality. Pedigree of mating and genotypes of pups and embryos at E8.5, E10.5, E12.5, and E18.5.

    Techniques Used: Double Knockout

    ( A ) Male body weight for the Naa10 mice on inbred genetic background (eight backcrosses to C57bl6/J). ( B ) Female body weight for the Naa10 mice on inbred genetic background (eight backcrosses to C57bl6/J). ( C ) Male body weight for the Naa10 and Naa12 mice on mixed genetic background. ( D ) Female body weight for the Naa10 and Naa12 mice on mixed genetic background.
    Figure Legend Snippet: ( A ) Male body weight for the Naa10 mice on inbred genetic background (eight backcrosses to C57bl6/J). ( B ) Female body weight for the Naa10 mice on inbred genetic background (eight backcrosses to C57bl6/J). ( C ) Male body weight for the Naa10 and Naa12 mice on mixed genetic background. ( D ) Female body weight for the Naa10 and Naa12 mice on mixed genetic background.

    Techniques Used:

    Model D 4 genotype survival by age.
    Figure Legend Snippet: Model D 4 genotype survival by age.

    Techniques Used:


    Figure Legend Snippet:

    Techniques Used: Isolation, Sequencing, Software

    rabbit monoclonal anti naa10  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit monoclonal anti naa10
    ( A ) <t>Naa10</t> +/Y , Naa10 -/Y , and Naa10 tm1a/Y embryos at E10.5. Growth retardation (5/33, more than five somites lower or undersized compared to littermate controls), kinky trunk, and developmental arrest are shown in Naa10 -/Y (4/33) and Naa10 tm1a/Y (1/5). Scale bars: 500 μm. ( B ) The percentage lethality in newborns, comparing Naa10 wildtype (WT) ( Naa10 +/Y and Naa10 +/+ ), Naa10 -/+ and Naa10 -/Y pups until P3, derived from matings between heterozygous females and WT males. Approximately 11.6% (10/86) of WT, 24% (13/54) of Naa10 +/- , and 76.3% (29/38) of Naa10 -/Y mice were found dead before P3. ( C ) Representative images of Naa10 -/Y pups during early postnatal days compared with Naa10 +/Y . Severe developmental defects such as malformations of head and lower body (one leg; black arrowheads), whole-body edema, and anophthalmia (black arrows) are shown (N = 1 each). ( D ) Hematoxylin and eosin (H&E)-stained heart transverse section at E14.5 and vertical section at E18.5, comparing Naa10 +/Y and Naa10 -/Y embryos. Naa10 -/Y embryo shows a ventricular septal defect (VSD) at E14.5 and E18.5. Also, at E18.5, Naa10 -/Y embryo shows atrial septal defect (ASD). Arrow indicates VSD, ASD, and double outlet right ventricle (DORV). Scale bars: 20 μm.
    Rabbit Monoclonal Anti Naa10, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway"

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    Journal: eLife

    doi: 10.7554/eLife.65952

    ( A ) Naa10 +/Y , Naa10 -/Y , and Naa10 tm1a/Y embryos at E10.5. Growth retardation (5/33, more than five somites lower or undersized compared to littermate controls), kinky trunk, and developmental arrest are shown in Naa10 -/Y (4/33) and Naa10 tm1a/Y (1/5). Scale bars: 500 μm. ( B ) The percentage lethality in newborns, comparing Naa10 wildtype (WT) ( Naa10 +/Y and Naa10 +/+ ), Naa10 -/+ and Naa10 -/Y pups until P3, derived from matings between heterozygous females and WT males. Approximately 11.6% (10/86) of WT, 24% (13/54) of Naa10 +/- , and 76.3% (29/38) of Naa10 -/Y mice were found dead before P3. ( C ) Representative images of Naa10 -/Y pups during early postnatal days compared with Naa10 +/Y . Severe developmental defects such as malformations of head and lower body (one leg; black arrowheads), whole-body edema, and anophthalmia (black arrows) are shown (N = 1 each). ( D ) Hematoxylin and eosin (H&E)-stained heart transverse section at E14.5 and vertical section at E18.5, comparing Naa10 +/Y and Naa10 -/Y embryos. Naa10 -/Y embryo shows a ventricular septal defect (VSD) at E14.5 and E18.5. Also, at E18.5, Naa10 -/Y embryo shows atrial septal defect (ASD). Arrow indicates VSD, ASD, and double outlet right ventricle (DORV). Scale bars: 20 μm.
    Figure Legend Snippet: ( A ) Naa10 +/Y , Naa10 -/Y , and Naa10 tm1a/Y embryos at E10.5. Growth retardation (5/33, more than five somites lower or undersized compared to littermate controls), kinky trunk, and developmental arrest are shown in Naa10 -/Y (4/33) and Naa10 tm1a/Y (1/5). Scale bars: 500 μm. ( B ) The percentage lethality in newborns, comparing Naa10 wildtype (WT) ( Naa10 +/Y and Naa10 +/+ ), Naa10 -/+ and Naa10 -/Y pups until P3, derived from matings between heterozygous females and WT males. Approximately 11.6% (10/86) of WT, 24% (13/54) of Naa10 +/- , and 76.3% (29/38) of Naa10 -/Y mice were found dead before P3. ( C ) Representative images of Naa10 -/Y pups during early postnatal days compared with Naa10 +/Y . Severe developmental defects such as malformations of head and lower body (one leg; black arrowheads), whole-body edema, and anophthalmia (black arrows) are shown (N = 1 each). ( D ) Hematoxylin and eosin (H&E)-stained heart transverse section at E14.5 and vertical section at E18.5, comparing Naa10 +/Y and Naa10 -/Y embryos. Naa10 -/Y embryo shows a ventricular septal defect (VSD) at E14.5 and E18.5. Also, at E18.5, Naa10 -/Y embryo shows atrial septal defect (ASD). Arrow indicates VSD, ASD, and double outlet right ventricle (DORV). Scale bars: 20 μm.

    Techniques Used: Derivative Assay, Staining

    ( A ) Wildtype male heart outflow tract region indicating separate aorta and pulmonary trunks nestled between left and right atria. ( B ) Naa10 -/- female heart from dying P0 pup only has a single outflow tract emerging from the right ventricle, resulting in persistent truncus arteriosus. ( C ) Naa10 -/y male heart from dying P0 pup has separate outflow tracts but both emerge from the right ventricle, resulting in double outlet right ventricle. ( D ) Normal histology from wildtype male heart of pulmonary artery exiting the right ventricle. ( E ) Histology from Naa10 -/- heart of single outflow tract exiting the right ventricle with tricuspid valve leaflets. ( F ) Histology from Naa10 -/y heart of both pulmonary and aortic arteries emerging from right ventricle within the same plane. ( G ) Naa10 -/- histology showing membranous ventricular septal defect (VSD). ( H ) Naa10 -/y histology showing muscular VSD. ( I ) Normal heart revealing closed ductus arterious. ( J ) Naa10 -/- histology showing open ductus arterious leading to pulmonary overload and likely lethality.
    Figure Legend Snippet: ( A ) Wildtype male heart outflow tract region indicating separate aorta and pulmonary trunks nestled between left and right atria. ( B ) Naa10 -/- female heart from dying P0 pup only has a single outflow tract emerging from the right ventricle, resulting in persistent truncus arteriosus. ( C ) Naa10 -/y male heart from dying P0 pup has separate outflow tracts but both emerge from the right ventricle, resulting in double outlet right ventricle. ( D ) Normal histology from wildtype male heart of pulmonary artery exiting the right ventricle. ( E ) Histology from Naa10 -/- heart of single outflow tract exiting the right ventricle with tricuspid valve leaflets. ( F ) Histology from Naa10 -/y heart of both pulmonary and aortic arteries emerging from right ventricle within the same plane. ( G ) Naa10 -/- histology showing membranous ventricular septal defect (VSD). ( H ) Naa10 -/y histology showing muscular VSD. ( I ) Normal heart revealing closed ductus arterious. ( J ) Naa10 -/- histology showing open ductus arterious leading to pulmonary overload and likely lethality.

    Techniques Used:

    ( A–C ) Representative images of abnormalities in Naa10 -/Y compared with Naa10 +/Y . ( A ) Body weight of male (left) and female (right) versus ages was monitored from 2 weeks. The weight of Naa10 -/Y and Naa10 -/- mice is markedly reduced compared with that of the wildtype (WT) mice. Asterisks indicate a statistical difference calculated by Student’s t -test: *p<0.05. ( B ) Representative images of completely penetrant phenotypes. Hypopigmentation ( Naa10 +/Y , n = 243; Naa10 -/Y , n = 121; Naa10 tm1a/Y , n = 17) and supernumerary ribs ( Naa10 +/Y , n = 3; Naa10 -/Y , n = 6; Naa10 tm1a/Y , n = 2; E18.5) were found 100% in Naa10 -deficient mice. ( C ) Naa10 -/Y is smaller in size and has round-shaped head ( Naa10 +/Y 0/59, Naa10 -/Y 7/33). Over time, hydrocephaly became apparent (N = 14/29 [~48%] for >P7 male Naa10 -Y ; N = 7/19 [~36%] for >P7 female Naa10 -/- ). Hydronephrosis (red arrow, Naa10 +/Y 0/23, Naa10 -/Y 14/29, Naa10 +/+ 0/5, Naa10 -/- 7/19) and abnormal genitalia (black arrow) of male (middle, Naa10 +/Y 0/23, Naa10 -/Y 16/29) and female (bottom, hydrometrocolpos, Naa10 +/+ 0/5, Naa10 -/- 7/19) are shown.
    Figure Legend Snippet: ( A–C ) Representative images of abnormalities in Naa10 -/Y compared with Naa10 +/Y . ( A ) Body weight of male (left) and female (right) versus ages was monitored from 2 weeks. The weight of Naa10 -/Y and Naa10 -/- mice is markedly reduced compared with that of the wildtype (WT) mice. Asterisks indicate a statistical difference calculated by Student’s t -test: *p<0.05. ( B ) Representative images of completely penetrant phenotypes. Hypopigmentation ( Naa10 +/Y , n = 243; Naa10 -/Y , n = 121; Naa10 tm1a/Y , n = 17) and supernumerary ribs ( Naa10 +/Y , n = 3; Naa10 -/Y , n = 6; Naa10 tm1a/Y , n = 2; E18.5) were found 100% in Naa10 -deficient mice. ( C ) Naa10 -/Y is smaller in size and has round-shaped head ( Naa10 +/Y 0/59, Naa10 -/Y 7/33). Over time, hydrocephaly became apparent (N = 14/29 [~48%] for >P7 male Naa10 -Y ; N = 7/19 [~36%] for >P7 female Naa10 -/- ). Hydronephrosis (red arrow, Naa10 +/Y 0/23, Naa10 -/Y 14/29, Naa10 +/+ 0/5, Naa10 -/- 7/19) and abnormal genitalia (black arrow) of male (middle, Naa10 +/Y 0/23, Naa10 -/Y 16/29) and female (bottom, hydrometrocolpos, Naa10 +/+ 0/5, Naa10 -/- 7/19) are shown.

    Techniques Used:

    Skeletal analyses for ribs, sternebrae, and vertebrae.
    Figure Legend Snippet: Skeletal analyses for ribs, sternebrae, and vertebrae.

    Techniques Used:

    ( A ) In wildtype (WT) mice, 13 thoracic vertebrae and ribs are numbered, whereas 14 thoracic vertebrae and ribs are counted in mutants ( Naa10 -/Y ) (WT on the left, mutant on the right). n = 11 CT scans for Naa10 -/Y compared to n = 18 Naa10 +/Y . ( B–D ) Different number of ribs are linking the sternum between in Naa10 -/Y , Naa10 -/- , and WT. ( B ) Seven ribs linking the sternum in WT. ( C ) Eight ribs linking the sternum (the white arrow shows the eighth rib) in Naa10 -/Y . ( D ) Seven on one side plus seven and one almost linking on the other side. In two mice, an asymmetrical link was observed. White arrow shows the eighth asymmetrical rib. ( E, F ) Abnormalities in the cervical phenotype. ( E ) Cervical WT/morphology. ( F ) Partial fusion of C1 and C2 dorsal arch in one mutant mouse.
    Figure Legend Snippet: ( A ) In wildtype (WT) mice, 13 thoracic vertebrae and ribs are numbered, whereas 14 thoracic vertebrae and ribs are counted in mutants ( Naa10 -/Y ) (WT on the left, mutant on the right). n = 11 CT scans for Naa10 -/Y compared to n = 18 Naa10 +/Y . ( B–D ) Different number of ribs are linking the sternum between in Naa10 -/Y , Naa10 -/- , and WT. ( B ) Seven ribs linking the sternum in WT. ( C ) Eight ribs linking the sternum (the white arrow shows the eighth rib) in Naa10 -/Y . ( D ) Seven on one side plus seven and one almost linking on the other side. In two mice, an asymmetrical link was observed. White arrow shows the eighth asymmetrical rib. ( E, F ) Abnormalities in the cervical phenotype. ( E ) Cervical WT/morphology. ( F ) Partial fusion of C1 and C2 dorsal arch in one mutant mouse.

    Techniques Used: Mutagenesis

    ( A ) Representative images and histology of renal defects at E18.5 (n = 6 out of 39 examined) and P3 (n = 4 out of 11 examined). ( B ) Hydrocephaly (n = 3 CT scans for Naa10 -/Y mice with hydrocephaly compared to n = 3 Naa10 -/Y mice without hydrocephaly). ( C ) Kaplan–Meier survival curve of the Naa10 +/Y and Naa10 -/Y male mice starting at 4 days of life, thus not including any mice that died in the first three days of life.
    Figure Legend Snippet: ( A ) Representative images and histology of renal defects at E18.5 (n = 6 out of 39 examined) and P3 (n = 4 out of 11 examined). ( B ) Hydrocephaly (n = 3 CT scans for Naa10 -/Y mice with hydrocephaly compared to n = 3 Naa10 -/Y mice without hydrocephaly). ( C ) Kaplan–Meier survival curve of the Naa10 +/Y and Naa10 -/Y male mice starting at 4 days of life, thus not including any mice that died in the first three days of life.

    Techniques Used:

    ( A ) Correlation of Naa10 alteration state on amino-terminal acetylation in mouse embryonic fibroblasts (MEFs). Each dot (n = 533) represents the average amino-terminal acetylation percentage of five replicates of Naa10 +/Y and Naa10 -/Y , respectively. Dashed lines are the borders of ±10% difference. Except for the 10 dots, 522 of the 533 dots are within the borders. The marginal histograms show the distribution of amino-terminal acetylation data points. ( B ) Immunoprecipitation of Naa15. Liver tissue from WT and Naa10 KO mouse was lysed and incubated with anti-Naa15 antibody to retrieve NatA complexes. Proteins were separated by SDS-PAGE and immunoblots probed with anti-Naa15 antibody and anti-NAA10 antibody. ( C ) Catalytic activity of immunoprecipitated NatA. The catalytic activity of NatA precipitated from WT and Naa10 KO mouse liver tissue by anti-Naa15 was measured towards the NatA substrate peptide SESS 24 in an in vitro [ 14 C]-Ac-CoA–based acetylation assay. Control reactions were performed with no enzyme or no peptide to account for background signal. The immunoprecipitation (IP) and activity measurements were performed in three independent setups, each with three technical replicates per assay. One representative setup is shown. Figure 3—source data 1. Identification of a potential Naa10 homolog.
    Figure Legend Snippet: ( A ) Correlation of Naa10 alteration state on amino-terminal acetylation in mouse embryonic fibroblasts (MEFs). Each dot (n = 533) represents the average amino-terminal acetylation percentage of five replicates of Naa10 +/Y and Naa10 -/Y , respectively. Dashed lines are the borders of ±10% difference. Except for the 10 dots, 522 of the 533 dots are within the borders. The marginal histograms show the distribution of amino-terminal acetylation data points. ( B ) Immunoprecipitation of Naa15. Liver tissue from WT and Naa10 KO mouse was lysed and incubated with anti-Naa15 antibody to retrieve NatA complexes. Proteins were separated by SDS-PAGE and immunoblots probed with anti-Naa15 antibody and anti-NAA10 antibody. ( C ) Catalytic activity of immunoprecipitated NatA. The catalytic activity of NatA precipitated from WT and Naa10 KO mouse liver tissue by anti-Naa15 was measured towards the NatA substrate peptide SESS 24 in an in vitro [ 14 C]-Ac-CoA–based acetylation assay. Control reactions were performed with no enzyme or no peptide to account for background signal. The immunoprecipitation (IP) and activity measurements were performed in three independent setups, each with three technical replicates per assay. One representative setup is shown. Figure 3—source data 1. Identification of a potential Naa10 homolog.

    Techniques Used: Immunoprecipitation, Incubation, SDS Page, Western Blot, Activity Assay, In Vitro, Acetylation Assay

    ( A ) Expression of Naa10 and Naa12 in wildtype (WT), Naa10 KO and Naa12 KO tissues, adult mice 13 weeks of age, (brain, heart, kidney, and testis) by RT-PCR. Expression of GAPDH was analyzed as an endogenous control. ( B ) Phenotypes in Naa12 KO mice. Lack of hypopigmentation (upper; N = 29) and lack of supernumerary ribs (middle and bottom; E18.5; N = 7) in Naa12 KO mouse.
    Figure Legend Snippet: ( A ) Expression of Naa10 and Naa12 in wildtype (WT), Naa10 KO and Naa12 KO tissues, adult mice 13 weeks of age, (brain, heart, kidney, and testis) by RT-PCR. Expression of GAPDH was analyzed as an endogenous control. ( B ) Phenotypes in Naa12 KO mice. Lack of hypopigmentation (upper; N = 29) and lack of supernumerary ribs (middle and bottom; E18.5; N = 7) in Naa12 KO mouse.

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction

    ( A ) In vitro N-terminal acetyltransferase radioactive-based assay. Comparison of mouse Naa10 and Naa12 towards Naa10 peptide substrates, beta-actin (DDDIA-) and gamma-actin (EEEIA-), and the optimal NatA complex peptide substrate, SESSS-. Background control reactions were performed in the absence of either peptide or enzyme. Assays were performed in triplicate; error bars represent SEM. ( B ) Co-immunoprecipitation assay. HEK293 cells were transfected as indicated and lysed after 48 hr. Cell lysates were incubated with 1 μg anti-V5 antibody to precipitate V5-tagged Naa15. The isolated complexes were separated on SDS-PAGE and probed with the indicated antibodies. ( C ) Recombinant mouse Naa12/human Naa15 chimera complex activity. Radioactive acetyltransferase activity assay evaluating the activity of mNaa12-hNaa15 towards peptide (closed circles, ‘mNaa12-hNaa15’) and peptide chemical acetylation in the absence of enzyme (closed circles, ‘Buffer’) as well as chemical acetylation of the enzyme in the absence of peptide (open circles) assay and background (open circles). Error bars represent SD of two technical replicates. These are the same results from fraction #14 (both SESSS- and No Peptide) and both Buffer and Background used to illustrate the size-exclusion-purified mNaa12-hNaa15 complex activity in . Figure 4—source data 1. Characterization of a potential Naa10 homolog.
    Figure Legend Snippet: ( A ) In vitro N-terminal acetyltransferase radioactive-based assay. Comparison of mouse Naa10 and Naa12 towards Naa10 peptide substrates, beta-actin (DDDIA-) and gamma-actin (EEEIA-), and the optimal NatA complex peptide substrate, SESSS-. Background control reactions were performed in the absence of either peptide or enzyme. Assays were performed in triplicate; error bars represent SEM. ( B ) Co-immunoprecipitation assay. HEK293 cells were transfected as indicated and lysed after 48 hr. Cell lysates were incubated with 1 μg anti-V5 antibody to precipitate V5-tagged Naa15. The isolated complexes were separated on SDS-PAGE and probed with the indicated antibodies. ( C ) Recombinant mouse Naa12/human Naa15 chimera complex activity. Radioactive acetyltransferase activity assay evaluating the activity of mNaa12-hNaa15 towards peptide (closed circles, ‘mNaa12-hNaa15’) and peptide chemical acetylation in the absence of enzyme (closed circles, ‘Buffer’) as well as chemical acetylation of the enzyme in the absence of peptide (open circles) assay and background (open circles). Error bars represent SD of two technical replicates. These are the same results from fraction #14 (both SESSS- and No Peptide) and both Buffer and Background used to illustrate the size-exclusion-purified mNaa12-hNaa15 complex activity in . Figure 4—source data 1. Characterization of a potential Naa10 homolog.

    Techniques Used: In Vitro, Radioactivity, Co-Immunoprecipitation Assay, Transfection, Incubation, Isolation, SDS Page, Recombinant, Activity Assay, Purification

     Naa10,  Naa11, and Naa12 peptides identified by LC-MS/MS analysis in Naa15 IP samples from WT and Naa10 -KO mouse.
    Figure Legend Snippet: Naa10, Naa11, and Naa12 peptides identified by LC-MS/MS analysis in Naa15 IP samples from WT and Naa10 -KO mouse.

    Techniques Used: Sequencing

    Naa10 Naa12 DKO exhibit embryonic lethality. Pedigree and genotypes of pups and embryos at E10.5 and E18.5 from Naa10 +/- Naa12 +/- female mice crossed to the Naa10 +/Y Naa12 +/- male mice.
    Figure Legend Snippet: Naa10 Naa12 DKO exhibit embryonic lethality. Pedigree and genotypes of pups and embryos at E10.5 and E18.5 from Naa10 +/- Naa12 +/- female mice crossed to the Naa10 +/Y Naa12 +/- male mice.

    Techniques Used:

    Litter size of  Naa10  × Naa12 matings.
    Figure Legend Snippet: Litter size of Naa10 × Naa12 matings.

    Techniques Used:

    Naa10 Naa12 double-knockout (DKO) exhibit embryonic lethality. Pedigree of mating and genotypes of pups and embryos at E8.5, E10.5, E12.5, and E18.5.
    Figure Legend Snippet: Naa10 Naa12 double-knockout (DKO) exhibit embryonic lethality. Pedigree of mating and genotypes of pups and embryos at E8.5, E10.5, E12.5, and E18.5.

    Techniques Used: Double Knockout

    ( A ) Male body weight for the Naa10 mice on inbred genetic background (eight backcrosses to C57bl6/J). ( B ) Female body weight for the Naa10 mice on inbred genetic background (eight backcrosses to C57bl6/J). ( C ) Male body weight for the Naa10 and Naa12 mice on mixed genetic background. ( D ) Female body weight for the Naa10 and Naa12 mice on mixed genetic background.
    Figure Legend Snippet: ( A ) Male body weight for the Naa10 mice on inbred genetic background (eight backcrosses to C57bl6/J). ( B ) Female body weight for the Naa10 mice on inbred genetic background (eight backcrosses to C57bl6/J). ( C ) Male body weight for the Naa10 and Naa12 mice on mixed genetic background. ( D ) Female body weight for the Naa10 and Naa12 mice on mixed genetic background.

    Techniques Used:

    Model D 4 genotype survival by age.
    Figure Legend Snippet: Model D 4 genotype survival by age.

    Techniques Used:


    Figure Legend Snippet:

    Techniques Used: Isolation, Sequencing, Software

    rabbit monoclonal anti naa10  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit monoclonal anti naa10
    (A) Schematic illustration of the Naa10tm1a mice. (B) PCR confirmation of <t>Naa10</t> deficiency. (C) Confirmation of Naa10 protein in kidney tissue by Western blot. Naa10 protein is not detected in Naa10 -/Y mouse. (D) Expression pattern of Naa10 in the embryo. β-gal staining represents Naa10 localization.
    Rabbit Monoclonal Anti Naa10, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit monoclonal anti naa10/product/Cell Signaling Technology Inc
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit monoclonal anti naa10 - by Bioz Stars, 2023-02
    93/100 stars

    Images

    1) Product Images from "Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway"

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    Journal: bioRxiv

    doi: 10.1101/2020.12.19.422860

    (A) Schematic illustration of the Naa10tm1a mice. (B) PCR confirmation of Naa10 deficiency. (C) Confirmation of Naa10 protein in kidney tissue by Western blot. Naa10 protein is not detected in Naa10 -/Y mouse. (D) Expression pattern of Naa10 in the embryo. β-gal staining represents Naa10 localization.
    Figure Legend Snippet: (A) Schematic illustration of the Naa10tm1a mice. (B) PCR confirmation of Naa10 deficiency. (C) Confirmation of Naa10 protein in kidney tissue by Western blot. Naa10 protein is not detected in Naa10 -/Y mouse. (D) Expression pattern of Naa10 in the embryo. β-gal staining represents Naa10 localization.

    Techniques Used: Western Blot, Expressing, Staining

    (A) Naa10 +/Y , Naa10 -/Y and Naa10 tm1a/Y embryos at E10.5. Growth retardation (5/33, more than 5 somites lower or undersized compared to littermate controls), kinky trunk and developmental arrest are shown in Naa10 -/Y (4/33) and Naa10 tm1a/Y (1/5). Scale bars: 500μm. (B) The percentage lethality in newborns, comparing Naa10 wild-type ( Naa10 +/Y and Naa10 +/+ ), Naa10 -/+ and Naa10 -/Y pups until P3, derived from matings between heterozygous females and wild-type (WT) males. Approximately 11.6% (10/86) of WT, 24% (13/54) of Naa10 +/- and 76.3% (29/38) Naa10 -/Y mice were found dead before P3. (C) Representative images of Naa10 -/Y pups during early postnatal days compared with Naa10 +/Y . Severe developmental defects such as malformations of head and lower body (one leg; black arrowheads), whole- body edema and anophthalmia (black arrows) are shown (N=1 each). (D) Hematoxylin and Eosin (H&E)-stained heart transverse section at E14.5 and vertical section at E18.5, comparing Naa10 +/Y and Naa10 -/Y embryos. Naa10 -/Y embryo shows a VSD at E14.5 and E18.5. Also, at E18.5, Naa10 -/Y embryo shows ASD. Arrow indicates VSD, ASD and DORV. Scale bars: 20 μm. VSD, ventricular septal defect; ASD, atrial septal defect; DORV, double outlet right ventricle.
    Figure Legend Snippet: (A) Naa10 +/Y , Naa10 -/Y and Naa10 tm1a/Y embryos at E10.5. Growth retardation (5/33, more than 5 somites lower or undersized compared to littermate controls), kinky trunk and developmental arrest are shown in Naa10 -/Y (4/33) and Naa10 tm1a/Y (1/5). Scale bars: 500μm. (B) The percentage lethality in newborns, comparing Naa10 wild-type ( Naa10 +/Y and Naa10 +/+ ), Naa10 -/+ and Naa10 -/Y pups until P3, derived from matings between heterozygous females and wild-type (WT) males. Approximately 11.6% (10/86) of WT, 24% (13/54) of Naa10 +/- and 76.3% (29/38) Naa10 -/Y mice were found dead before P3. (C) Representative images of Naa10 -/Y pups during early postnatal days compared with Naa10 +/Y . Severe developmental defects such as malformations of head and lower body (one leg; black arrowheads), whole- body edema and anophthalmia (black arrows) are shown (N=1 each). (D) Hematoxylin and Eosin (H&E)-stained heart transverse section at E14.5 and vertical section at E18.5, comparing Naa10 +/Y and Naa10 -/Y embryos. Naa10 -/Y embryo shows a VSD at E14.5 and E18.5. Also, at E18.5, Naa10 -/Y embryo shows ASD. Arrow indicates VSD, ASD and DORV. Scale bars: 20 μm. VSD, ventricular septal defect; ASD, atrial septal defect; DORV, double outlet right ventricle.

    Techniques Used: Derivative Assay, Staining


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    (A) Wildtype male heart outflow tract region indicating separate aorta and pulmonary trunks nestled between left and right atria. (B) Naa10 -/- female heart from dying P0 pup only has a single outflow tract emerging from the right ventricle, resulting in persistent truncus arteriosus. (C) Naa10 -/y male heart from dying P0 pup has separate outflow tracts but both emerge from the right ventricle, resulting in double outlet right ventricle. (D) normal histology from wildtype male heart of pulmonary artery exiting the right ventricle. (E) histology from Naa10 -/- heart of single outflow tract exiting the right ventricle with tricuspid valve leaflets. (F) histology from Naa10 -/y heart of both pulmonary and aortic arteries emerging from right ventricle within the same plane. (G) Naa10 -/- histology showing membranous VSD, (H) Naa10 -/y histology showing muscular VSD. (I) normal heart revealing closed ductus arterious, (J) Naa10 -/- histology showing open ductus arterious leading to pulmonary overload and likely lethality.
    Figure Legend Snippet: (A) Wildtype male heart outflow tract region indicating separate aorta and pulmonary trunks nestled between left and right atria. (B) Naa10 -/- female heart from dying P0 pup only has a single outflow tract emerging from the right ventricle, resulting in persistent truncus arteriosus. (C) Naa10 -/y male heart from dying P0 pup has separate outflow tracts but both emerge from the right ventricle, resulting in double outlet right ventricle. (D) normal histology from wildtype male heart of pulmonary artery exiting the right ventricle. (E) histology from Naa10 -/- heart of single outflow tract exiting the right ventricle with tricuspid valve leaflets. (F) histology from Naa10 -/y heart of both pulmonary and aortic arteries emerging from right ventricle within the same plane. (G) Naa10 -/- histology showing membranous VSD, (H) Naa10 -/y histology showing muscular VSD. (I) normal heart revealing closed ductus arterious, (J) Naa10 -/- histology showing open ductus arterious leading to pulmonary overload and likely lethality.

    Techniques Used:

    (A-C) Representative images of abnormalities in Naa10 -/Y compared with Naa10 +/Y . (A) Body weight of male (left) and female (right) verses ages was monitored from 2 weeks. The weight of Naa10 -/Y and Naa10 -/- mice is markedly reduced compared with that of the WT mice. Asterisks indicate a statistical difference calculated by Student’s t -test: *p < 0.05. (B) Representative images of completely penetrant phenotypes. Hypopigmentation ( Naa10 +/Y , n=243; Naa10 -/Y , n=121; Naa10 tm1a/Y , n=17) and supernumerary ribs ( Naa10 +/Y , n=3; Naa10 -/Y , n=6; Naa10 tm1a/Y , n=2; E18.5) were found 100% in Naa10 deficient mice. (C) Naa10 -/Y is smaller in size and has round-shaped head ( Naa10 +/Y 0/59, Naa10 -/Y 7/33). Over time, hydrocephaly became apparent. (N=14/29 (∼48%) for >P7 male Naa10 -Y ; N=7/19 (∼36%) for >P7 female Naa10 -/- ). Hydronephrosis (red arrow, Naa10 +/Y 0/23, Naa10 -/Y 14/29, Naa10 +/+ 0/5, Naa10 -/- 7/19) and abnormal genitalia (black arrow) of male (middle, Naa10 +/Y 0/23, Naa10 -/Y 16/29) and female (bottom, hydrometrocolpos, Naa10 +/+ 0/5, Naa10 -/- 7/19) are shown.
    Figure Legend Snippet: (A-C) Representative images of abnormalities in Naa10 -/Y compared with Naa10 +/Y . (A) Body weight of male (left) and female (right) verses ages was monitored from 2 weeks. The weight of Naa10 -/Y and Naa10 -/- mice is markedly reduced compared with that of the WT mice. Asterisks indicate a statistical difference calculated by Student’s t -test: *p < 0.05. (B) Representative images of completely penetrant phenotypes. Hypopigmentation ( Naa10 +/Y , n=243; Naa10 -/Y , n=121; Naa10 tm1a/Y , n=17) and supernumerary ribs ( Naa10 +/Y , n=3; Naa10 -/Y , n=6; Naa10 tm1a/Y , n=2; E18.5) were found 100% in Naa10 deficient mice. (C) Naa10 -/Y is smaller in size and has round-shaped head ( Naa10 +/Y 0/59, Naa10 -/Y 7/33). Over time, hydrocephaly became apparent. (N=14/29 (∼48%) for >P7 male Naa10 -Y ; N=7/19 (∼36%) for >P7 female Naa10 -/- ). Hydronephrosis (red arrow, Naa10 +/Y 0/23, Naa10 -/Y 14/29, Naa10 +/+ 0/5, Naa10 -/- 7/19) and abnormal genitalia (black arrow) of male (middle, Naa10 +/Y 0/23, Naa10 -/Y 16/29) and female (bottom, hydrometrocolpos, Naa10 +/+ 0/5, Naa10 -/- 7/19) are shown.

    Techniques Used:

    (A) In WT mice 13 thoracic vertebrae and ribs are numbered whereas 14 thoracic vertebrae and ribs are counted in mutants ( Naa10 -/Y ) (WT on the left, mutant on the right). n=11 CT scans for Naa10 -/Y compared to n=18 Naa10 +/Y . (B-D) Different number of ribs are linking the sternum between in Naa10 -/Y , Naa10 -/- and WT. (B) 7 ribs linking the sternum in WT. (C) 8 ribs linking the sternum (the white arrow shows the 8th rib) in Naa10 -/Y . (D) 7 on one side + 7 and one almost linking on the other side. In 2 mice, an asymmetrical link was observed. White arrow shows the eighth asymmetrical rib. (E and F) Abnormalities in the cervical phenotype. (E) Cervical WT/ morphology. (F) Partial fusion of C1 and C2 dorsal arch in one mutant mouse.
    Figure Legend Snippet: (A) In WT mice 13 thoracic vertebrae and ribs are numbered whereas 14 thoracic vertebrae and ribs are counted in mutants ( Naa10 -/Y ) (WT on the left, mutant on the right). n=11 CT scans for Naa10 -/Y compared to n=18 Naa10 +/Y . (B-D) Different number of ribs are linking the sternum between in Naa10 -/Y , Naa10 -/- and WT. (B) 7 ribs linking the sternum in WT. (C) 8 ribs linking the sternum (the white arrow shows the 8th rib) in Naa10 -/Y . (D) 7 on one side + 7 and one almost linking on the other side. In 2 mice, an asymmetrical link was observed. White arrow shows the eighth asymmetrical rib. (E and F) Abnormalities in the cervical phenotype. (E) Cervical WT/ morphology. (F) Partial fusion of C1 and C2 dorsal arch in one mutant mouse.

    Techniques Used: Mutagenesis


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    (A) Representative images and histology of renal defects at E18.5 (n=6 out of 39 examined) and P3 (n= 4 out of 11 examined). (B) Hydrocephaly (n=3 CT scans for Naa10 -/Y mice with hydrocephaly compared to n=3 Naa10 -/Y mice without hydrocephaly). (C) Kaplan-Meier survival curve of the Naa10 +/Y and Naa10 -/Y male mice starting at 4 days of life, thus not including any mice that died in the first 3 days of life.
    Figure Legend Snippet: (A) Representative images and histology of renal defects at E18.5 (n=6 out of 39 examined) and P3 (n= 4 out of 11 examined). (B) Hydrocephaly (n=3 CT scans for Naa10 -/Y mice with hydrocephaly compared to n=3 Naa10 -/Y mice without hydrocephaly). (C) Kaplan-Meier survival curve of the Naa10 +/Y and Naa10 -/Y male mice starting at 4 days of life, thus not including any mice that died in the first 3 days of life.

    Techniques Used:

    (A) Correlation of Naa10 alteration state on amino-terminal acetylation in mouse embryonic fibroblasts (MEFs). Each dot (n=533) represents the average amino-terminal acetylation percentage of 5 replicates of Naa10 +/Y and Naa10 -/Y , respectively. Dashed lines are the borders of ±10% difference. Except for the ten dots, 522 of the 533 dots are within the borders. The marginal histograms show the distribution of amino- terminal acetylation data points. (B) Immunoprecipitation of Naa15. Liver tissue from WT and Naa10 KO mouse was lysed and incubated with anti-Naa15 antibody to retrieve NatA complexes. Proteins were separated by SDS-PAGE and immunoblots probed with anti-Naa15 antibody and anti-NAA10 antibody. (C) Catalytic activity of immunoprecipitated NatA. The catalytic activity of NatA precipitated from WT and Naa10 KO mouse liver tissue by anti-Naa15 was measured towards the NatA substrate peptide SESS 24 in an in vitro [ 14 C]-Ac-CoA–Based Acetylation Assay. Control reactions were performed with no enzyme or no peptide to account for background signal. The immunoprecipitation (IP) and activity measurements were performed in three independent setups, each with three technical replicates per assay. One representative setup is shown.
    Figure Legend Snippet: (A) Correlation of Naa10 alteration state on amino-terminal acetylation in mouse embryonic fibroblasts (MEFs). Each dot (n=533) represents the average amino-terminal acetylation percentage of 5 replicates of Naa10 +/Y and Naa10 -/Y , respectively. Dashed lines are the borders of ±10% difference. Except for the ten dots, 522 of the 533 dots are within the borders. The marginal histograms show the distribution of amino- terminal acetylation data points. (B) Immunoprecipitation of Naa15. Liver tissue from WT and Naa10 KO mouse was lysed and incubated with anti-Naa15 antibody to retrieve NatA complexes. Proteins were separated by SDS-PAGE and immunoblots probed with anti-Naa15 antibody and anti-NAA10 antibody. (C) Catalytic activity of immunoprecipitated NatA. The catalytic activity of NatA precipitated from WT and Naa10 KO mouse liver tissue by anti-Naa15 was measured towards the NatA substrate peptide SESS 24 in an in vitro [ 14 C]-Ac-CoA–Based Acetylation Assay. Control reactions were performed with no enzyme or no peptide to account for background signal. The immunoprecipitation (IP) and activity measurements were performed in three independent setups, each with three technical replicates per assay. One representative setup is shown.

    Techniques Used: Immunoprecipitation, Incubation, SDS Page, Western Blot, Activity Assay, In Vitro, Acetylation Assay

    (A) Construction of Naa10 Southern blot probe. (B) Southern blot membrane after hybridization with a Naa10 probe. Expected size band, restricted with Apa I and Xba I, were showed. (C) Ribosome profiling traces for the potential Naa10 paralog (Gm16286, UniProt: Q9CQX6). Picture was modified from GWIPS genome browser, Chr 18, 80206601-80212942.
    Figure Legend Snippet: (A) Construction of Naa10 Southern blot probe. (B) Southern blot membrane after hybridization with a Naa10 probe. Expected size band, restricted with Apa I and Xba I, were showed. (C) Ribosome profiling traces for the potential Naa10 paralog (Gm16286, UniProt: Q9CQX6). Picture was modified from GWIPS genome browser, Chr 18, 80206601-80212942.

    Techniques Used: Southern Blot, Hybridization, Modification

    (A) qPCR analyses of mouse NATs (mNATs) in Naa10 WT and KO adult mouse tissue. (B) Sequence alignment of mNaa10 isoforms and paralogs, including the potential Naa10 paralog mNaa12 (Gm16286, UniProt: Q9CQX6) using Clustal Omega (EMBL-EBI). The peptide used for immunization of rabbits to generate a specific antibody is indicated in red. (C) Full length mNAT cDNA from mouse was amplified and cloned into pGEX-4T1. Proteins were expressed in E. coli BL21 (DE3) and purified via GSH-Sepharose. Shown is a Coomassie stain of fraction 1-4. (D) Cross-reactivity and sensitivity of the used NAT antibodies. 1-20 pmol of GST-mNAT proteins were separated on SDS- PAGE followed by western blot, probed with the indicated antibodies. (E) Recombinant mouse Naa12/human Naa15 chimera complex. Silver-stained denaturing SDS-PAGE (left) containing fractions (#10-20) eluted from an S200 size-exclusion (SEC) chromatography column with fractions evaluated for NatA-type activity indicated (asterisks, below gel) with (F) corresponding radioactive acetyltransferase activity assay comparing the activity of indicated fractions and buffer control (chemical acetylation) towards the SESSS- peptide (filled circles), in the absence of peptide (open circles), and assay background (x). Error bars represent SD of two technical replicates.
    Figure Legend Snippet: (A) qPCR analyses of mouse NATs (mNATs) in Naa10 WT and KO adult mouse tissue. (B) Sequence alignment of mNaa10 isoforms and paralogs, including the potential Naa10 paralog mNaa12 (Gm16286, UniProt: Q9CQX6) using Clustal Omega (EMBL-EBI). The peptide used for immunization of rabbits to generate a specific antibody is indicated in red. (C) Full length mNAT cDNA from mouse was amplified and cloned into pGEX-4T1. Proteins were expressed in E. coli BL21 (DE3) and purified via GSH-Sepharose. Shown is a Coomassie stain of fraction 1-4. (D) Cross-reactivity and sensitivity of the used NAT antibodies. 1-20 pmol of GST-mNAT proteins were separated on SDS- PAGE followed by western blot, probed with the indicated antibodies. (E) Recombinant mouse Naa12/human Naa15 chimera complex. Silver-stained denaturing SDS-PAGE (left) containing fractions (#10-20) eluted from an S200 size-exclusion (SEC) chromatography column with fractions evaluated for NatA-type activity indicated (asterisks, below gel) with (F) corresponding radioactive acetyltransferase activity assay comparing the activity of indicated fractions and buffer control (chemical acetylation) towards the SESSS- peptide (filled circles), in the absence of peptide (open circles), and assay background (x). Error bars represent SD of two technical replicates.

    Techniques Used: Sequencing, Amplification, Clone Assay, Purification, Staining, SDS Page, Western Blot, Recombinant, Chromatography, Activity Assay

    (A) Expression of Naa10 and Naa12 in WT, Naa10 KO and Naa12 KO tissues, adult mice 13 weeks of age, (brain, heart, kidney and testis) by RT-PCR. Expression of GAPDH was analyzed as an endogenous control. (B) Phenotypes in Naa12 KO mice. Lack of hypopigmentation (upper; N=29) and lack of supernumerary ribs (middle and bottom; E18.5; N=7) in Naa12 KO mouse.
    Figure Legend Snippet: (A) Expression of Naa10 and Naa12 in WT, Naa10 KO and Naa12 KO tissues, adult mice 13 weeks of age, (brain, heart, kidney and testis) by RT-PCR. Expression of GAPDH was analyzed as an endogenous control. (B) Phenotypes in Naa12 KO mice. Lack of hypopigmentation (upper; N=29) and lack of supernumerary ribs (middle and bottom; E18.5; N=7) in Naa12 KO mouse.

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction

    (A) In vitro N-terminal acetyltransferase radioactive-based assay. Comparison of mouse Naa10 and Naa12 towards Naa10 peptide substrates, beta-actin (DDDIA-) and gamma-actin (EEEIA-), and the optimal NatA complex peptide substrate, SESSS-. Background control reactions were performed in the absence of either peptide or enzyme. Assays were performed in triplicate; error bars represent S.E.M. (B) Co-immunoprecipitation assay. HEK293 cells were transfected as indicated and lysed after 48 h. Cell lysates were incubated with 1 μg anti-V5 antibody to precipitate V5-tagged Naa15. The isolated complexes were separated on SDS-PAGE and probed with the indicated antibodies. (C) Recombinant mouse Naa12/human Naa15 chimera complex activity. Radioactive acetyltransferase activity assay evaluating the activity of mNaa12-hNaa15 towards peptide (closed circles, “mNaa12-hNaa15”) and peptide chemical acetylation in the absence of enzyme (closed circles, “Buffer”) as well as chemical acetylation of the the enzyme in the absence of peptide (open circles) assay and background (open circles). Error bars represent SD of two technical replicates. These are the same results from fraction #14 (both SESSS- and No Peptide) and both Buffer and Background used to illustrate the size-exclusion-purified mNaa12-hNaa15 complex activity in Supplement Fig. 6F.
    Figure Legend Snippet: (A) In vitro N-terminal acetyltransferase radioactive-based assay. Comparison of mouse Naa10 and Naa12 towards Naa10 peptide substrates, beta-actin (DDDIA-) and gamma-actin (EEEIA-), and the optimal NatA complex peptide substrate, SESSS-. Background control reactions were performed in the absence of either peptide or enzyme. Assays were performed in triplicate; error bars represent S.E.M. (B) Co-immunoprecipitation assay. HEK293 cells were transfected as indicated and lysed after 48 h. Cell lysates were incubated with 1 μg anti-V5 antibody to precipitate V5-tagged Naa15. The isolated complexes were separated on SDS-PAGE and probed with the indicated antibodies. (C) Recombinant mouse Naa12/human Naa15 chimera complex activity. Radioactive acetyltransferase activity assay evaluating the activity of mNaa12-hNaa15 towards peptide (closed circles, “mNaa12-hNaa15”) and peptide chemical acetylation in the absence of enzyme (closed circles, “Buffer”) as well as chemical acetylation of the the enzyme in the absence of peptide (open circles) assay and background (open circles). Error bars represent SD of two technical replicates. These are the same results from fraction #14 (both SESSS- and No Peptide) and both Buffer and Background used to illustrate the size-exclusion-purified mNaa12-hNaa15 complex activity in Supplement Fig. 6F.

    Techniques Used: In Vitro, Radioactivity, Co-Immunoprecipitation Assay, Transfection, Incubation, Isolation, SDS Page, Recombinant, Activity Assay, Purification


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    Naa10 Naa12 DKO exhibit embryonic lethality. Pedigree and genotypes of pups and embryos at E10.5 and E18.5 from Naa10 +/- Naa12 +/- female mice crossed to the Naa10 +/Y Naa12 +/- male mice.
    Figure Legend Snippet: Naa10 Naa12 DKO exhibit embryonic lethality. Pedigree and genotypes of pups and embryos at E10.5 and E18.5 from Naa10 +/- Naa12 +/- female mice crossed to the Naa10 +/Y Naa12 +/- male mice.

    Techniques Used:

    Naa10 Naa12 DKO exhibit embryonic lethality. Pedigree of mating and genotypes of pups and embryos at E8.5, E10.5, E12.5 and E18.5.
    Figure Legend Snippet: Naa10 Naa12 DKO exhibit embryonic lethality. Pedigree of mating and genotypes of pups and embryos at E8.5, E10.5, E12.5 and E18.5.

    Techniques Used:

    (A) Male body weight for the Naa10 mice on inbred genetic background (8 backcrosses to C57bl6/J). (B) Female body weight for the Naa10 mice on inbred genetic background (8 backcrosses to C57bl6/J). (C) Male body weight for the Naa10 and Naa12 mice on mixed genetic background. (D) Female body weight for the Naa10 and Naa12 mice on mixed genetic background.
    Figure Legend Snippet: (A) Male body weight for the Naa10 mice on inbred genetic background (8 backcrosses to C57bl6/J). (B) Female body weight for the Naa10 mice on inbred genetic background (8 backcrosses to C57bl6/J). (C) Male body weight for the Naa10 and Naa12 mice on mixed genetic background. (D) Female body weight for the Naa10 and Naa12 mice on mixed genetic background.

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    rabbit monoclonal anti naa10  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
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    Cell Signaling Technology Inc rabbit monoclonal anti naa10
    (A) Schematic illustration of the Naa10tm1a mice. (B) PCR confirmation of <t>Naa10</t> deficiency. (C) Confirmation of Naa10 protein in kidney tissue by Western blot. Naa10 protein is not detected in Naa10 -/Y mouse. (D) Expression pattern of Naa10 in the embryo. β-gal staining represents Naa10 localization.
    Rabbit Monoclonal Anti Naa10, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit monoclonal anti naa10/product/Cell Signaling Technology Inc
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit monoclonal anti naa10 - by Bioz Stars, 2023-02
    93/100 stars

    Images

    1) Product Images from "Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway"

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    Journal: bioRxiv

    doi: 10.1101/2020.12.19.422860

    (A) Schematic illustration of the Naa10tm1a mice. (B) PCR confirmation of Naa10 deficiency. (C) Confirmation of Naa10 protein in kidney tissue by Western blot. Naa10 protein is not detected in Naa10 -/Y mouse. (D) Expression pattern of Naa10 in the embryo. β-gal staining represents Naa10 localization.
    Figure Legend Snippet: (A) Schematic illustration of the Naa10tm1a mice. (B) PCR confirmation of Naa10 deficiency. (C) Confirmation of Naa10 protein in kidney tissue by Western blot. Naa10 protein is not detected in Naa10 -/Y mouse. (D) Expression pattern of Naa10 in the embryo. β-gal staining represents Naa10 localization.

    Techniques Used: Western Blot, Expressing, Staining

    (A) Naa10 +/Y , Naa10 -/Y and Naa10 tm1a/Y embryos at E10.5. Growth retardation (5/33, more than 5 somites lower or undersized compared to littermate controls), kinky trunk and developmental arrest are shown in Naa10 -/Y (4/33) and Naa10 tm1a/Y (1/5). Scale bars: 500μm. (B) The percentage lethality in newborns, comparing Naa10 wild-type ( Naa10 +/Y and Naa10 +/+ ), Naa10 -/+ and Naa10 -/Y pups until P3, derived from matings between heterozygous females and wild-type (WT) males. Approximately 11.6% (10/86) of WT, 24% (13/54) of Naa10 +/- and 76.3% (29/38) Naa10 -/Y mice were found dead before P3. (C) Representative images of Naa10 -/Y pups during early postnatal days compared with Naa10 +/Y . Severe developmental defects such as malformations of head and lower body (one leg; black arrowheads), whole- body edema and anophthalmia (black arrows) are shown (N=1 each). (D) Hematoxylin and Eosin (H&E)-stained heart transverse section at E14.5 and vertical section at E18.5, comparing Naa10 +/Y and Naa10 -/Y embryos. Naa10 -/Y embryo shows a VSD at E14.5 and E18.5. Also, at E18.5, Naa10 -/Y embryo shows ASD. Arrow indicates VSD, ASD and DORV. Scale bars: 20 μm. VSD, ventricular septal defect; ASD, atrial septal defect; DORV, double outlet right ventricle.
    Figure Legend Snippet: (A) Naa10 +/Y , Naa10 -/Y and Naa10 tm1a/Y embryos at E10.5. Growth retardation (5/33, more than 5 somites lower or undersized compared to littermate controls), kinky trunk and developmental arrest are shown in Naa10 -/Y (4/33) and Naa10 tm1a/Y (1/5). Scale bars: 500μm. (B) The percentage lethality in newborns, comparing Naa10 wild-type ( Naa10 +/Y and Naa10 +/+ ), Naa10 -/+ and Naa10 -/Y pups until P3, derived from matings between heterozygous females and wild-type (WT) males. Approximately 11.6% (10/86) of WT, 24% (13/54) of Naa10 +/- and 76.3% (29/38) Naa10 -/Y mice were found dead before P3. (C) Representative images of Naa10 -/Y pups during early postnatal days compared with Naa10 +/Y . Severe developmental defects such as malformations of head and lower body (one leg; black arrowheads), whole- body edema and anophthalmia (black arrows) are shown (N=1 each). (D) Hematoxylin and Eosin (H&E)-stained heart transverse section at E14.5 and vertical section at E18.5, comparing Naa10 +/Y and Naa10 -/Y embryos. Naa10 -/Y embryo shows a VSD at E14.5 and E18.5. Also, at E18.5, Naa10 -/Y embryo shows ASD. Arrow indicates VSD, ASD and DORV. Scale bars: 20 μm. VSD, ventricular septal defect; ASD, atrial septal defect; DORV, double outlet right ventricle.

    Techniques Used: Derivative Assay, Staining


    Figure Legend Snippet:

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    Figure Legend Snippet:

    Techniques Used:

    (A) Wildtype male heart outflow tract region indicating separate aorta and pulmonary trunks nestled between left and right atria. (B) Naa10 -/- female heart from dying P0 pup only has a single outflow tract emerging from the right ventricle, resulting in persistent truncus arteriosus. (C) Naa10 -/y male heart from dying P0 pup has separate outflow tracts but both emerge from the right ventricle, resulting in double outlet right ventricle. (D) normal histology from wildtype male heart of pulmonary artery exiting the right ventricle. (E) histology from Naa10 -/- heart of single outflow tract exiting the right ventricle with tricuspid valve leaflets. (F) histology from Naa10 -/y heart of both pulmonary and aortic arteries emerging from right ventricle within the same plane. (G) Naa10 -/- histology showing membranous VSD, (H) Naa10 -/y histology showing muscular VSD. (I) normal heart revealing closed ductus arterious, (J) Naa10 -/- histology showing open ductus arterious leading to pulmonary overload and likely lethality.
    Figure Legend Snippet: (A) Wildtype male heart outflow tract region indicating separate aorta and pulmonary trunks nestled between left and right atria. (B) Naa10 -/- female heart from dying P0 pup only has a single outflow tract emerging from the right ventricle, resulting in persistent truncus arteriosus. (C) Naa10 -/y male heart from dying P0 pup has separate outflow tracts but both emerge from the right ventricle, resulting in double outlet right ventricle. (D) normal histology from wildtype male heart of pulmonary artery exiting the right ventricle. (E) histology from Naa10 -/- heart of single outflow tract exiting the right ventricle with tricuspid valve leaflets. (F) histology from Naa10 -/y heart of both pulmonary and aortic arteries emerging from right ventricle within the same plane. (G) Naa10 -/- histology showing membranous VSD, (H) Naa10 -/y histology showing muscular VSD. (I) normal heart revealing closed ductus arterious, (J) Naa10 -/- histology showing open ductus arterious leading to pulmonary overload and likely lethality.

    Techniques Used:

    (A-C) Representative images of abnormalities in Naa10 -/Y compared with Naa10 +/Y . (A) Body weight of male (left) and female (right) verses ages was monitored from 2 weeks. The weight of Naa10 -/Y and Naa10 -/- mice is markedly reduced compared with that of the WT mice. Asterisks indicate a statistical difference calculated by Student’s t -test: *p < 0.05. (B) Representative images of completely penetrant phenotypes. Hypopigmentation ( Naa10 +/Y , n=243; Naa10 -/Y , n=121; Naa10 tm1a/Y , n=17) and supernumerary ribs ( Naa10 +/Y , n=3; Naa10 -/Y , n=6; Naa10 tm1a/Y , n=2; E18.5) were found 100% in Naa10 deficient mice. (C) Naa10 -/Y is smaller in size and has round-shaped head ( Naa10 +/Y 0/59, Naa10 -/Y 7/33). Over time, hydrocephaly became apparent. (N=14/29 (∼48%) for >P7 male Naa10 -Y ; N=7/19 (∼36%) for >P7 female Naa10 -/- ). Hydronephrosis (red arrow, Naa10 +/Y 0/23, Naa10 -/Y 14/29, Naa10 +/+ 0/5, Naa10 -/- 7/19) and abnormal genitalia (black arrow) of male (middle, Naa10 +/Y 0/23, Naa10 -/Y 16/29) and female (bottom, hydrometrocolpos, Naa10 +/+ 0/5, Naa10 -/- 7/19) are shown.
    Figure Legend Snippet: (A-C) Representative images of abnormalities in Naa10 -/Y compared with Naa10 +/Y . (A) Body weight of male (left) and female (right) verses ages was monitored from 2 weeks. The weight of Naa10 -/Y and Naa10 -/- mice is markedly reduced compared with that of the WT mice. Asterisks indicate a statistical difference calculated by Student’s t -test: *p < 0.05. (B) Representative images of completely penetrant phenotypes. Hypopigmentation ( Naa10 +/Y , n=243; Naa10 -/Y , n=121; Naa10 tm1a/Y , n=17) and supernumerary ribs ( Naa10 +/Y , n=3; Naa10 -/Y , n=6; Naa10 tm1a/Y , n=2; E18.5) were found 100% in Naa10 deficient mice. (C) Naa10 -/Y is smaller in size and has round-shaped head ( Naa10 +/Y 0/59, Naa10 -/Y 7/33). Over time, hydrocephaly became apparent. (N=14/29 (∼48%) for >P7 male Naa10 -Y ; N=7/19 (∼36%) for >P7 female Naa10 -/- ). Hydronephrosis (red arrow, Naa10 +/Y 0/23, Naa10 -/Y 14/29, Naa10 +/+ 0/5, Naa10 -/- 7/19) and abnormal genitalia (black arrow) of male (middle, Naa10 +/Y 0/23, Naa10 -/Y 16/29) and female (bottom, hydrometrocolpos, Naa10 +/+ 0/5, Naa10 -/- 7/19) are shown.

    Techniques Used:

    (A) In WT mice 13 thoracic vertebrae and ribs are numbered whereas 14 thoracic vertebrae and ribs are counted in mutants ( Naa10 -/Y ) (WT on the left, mutant on the right). n=11 CT scans for Naa10 -/Y compared to n=18 Naa10 +/Y . (B-D) Different number of ribs are linking the sternum between in Naa10 -/Y , Naa10 -/- and WT. (B) 7 ribs linking the sternum in WT. (C) 8 ribs linking the sternum (the white arrow shows the 8th rib) in Naa10 -/Y . (D) 7 on one side + 7 and one almost linking on the other side. In 2 mice, an asymmetrical link was observed. White arrow shows the eighth asymmetrical rib. (E and F) Abnormalities in the cervical phenotype. (E) Cervical WT/ morphology. (F) Partial fusion of C1 and C2 dorsal arch in one mutant mouse.
    Figure Legend Snippet: (A) In WT mice 13 thoracic vertebrae and ribs are numbered whereas 14 thoracic vertebrae and ribs are counted in mutants ( Naa10 -/Y ) (WT on the left, mutant on the right). n=11 CT scans for Naa10 -/Y compared to n=18 Naa10 +/Y . (B-D) Different number of ribs are linking the sternum between in Naa10 -/Y , Naa10 -/- and WT. (B) 7 ribs linking the sternum in WT. (C) 8 ribs linking the sternum (the white arrow shows the 8th rib) in Naa10 -/Y . (D) 7 on one side + 7 and one almost linking on the other side. In 2 mice, an asymmetrical link was observed. White arrow shows the eighth asymmetrical rib. (E and F) Abnormalities in the cervical phenotype. (E) Cervical WT/ morphology. (F) Partial fusion of C1 and C2 dorsal arch in one mutant mouse.

    Techniques Used: Mutagenesis


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    (A) Representative images and histology of renal defects at E18.5 (n=6 out of 39 examined) and P3 (n= 4 out of 11 examined). (B) Hydrocephaly (n=3 CT scans for Naa10 -/Y mice with hydrocephaly compared to n=3 Naa10 -/Y mice without hydrocephaly). (C) Kaplan-Meier survival curve of the Naa10 +/Y and Naa10 -/Y male mice starting at 4 days of life, thus not including any mice that died in the first 3 days of life.
    Figure Legend Snippet: (A) Representative images and histology of renal defects at E18.5 (n=6 out of 39 examined) and P3 (n= 4 out of 11 examined). (B) Hydrocephaly (n=3 CT scans for Naa10 -/Y mice with hydrocephaly compared to n=3 Naa10 -/Y mice without hydrocephaly). (C) Kaplan-Meier survival curve of the Naa10 +/Y and Naa10 -/Y male mice starting at 4 days of life, thus not including any mice that died in the first 3 days of life.

    Techniques Used:

    (A) Correlation of Naa10 alteration state on amino-terminal acetylation in mouse embryonic fibroblasts (MEFs). Each dot (n=533) represents the average amino-terminal acetylation percentage of 5 replicates of Naa10 +/Y and Naa10 -/Y , respectively. Dashed lines are the borders of ±10% difference. Except for the ten dots, 522 of the 533 dots are within the borders. The marginal histograms show the distribution of amino- terminal acetylation data points. (B) Immunoprecipitation of Naa15. Liver tissue from WT and Naa10 KO mouse was lysed and incubated with anti-Naa15 antibody to retrieve NatA complexes. Proteins were separated by SDS-PAGE and immunoblots probed with anti-Naa15 antibody and anti-NAA10 antibody. (C) Catalytic activity of immunoprecipitated NatA. The catalytic activity of NatA precipitated from WT and Naa10 KO mouse liver tissue by anti-Naa15 was measured towards the NatA substrate peptide SESS 24 in an in vitro [ 14 C]-Ac-CoA–Based Acetylation Assay. Control reactions were performed with no enzyme or no peptide to account for background signal. The immunoprecipitation (IP) and activity measurements were performed in three independent setups, each with three technical replicates per assay. One representative setup is shown.
    Figure Legend Snippet: (A) Correlation of Naa10 alteration state on amino-terminal acetylation in mouse embryonic fibroblasts (MEFs). Each dot (n=533) represents the average amino-terminal acetylation percentage of 5 replicates of Naa10 +/Y and Naa10 -/Y , respectively. Dashed lines are the borders of ±10% difference. Except for the ten dots, 522 of the 533 dots are within the borders. The marginal histograms show the distribution of amino- terminal acetylation data points. (B) Immunoprecipitation of Naa15. Liver tissue from WT and Naa10 KO mouse was lysed and incubated with anti-Naa15 antibody to retrieve NatA complexes. Proteins were separated by SDS-PAGE and immunoblots probed with anti-Naa15 antibody and anti-NAA10 antibody. (C) Catalytic activity of immunoprecipitated NatA. The catalytic activity of NatA precipitated from WT and Naa10 KO mouse liver tissue by anti-Naa15 was measured towards the NatA substrate peptide SESS 24 in an in vitro [ 14 C]-Ac-CoA–Based Acetylation Assay. Control reactions were performed with no enzyme or no peptide to account for background signal. The immunoprecipitation (IP) and activity measurements were performed in three independent setups, each with three technical replicates per assay. One representative setup is shown.

    Techniques Used: Immunoprecipitation, Incubation, SDS Page, Western Blot, Activity Assay, In Vitro, Acetylation Assay

    (A) Construction of Naa10 Southern blot probe. (B) Southern blot membrane after hybridization with a Naa10 probe. Expected size band, restricted with Apa I and Xba I, were showed. (C) Ribosome profiling traces for the potential Naa10 paralog (Gm16286, UniProt: Q9CQX6). Picture was modified from GWIPS genome browser, Chr 18, 80206601-80212942.
    Figure Legend Snippet: (A) Construction of Naa10 Southern blot probe. (B) Southern blot membrane after hybridization with a Naa10 probe. Expected size band, restricted with Apa I and Xba I, were showed. (C) Ribosome profiling traces for the potential Naa10 paralog (Gm16286, UniProt: Q9CQX6). Picture was modified from GWIPS genome browser, Chr 18, 80206601-80212942.

    Techniques Used: Southern Blot, Hybridization, Modification

    (A) qPCR analyses of mouse NATs (mNATs) in Naa10 WT and KO adult mouse tissue. (B) Sequence alignment of mNaa10 isoforms and paralogs, including the potential Naa10 paralog mNaa12 (Gm16286, UniProt: Q9CQX6) using Clustal Omega (EMBL-EBI). The peptide used for immunization of rabbits to generate a specific antibody is indicated in red. (C) Full length mNAT cDNA from mouse was amplified and cloned into pGEX-4T1. Proteins were expressed in E. coli BL21 (DE3) and purified via GSH-Sepharose. Shown is a Coomassie stain of fraction 1-4. (D) Cross-reactivity and sensitivity of the used NAT antibodies. 1-20 pmol of GST-mNAT proteins were separated on SDS- PAGE followed by western blot, probed with the indicated antibodies. (E) Recombinant mouse Naa12/human Naa15 chimera complex. Silver-stained denaturing SDS-PAGE (left) containing fractions (#10-20) eluted from an S200 size-exclusion (SEC) chromatography column with fractions evaluated for NatA-type activity indicated (asterisks, below gel) with (F) corresponding radioactive acetyltransferase activity assay comparing the activity of indicated fractions and buffer control (chemical acetylation) towards the SESSS- peptide (filled circles), in the absence of peptide (open circles), and assay background (x). Error bars represent SD of two technical replicates.
    Figure Legend Snippet: (A) qPCR analyses of mouse NATs (mNATs) in Naa10 WT and KO adult mouse tissue. (B) Sequence alignment of mNaa10 isoforms and paralogs, including the potential Naa10 paralog mNaa12 (Gm16286, UniProt: Q9CQX6) using Clustal Omega (EMBL-EBI). The peptide used for immunization of rabbits to generate a specific antibody is indicated in red. (C) Full length mNAT cDNA from mouse was amplified and cloned into pGEX-4T1. Proteins were expressed in E. coli BL21 (DE3) and purified via GSH-Sepharose. Shown is a Coomassie stain of fraction 1-4. (D) Cross-reactivity and sensitivity of the used NAT antibodies. 1-20 pmol of GST-mNAT proteins were separated on SDS- PAGE followed by western blot, probed with the indicated antibodies. (E) Recombinant mouse Naa12/human Naa15 chimera complex. Silver-stained denaturing SDS-PAGE (left) containing fractions (#10-20) eluted from an S200 size-exclusion (SEC) chromatography column with fractions evaluated for NatA-type activity indicated (asterisks, below gel) with (F) corresponding radioactive acetyltransferase activity assay comparing the activity of indicated fractions and buffer control (chemical acetylation) towards the SESSS- peptide (filled circles), in the absence of peptide (open circles), and assay background (x). Error bars represent SD of two technical replicates.

    Techniques Used: Sequencing, Amplification, Clone Assay, Purification, Staining, SDS Page, Western Blot, Recombinant, Chromatography, Activity Assay

    (A) Expression of Naa10 and Naa12 in WT, Naa10 KO and Naa12 KO tissues, adult mice 13 weeks of age, (brain, heart, kidney and testis) by RT-PCR. Expression of GAPDH was analyzed as an endogenous control. (B) Phenotypes in Naa12 KO mice. Lack of hypopigmentation (upper; N=29) and lack of supernumerary ribs (middle and bottom; E18.5; N=7) in Naa12 KO mouse.
    Figure Legend Snippet: (A) Expression of Naa10 and Naa12 in WT, Naa10 KO and Naa12 KO tissues, adult mice 13 weeks of age, (brain, heart, kidney and testis) by RT-PCR. Expression of GAPDH was analyzed as an endogenous control. (B) Phenotypes in Naa12 KO mice. Lack of hypopigmentation (upper; N=29) and lack of supernumerary ribs (middle and bottom; E18.5; N=7) in Naa12 KO mouse.

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction

    (A) In vitro N-terminal acetyltransferase radioactive-based assay. Comparison of mouse Naa10 and Naa12 towards Naa10 peptide substrates, beta-actin (DDDIA-) and gamma-actin (EEEIA-), and the optimal NatA complex peptide substrate, SESSS-. Background control reactions were performed in the absence of either peptide or enzyme. Assays were performed in triplicate; error bars represent S.E.M. (B) Co-immunoprecipitation assay. HEK293 cells were transfected as indicated and lysed after 48 h. Cell lysates were incubated with 1 μg anti-V5 antibody to precipitate V5-tagged Naa15. The isolated complexes were separated on SDS-PAGE and probed with the indicated antibodies. (C) Recombinant mouse Naa12/human Naa15 chimera complex activity. Radioactive acetyltransferase activity assay evaluating the activity of mNaa12-hNaa15 towards peptide (closed circles, “mNaa12-hNaa15”) and peptide chemical acetylation in the absence of enzyme (closed circles, “Buffer”) as well as chemical acetylation of the the enzyme in the absence of peptide (open circles) assay and background (open circles). Error bars represent SD of two technical replicates. These are the same results from fraction #14 (both SESSS- and No Peptide) and both Buffer and Background used to illustrate the size-exclusion-purified mNaa12-hNaa15 complex activity in Supplement Fig. 6F.
    Figure Legend Snippet: (A) In vitro N-terminal acetyltransferase radioactive-based assay. Comparison of mouse Naa10 and Naa12 towards Naa10 peptide substrates, beta-actin (DDDIA-) and gamma-actin (EEEIA-), and the optimal NatA complex peptide substrate, SESSS-. Background control reactions were performed in the absence of either peptide or enzyme. Assays were performed in triplicate; error bars represent S.E.M. (B) Co-immunoprecipitation assay. HEK293 cells were transfected as indicated and lysed after 48 h. Cell lysates were incubated with 1 μg anti-V5 antibody to precipitate V5-tagged Naa15. The isolated complexes were separated on SDS-PAGE and probed with the indicated antibodies. (C) Recombinant mouse Naa12/human Naa15 chimera complex activity. Radioactive acetyltransferase activity assay evaluating the activity of mNaa12-hNaa15 towards peptide (closed circles, “mNaa12-hNaa15”) and peptide chemical acetylation in the absence of enzyme (closed circles, “Buffer”) as well as chemical acetylation of the the enzyme in the absence of peptide (open circles) assay and background (open circles). Error bars represent SD of two technical replicates. These are the same results from fraction #14 (both SESSS- and No Peptide) and both Buffer and Background used to illustrate the size-exclusion-purified mNaa12-hNaa15 complex activity in Supplement Fig. 6F.

    Techniques Used: In Vitro, Radioactivity, Co-Immunoprecipitation Assay, Transfection, Incubation, Isolation, SDS Page, Recombinant, Activity Assay, Purification


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    Naa10 Naa12 DKO exhibit embryonic lethality. Pedigree and genotypes of pups and embryos at E10.5 and E18.5 from Naa10 +/- Naa12 +/- female mice crossed to the Naa10 +/Y Naa12 +/- male mice.
    Figure Legend Snippet: Naa10 Naa12 DKO exhibit embryonic lethality. Pedigree and genotypes of pups and embryos at E10.5 and E18.5 from Naa10 +/- Naa12 +/- female mice crossed to the Naa10 +/Y Naa12 +/- male mice.

    Techniques Used:

    Naa10 Naa12 DKO exhibit embryonic lethality. Pedigree of mating and genotypes of pups and embryos at E8.5, E10.5, E12.5 and E18.5.
    Figure Legend Snippet: Naa10 Naa12 DKO exhibit embryonic lethality. Pedigree of mating and genotypes of pups and embryos at E8.5, E10.5, E12.5 and E18.5.

    Techniques Used:

    (A) Male body weight for the Naa10 mice on inbred genetic background (8 backcrosses to C57bl6/J). (B) Female body weight for the Naa10 mice on inbred genetic background (8 backcrosses to C57bl6/J). (C) Male body weight for the Naa10 and Naa12 mice on mixed genetic background. (D) Female body weight for the Naa10 and Naa12 mice on mixed genetic background.
    Figure Legend Snippet: (A) Male body weight for the Naa10 mice on inbred genetic background (8 backcrosses to C57bl6/J). (B) Female body weight for the Naa10 mice on inbred genetic background (8 backcrosses to C57bl6/J). (C) Male body weight for the Naa10 and Naa12 mice on mixed genetic background. (D) Female body weight for the Naa10 and Naa12 mice on mixed genetic background.

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    naa10 rabbit monoclonal antibody  (Cell Signaling Technology Inc)


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    Structured Review

    Cell Signaling Technology Inc naa10 rabbit monoclonal antibody
    a hNAA50(magenta), hNAA15(green), and hNAA10(orange) within the hNatE complex overlay with ScNatE (gray, PDB: 6O07). The α3 helix and β7 strand of Naa50 is shown to shift toward <t>Naa10</t> in the human structure ( b ) hNatE aligned with hNatA (light blue, PDB:6C9M) and hNAA50 (wheat, PDB: 3TFY). The top zoom-in area shows the alignment of free hNAA50 and hNatE. The below zoom-in area shows the hNAA10 conformational change induced by hNAA50 binding.
    Naa10 Rabbit Monoclonal Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/naa10 rabbit monoclonal antibody/product/Cell Signaling Technology Inc
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    Images

    1) Product Images from "Molecular basis for N-terminal acetylation by human NatE and its modulation by HYPK"

    Article Title: Molecular basis for N-terminal acetylation by human NatE and its modulation by HYPK

    Journal: Nature Communications

    doi: 10.1038/s41467-020-14584-7

    a hNAA50(magenta), hNAA15(green), and hNAA10(orange) within the hNatE complex overlay with ScNatE (gray, PDB: 6O07). The α3 helix and β7 strand of Naa50 is shown to shift toward Naa10 in the human structure ( b ) hNatE aligned with hNatA (light blue, PDB:6C9M) and hNAA50 (wheat, PDB: 3TFY). The top zoom-in area shows the alignment of free hNAA50 and hNatE. The below zoom-in area shows the hNAA10 conformational change induced by hNAA50 binding.
    Figure Legend Snippet: a hNAA50(magenta), hNAA15(green), and hNAA10(orange) within the hNatE complex overlay with ScNatE (gray, PDB: 6O07). The α3 helix and β7 strand of Naa50 is shown to shift toward Naa10 in the human structure ( b ) hNatE aligned with hNatA (light blue, PDB:6C9M) and hNAA50 (wheat, PDB: 3TFY). The top zoom-in area shows the alignment of free hNAA50 and hNatE. The below zoom-in area shows the hNAA10 conformational change induced by hNAA50 binding.

    Techniques Used: Binding Assay

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    Cell Signaling Technology Inc rabbit monoclonal anti naa10
    ( A ) <t>Naa10</t> +/Y , Naa10 -/Y , and Naa10 tm1a/Y embryos at E10.5. Growth retardation (5/33, more than five somites lower or undersized compared to littermate controls), kinky trunk, and developmental arrest are shown in Naa10 -/Y (4/33) and Naa10 tm1a/Y (1/5). Scale bars: 500 μm. ( B ) The percentage lethality in newborns, comparing Naa10 wildtype (WT) ( Naa10 +/Y and Naa10 +/+ ), Naa10 -/+ and Naa10 -/Y pups until P3, derived from matings between heterozygous females and WT males. Approximately 11.6% (10/86) of WT, 24% (13/54) of Naa10 +/- , and 76.3% (29/38) of Naa10 -/Y mice were found dead before P3. ( C ) Representative images of Naa10 -/Y pups during early postnatal days compared with Naa10 +/Y . Severe developmental defects such as malformations of head and lower body (one leg; black arrowheads), whole-body edema, and anophthalmia (black arrows) are shown (N = 1 each). ( D ) Hematoxylin and eosin (H&E)-stained heart transverse section at E14.5 and vertical section at E18.5, comparing Naa10 +/Y and Naa10 -/Y embryos. Naa10 -/Y embryo shows a ventricular septal defect (VSD) at E14.5 and E18.5. Also, at E18.5, Naa10 -/Y embryo shows atrial septal defect (ASD). Arrow indicates VSD, ASD, and double outlet right ventricle (DORV). Scale bars: 20 μm.
    Rabbit Monoclonal Anti Naa10, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit monoclonal anti naa10/product/Cell Signaling Technology Inc
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    Cell Signaling Technology Inc anti naa10
    ( A ) <t>Naa10</t> +/Y , Naa10 -/Y , and Naa10 tm1a/Y embryos at E10.5. Growth retardation (5/33, more than five somites lower or undersized compared to littermate controls), kinky trunk, and developmental arrest are shown in Naa10 -/Y (4/33) and Naa10 tm1a/Y (1/5). Scale bars: 500 μm. ( B ) The percentage lethality in newborns, comparing Naa10 wildtype (WT) ( Naa10 +/Y and Naa10 +/+ ), Naa10 -/+ and Naa10 -/Y pups until P3, derived from matings between heterozygous females and WT males. Approximately 11.6% (10/86) of WT, 24% (13/54) of Naa10 +/- , and 76.3% (29/38) of Naa10 -/Y mice were found dead before P3. ( C ) Representative images of Naa10 -/Y pups during early postnatal days compared with Naa10 +/Y . Severe developmental defects such as malformations of head and lower body (one leg; black arrowheads), whole-body edema, and anophthalmia (black arrows) are shown (N = 1 each). ( D ) Hematoxylin and eosin (H&E)-stained heart transverse section at E14.5 and vertical section at E18.5, comparing Naa10 +/Y and Naa10 -/Y embryos. Naa10 -/Y embryo shows a ventricular septal defect (VSD) at E14.5 and E18.5. Also, at E18.5, Naa10 -/Y embryo shows atrial septal defect (ASD). Arrow indicates VSD, ASD, and double outlet right ventricle (DORV). Scale bars: 20 μm.
    Anti Naa10, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti naa10/product/Cell Signaling Technology Inc
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti naa10 - by Bioz Stars, 2023-02
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    93
    Cell Signaling Technology Inc naa10 rabbit monoclonal antibody
    a hNAA50(magenta), hNAA15(green), and hNAA10(orange) within the hNatE complex overlay with ScNatE (gray, PDB: 6O07). The α3 helix and β7 strand of Naa50 is shown to shift toward <t>Naa10</t> in the human structure ( b ) hNatE aligned with hNatA (light blue, PDB:6C9M) and hNAA50 (wheat, PDB: 3TFY). The top zoom-in area shows the alignment of free hNAA50 and hNatE. The below zoom-in area shows the hNAA10 conformational change induced by hNAA50 binding.
    Naa10 Rabbit Monoclonal Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    ( A ) Naa10 +/Y , Naa10 -/Y , and Naa10 tm1a/Y embryos at E10.5. Growth retardation (5/33, more than five somites lower or undersized compared to littermate controls), kinky trunk, and developmental arrest are shown in Naa10 -/Y (4/33) and Naa10 tm1a/Y (1/5). Scale bars: 500 μm. ( B ) The percentage lethality in newborns, comparing Naa10 wildtype (WT) ( Naa10 +/Y and Naa10 +/+ ), Naa10 -/+ and Naa10 -/Y pups until P3, derived from matings between heterozygous females and WT males. Approximately 11.6% (10/86) of WT, 24% (13/54) of Naa10 +/- , and 76.3% (29/38) of Naa10 -/Y mice were found dead before P3. ( C ) Representative images of Naa10 -/Y pups during early postnatal days compared with Naa10 +/Y . Severe developmental defects such as malformations of head and lower body (one leg; black arrowheads), whole-body edema, and anophthalmia (black arrows) are shown (N = 1 each). ( D ) Hematoxylin and eosin (H&E)-stained heart transverse section at E14.5 and vertical section at E18.5, comparing Naa10 +/Y and Naa10 -/Y embryos. Naa10 -/Y embryo shows a ventricular septal defect (VSD) at E14.5 and E18.5. Also, at E18.5, Naa10 -/Y embryo shows atrial septal defect (ASD). Arrow indicates VSD, ASD, and double outlet right ventricle (DORV). Scale bars: 20 μm.

    Journal: eLife

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    doi: 10.7554/eLife.65952

    Figure Lengend Snippet: ( A ) Naa10 +/Y , Naa10 -/Y , and Naa10 tm1a/Y embryos at E10.5. Growth retardation (5/33, more than five somites lower or undersized compared to littermate controls), kinky trunk, and developmental arrest are shown in Naa10 -/Y (4/33) and Naa10 tm1a/Y (1/5). Scale bars: 500 μm. ( B ) The percentage lethality in newborns, comparing Naa10 wildtype (WT) ( Naa10 +/Y and Naa10 +/+ ), Naa10 -/+ and Naa10 -/Y pups until P3, derived from matings between heterozygous females and WT males. Approximately 11.6% (10/86) of WT, 24% (13/54) of Naa10 +/- , and 76.3% (29/38) of Naa10 -/Y mice were found dead before P3. ( C ) Representative images of Naa10 -/Y pups during early postnatal days compared with Naa10 +/Y . Severe developmental defects such as malformations of head and lower body (one leg; black arrowheads), whole-body edema, and anophthalmia (black arrows) are shown (N = 1 each). ( D ) Hematoxylin and eosin (H&E)-stained heart transverse section at E14.5 and vertical section at E18.5, comparing Naa10 +/Y and Naa10 -/Y embryos. Naa10 -/Y embryo shows a ventricular septal defect (VSD) at E14.5 and E18.5. Also, at E18.5, Naa10 -/Y embryo shows atrial septal defect (ASD). Arrow indicates VSD, ASD, and double outlet right ventricle (DORV). Scale bars: 20 μm.

    Article Snippet: The following antibodies were used: rabbit anti-Naa10 (Abcam #ab155687), rabbit anti-Naa10 (Protein Tech #14803-1-AP), rabbit monoclonal anti-NAA10 (Cell Signaling, #13357), goat anti-Naa10 (Santa Cruz, #sc-33256), rabbit anti-Naa10 (Santa Cruz, #sc-33820), rabbit anti-Naa11 (Novus Biologicals; #NBP1-90853), mouse anti-Naa15/NARG1 (Abcam; #ab60065), rabbit polyclonal anti-NAA15 , rabbit anti-Naa50 (LifeSpan BioSciences; #LS-C81324-100), rabbit anti-FLAG (Sigma; #F7425), mouse anti-GAPDH (Abcam; #ab9484), goat anti-actin (Santa Cruz, #1615), mouse anti-GST (GenScript; #A00865), and mouse anti-V5 (Life Technologies; #R960-25).

    Techniques: Derivative Assay, Staining

    ( A ) Wildtype male heart outflow tract region indicating separate aorta and pulmonary trunks nestled between left and right atria. ( B ) Naa10 -/- female heart from dying P0 pup only has a single outflow tract emerging from the right ventricle, resulting in persistent truncus arteriosus. ( C ) Naa10 -/y male heart from dying P0 pup has separate outflow tracts but both emerge from the right ventricle, resulting in double outlet right ventricle. ( D ) Normal histology from wildtype male heart of pulmonary artery exiting the right ventricle. ( E ) Histology from Naa10 -/- heart of single outflow tract exiting the right ventricle with tricuspid valve leaflets. ( F ) Histology from Naa10 -/y heart of both pulmonary and aortic arteries emerging from right ventricle within the same plane. ( G ) Naa10 -/- histology showing membranous ventricular septal defect (VSD). ( H ) Naa10 -/y histology showing muscular VSD. ( I ) Normal heart revealing closed ductus arterious. ( J ) Naa10 -/- histology showing open ductus arterious leading to pulmonary overload and likely lethality.

    Journal: eLife

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    doi: 10.7554/eLife.65952

    Figure Lengend Snippet: ( A ) Wildtype male heart outflow tract region indicating separate aorta and pulmonary trunks nestled between left and right atria. ( B ) Naa10 -/- female heart from dying P0 pup only has a single outflow tract emerging from the right ventricle, resulting in persistent truncus arteriosus. ( C ) Naa10 -/y male heart from dying P0 pup has separate outflow tracts but both emerge from the right ventricle, resulting in double outlet right ventricle. ( D ) Normal histology from wildtype male heart of pulmonary artery exiting the right ventricle. ( E ) Histology from Naa10 -/- heart of single outflow tract exiting the right ventricle with tricuspid valve leaflets. ( F ) Histology from Naa10 -/y heart of both pulmonary and aortic arteries emerging from right ventricle within the same plane. ( G ) Naa10 -/- histology showing membranous ventricular septal defect (VSD). ( H ) Naa10 -/y histology showing muscular VSD. ( I ) Normal heart revealing closed ductus arterious. ( J ) Naa10 -/- histology showing open ductus arterious leading to pulmonary overload and likely lethality.

    Article Snippet: The following antibodies were used: rabbit anti-Naa10 (Abcam #ab155687), rabbit anti-Naa10 (Protein Tech #14803-1-AP), rabbit monoclonal anti-NAA10 (Cell Signaling, #13357), goat anti-Naa10 (Santa Cruz, #sc-33256), rabbit anti-Naa10 (Santa Cruz, #sc-33820), rabbit anti-Naa11 (Novus Biologicals; #NBP1-90853), mouse anti-Naa15/NARG1 (Abcam; #ab60065), rabbit polyclonal anti-NAA15 , rabbit anti-Naa50 (LifeSpan BioSciences; #LS-C81324-100), rabbit anti-FLAG (Sigma; #F7425), mouse anti-GAPDH (Abcam; #ab9484), goat anti-actin (Santa Cruz, #1615), mouse anti-GST (GenScript; #A00865), and mouse anti-V5 (Life Technologies; #R960-25).

    Techniques:

    ( A–C ) Representative images of abnormalities in Naa10 -/Y compared with Naa10 +/Y . ( A ) Body weight of male (left) and female (right) versus ages was monitored from 2 weeks. The weight of Naa10 -/Y and Naa10 -/- mice is markedly reduced compared with that of the wildtype (WT) mice. Asterisks indicate a statistical difference calculated by Student’s t -test: *p<0.05. ( B ) Representative images of completely penetrant phenotypes. Hypopigmentation ( Naa10 +/Y , n = 243; Naa10 -/Y , n = 121; Naa10 tm1a/Y , n = 17) and supernumerary ribs ( Naa10 +/Y , n = 3; Naa10 -/Y , n = 6; Naa10 tm1a/Y , n = 2; E18.5) were found 100% in Naa10 -deficient mice. ( C ) Naa10 -/Y is smaller in size and has round-shaped head ( Naa10 +/Y 0/59, Naa10 -/Y 7/33). Over time, hydrocephaly became apparent (N = 14/29 [~48%] for >P7 male Naa10 -Y ; N = 7/19 [~36%] for >P7 female Naa10 -/- ). Hydronephrosis (red arrow, Naa10 +/Y 0/23, Naa10 -/Y 14/29, Naa10 +/+ 0/5, Naa10 -/- 7/19) and abnormal genitalia (black arrow) of male (middle, Naa10 +/Y 0/23, Naa10 -/Y 16/29) and female (bottom, hydrometrocolpos, Naa10 +/+ 0/5, Naa10 -/- 7/19) are shown.

    Journal: eLife

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    doi: 10.7554/eLife.65952

    Figure Lengend Snippet: ( A–C ) Representative images of abnormalities in Naa10 -/Y compared with Naa10 +/Y . ( A ) Body weight of male (left) and female (right) versus ages was monitored from 2 weeks. The weight of Naa10 -/Y and Naa10 -/- mice is markedly reduced compared with that of the wildtype (WT) mice. Asterisks indicate a statistical difference calculated by Student’s t -test: *p<0.05. ( B ) Representative images of completely penetrant phenotypes. Hypopigmentation ( Naa10 +/Y , n = 243; Naa10 -/Y , n = 121; Naa10 tm1a/Y , n = 17) and supernumerary ribs ( Naa10 +/Y , n = 3; Naa10 -/Y , n = 6; Naa10 tm1a/Y , n = 2; E18.5) were found 100% in Naa10 -deficient mice. ( C ) Naa10 -/Y is smaller in size and has round-shaped head ( Naa10 +/Y 0/59, Naa10 -/Y 7/33). Over time, hydrocephaly became apparent (N = 14/29 [~48%] for >P7 male Naa10 -Y ; N = 7/19 [~36%] for >P7 female Naa10 -/- ). Hydronephrosis (red arrow, Naa10 +/Y 0/23, Naa10 -/Y 14/29, Naa10 +/+ 0/5, Naa10 -/- 7/19) and abnormal genitalia (black arrow) of male (middle, Naa10 +/Y 0/23, Naa10 -/Y 16/29) and female (bottom, hydrometrocolpos, Naa10 +/+ 0/5, Naa10 -/- 7/19) are shown.

    Article Snippet: The following antibodies were used: rabbit anti-Naa10 (Abcam #ab155687), rabbit anti-Naa10 (Protein Tech #14803-1-AP), rabbit monoclonal anti-NAA10 (Cell Signaling, #13357), goat anti-Naa10 (Santa Cruz, #sc-33256), rabbit anti-Naa10 (Santa Cruz, #sc-33820), rabbit anti-Naa11 (Novus Biologicals; #NBP1-90853), mouse anti-Naa15/NARG1 (Abcam; #ab60065), rabbit polyclonal anti-NAA15 , rabbit anti-Naa50 (LifeSpan BioSciences; #LS-C81324-100), rabbit anti-FLAG (Sigma; #F7425), mouse anti-GAPDH (Abcam; #ab9484), goat anti-actin (Santa Cruz, #1615), mouse anti-GST (GenScript; #A00865), and mouse anti-V5 (Life Technologies; #R960-25).

    Techniques:

    Skeletal analyses for ribs, sternebrae, and vertebrae.

    Journal: eLife

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    doi: 10.7554/eLife.65952

    Figure Lengend Snippet: Skeletal analyses for ribs, sternebrae, and vertebrae.

    Article Snippet: The following antibodies were used: rabbit anti-Naa10 (Abcam #ab155687), rabbit anti-Naa10 (Protein Tech #14803-1-AP), rabbit monoclonal anti-NAA10 (Cell Signaling, #13357), goat anti-Naa10 (Santa Cruz, #sc-33256), rabbit anti-Naa10 (Santa Cruz, #sc-33820), rabbit anti-Naa11 (Novus Biologicals; #NBP1-90853), mouse anti-Naa15/NARG1 (Abcam; #ab60065), rabbit polyclonal anti-NAA15 , rabbit anti-Naa50 (LifeSpan BioSciences; #LS-C81324-100), rabbit anti-FLAG (Sigma; #F7425), mouse anti-GAPDH (Abcam; #ab9484), goat anti-actin (Santa Cruz, #1615), mouse anti-GST (GenScript; #A00865), and mouse anti-V5 (Life Technologies; #R960-25).

    Techniques:

    ( A ) In wildtype (WT) mice, 13 thoracic vertebrae and ribs are numbered, whereas 14 thoracic vertebrae and ribs are counted in mutants ( Naa10 -/Y ) (WT on the left, mutant on the right). n = 11 CT scans for Naa10 -/Y compared to n = 18 Naa10 +/Y . ( B–D ) Different number of ribs are linking the sternum between in Naa10 -/Y , Naa10 -/- , and WT. ( B ) Seven ribs linking the sternum in WT. ( C ) Eight ribs linking the sternum (the white arrow shows the eighth rib) in Naa10 -/Y . ( D ) Seven on one side plus seven and one almost linking on the other side. In two mice, an asymmetrical link was observed. White arrow shows the eighth asymmetrical rib. ( E, F ) Abnormalities in the cervical phenotype. ( E ) Cervical WT/morphology. ( F ) Partial fusion of C1 and C2 dorsal arch in one mutant mouse.

    Journal: eLife

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    doi: 10.7554/eLife.65952

    Figure Lengend Snippet: ( A ) In wildtype (WT) mice, 13 thoracic vertebrae and ribs are numbered, whereas 14 thoracic vertebrae and ribs are counted in mutants ( Naa10 -/Y ) (WT on the left, mutant on the right). n = 11 CT scans for Naa10 -/Y compared to n = 18 Naa10 +/Y . ( B–D ) Different number of ribs are linking the sternum between in Naa10 -/Y , Naa10 -/- , and WT. ( B ) Seven ribs linking the sternum in WT. ( C ) Eight ribs linking the sternum (the white arrow shows the eighth rib) in Naa10 -/Y . ( D ) Seven on one side plus seven and one almost linking on the other side. In two mice, an asymmetrical link was observed. White arrow shows the eighth asymmetrical rib. ( E, F ) Abnormalities in the cervical phenotype. ( E ) Cervical WT/morphology. ( F ) Partial fusion of C1 and C2 dorsal arch in one mutant mouse.

    Article Snippet: The following antibodies were used: rabbit anti-Naa10 (Abcam #ab155687), rabbit anti-Naa10 (Protein Tech #14803-1-AP), rabbit monoclonal anti-NAA10 (Cell Signaling, #13357), goat anti-Naa10 (Santa Cruz, #sc-33256), rabbit anti-Naa10 (Santa Cruz, #sc-33820), rabbit anti-Naa11 (Novus Biologicals; #NBP1-90853), mouse anti-Naa15/NARG1 (Abcam; #ab60065), rabbit polyclonal anti-NAA15 , rabbit anti-Naa50 (LifeSpan BioSciences; #LS-C81324-100), rabbit anti-FLAG (Sigma; #F7425), mouse anti-GAPDH (Abcam; #ab9484), goat anti-actin (Santa Cruz, #1615), mouse anti-GST (GenScript; #A00865), and mouse anti-V5 (Life Technologies; #R960-25).

    Techniques: Mutagenesis

    ( A ) Representative images and histology of renal defects at E18.5 (n = 6 out of 39 examined) and P3 (n = 4 out of 11 examined). ( B ) Hydrocephaly (n = 3 CT scans for Naa10 -/Y mice with hydrocephaly compared to n = 3 Naa10 -/Y mice without hydrocephaly). ( C ) Kaplan–Meier survival curve of the Naa10 +/Y and Naa10 -/Y male mice starting at 4 days of life, thus not including any mice that died in the first three days of life.

    Journal: eLife

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    doi: 10.7554/eLife.65952

    Figure Lengend Snippet: ( A ) Representative images and histology of renal defects at E18.5 (n = 6 out of 39 examined) and P3 (n = 4 out of 11 examined). ( B ) Hydrocephaly (n = 3 CT scans for Naa10 -/Y mice with hydrocephaly compared to n = 3 Naa10 -/Y mice without hydrocephaly). ( C ) Kaplan–Meier survival curve of the Naa10 +/Y and Naa10 -/Y male mice starting at 4 days of life, thus not including any mice that died in the first three days of life.

    Article Snippet: The following antibodies were used: rabbit anti-Naa10 (Abcam #ab155687), rabbit anti-Naa10 (Protein Tech #14803-1-AP), rabbit monoclonal anti-NAA10 (Cell Signaling, #13357), goat anti-Naa10 (Santa Cruz, #sc-33256), rabbit anti-Naa10 (Santa Cruz, #sc-33820), rabbit anti-Naa11 (Novus Biologicals; #NBP1-90853), mouse anti-Naa15/NARG1 (Abcam; #ab60065), rabbit polyclonal anti-NAA15 , rabbit anti-Naa50 (LifeSpan BioSciences; #LS-C81324-100), rabbit anti-FLAG (Sigma; #F7425), mouse anti-GAPDH (Abcam; #ab9484), goat anti-actin (Santa Cruz, #1615), mouse anti-GST (GenScript; #A00865), and mouse anti-V5 (Life Technologies; #R960-25).

    Techniques:

    ( A ) Correlation of Naa10 alteration state on amino-terminal acetylation in mouse embryonic fibroblasts (MEFs). Each dot (n = 533) represents the average amino-terminal acetylation percentage of five replicates of Naa10 +/Y and Naa10 -/Y , respectively. Dashed lines are the borders of ±10% difference. Except for the 10 dots, 522 of the 533 dots are within the borders. The marginal histograms show the distribution of amino-terminal acetylation data points. ( B ) Immunoprecipitation of Naa15. Liver tissue from WT and Naa10 KO mouse was lysed and incubated with anti-Naa15 antibody to retrieve NatA complexes. Proteins were separated by SDS-PAGE and immunoblots probed with anti-Naa15 antibody and anti-NAA10 antibody. ( C ) Catalytic activity of immunoprecipitated NatA. The catalytic activity of NatA precipitated from WT and Naa10 KO mouse liver tissue by anti-Naa15 was measured towards the NatA substrate peptide SESS 24 in an in vitro [ 14 C]-Ac-CoA–based acetylation assay. Control reactions were performed with no enzyme or no peptide to account for background signal. The immunoprecipitation (IP) and activity measurements were performed in three independent setups, each with three technical replicates per assay. One representative setup is shown. Figure 3—source data 1. Identification of a potential Naa10 homolog.

    Journal: eLife

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    doi: 10.7554/eLife.65952

    Figure Lengend Snippet: ( A ) Correlation of Naa10 alteration state on amino-terminal acetylation in mouse embryonic fibroblasts (MEFs). Each dot (n = 533) represents the average amino-terminal acetylation percentage of five replicates of Naa10 +/Y and Naa10 -/Y , respectively. Dashed lines are the borders of ±10% difference. Except for the 10 dots, 522 of the 533 dots are within the borders. The marginal histograms show the distribution of amino-terminal acetylation data points. ( B ) Immunoprecipitation of Naa15. Liver tissue from WT and Naa10 KO mouse was lysed and incubated with anti-Naa15 antibody to retrieve NatA complexes. Proteins were separated by SDS-PAGE and immunoblots probed with anti-Naa15 antibody and anti-NAA10 antibody. ( C ) Catalytic activity of immunoprecipitated NatA. The catalytic activity of NatA precipitated from WT and Naa10 KO mouse liver tissue by anti-Naa15 was measured towards the NatA substrate peptide SESS 24 in an in vitro [ 14 C]-Ac-CoA–based acetylation assay. Control reactions were performed with no enzyme or no peptide to account for background signal. The immunoprecipitation (IP) and activity measurements were performed in three independent setups, each with three technical replicates per assay. One representative setup is shown. Figure 3—source data 1. Identification of a potential Naa10 homolog.

    Article Snippet: The following antibodies were used: rabbit anti-Naa10 (Abcam #ab155687), rabbit anti-Naa10 (Protein Tech #14803-1-AP), rabbit monoclonal anti-NAA10 (Cell Signaling, #13357), goat anti-Naa10 (Santa Cruz, #sc-33256), rabbit anti-Naa10 (Santa Cruz, #sc-33820), rabbit anti-Naa11 (Novus Biologicals; #NBP1-90853), mouse anti-Naa15/NARG1 (Abcam; #ab60065), rabbit polyclonal anti-NAA15 , rabbit anti-Naa50 (LifeSpan BioSciences; #LS-C81324-100), rabbit anti-FLAG (Sigma; #F7425), mouse anti-GAPDH (Abcam; #ab9484), goat anti-actin (Santa Cruz, #1615), mouse anti-GST (GenScript; #A00865), and mouse anti-V5 (Life Technologies; #R960-25).

    Techniques: Immunoprecipitation, Incubation, SDS Page, Western Blot, Activity Assay, In Vitro, Acetylation Assay

    ( A ) Expression of Naa10 and Naa12 in wildtype (WT), Naa10 KO and Naa12 KO tissues, adult mice 13 weeks of age, (brain, heart, kidney, and testis) by RT-PCR. Expression of GAPDH was analyzed as an endogenous control. ( B ) Phenotypes in Naa12 KO mice. Lack of hypopigmentation (upper; N = 29) and lack of supernumerary ribs (middle and bottom; E18.5; N = 7) in Naa12 KO mouse.

    Journal: eLife

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    doi: 10.7554/eLife.65952

    Figure Lengend Snippet: ( A ) Expression of Naa10 and Naa12 in wildtype (WT), Naa10 KO and Naa12 KO tissues, adult mice 13 weeks of age, (brain, heart, kidney, and testis) by RT-PCR. Expression of GAPDH was analyzed as an endogenous control. ( B ) Phenotypes in Naa12 KO mice. Lack of hypopigmentation (upper; N = 29) and lack of supernumerary ribs (middle and bottom; E18.5; N = 7) in Naa12 KO mouse.

    Article Snippet: The following antibodies were used: rabbit anti-Naa10 (Abcam #ab155687), rabbit anti-Naa10 (Protein Tech #14803-1-AP), rabbit monoclonal anti-NAA10 (Cell Signaling, #13357), goat anti-Naa10 (Santa Cruz, #sc-33256), rabbit anti-Naa10 (Santa Cruz, #sc-33820), rabbit anti-Naa11 (Novus Biologicals; #NBP1-90853), mouse anti-Naa15/NARG1 (Abcam; #ab60065), rabbit polyclonal anti-NAA15 , rabbit anti-Naa50 (LifeSpan BioSciences; #LS-C81324-100), rabbit anti-FLAG (Sigma; #F7425), mouse anti-GAPDH (Abcam; #ab9484), goat anti-actin (Santa Cruz, #1615), mouse anti-GST (GenScript; #A00865), and mouse anti-V5 (Life Technologies; #R960-25).

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction

    ( A ) In vitro N-terminal acetyltransferase radioactive-based assay. Comparison of mouse Naa10 and Naa12 towards Naa10 peptide substrates, beta-actin (DDDIA-) and gamma-actin (EEEIA-), and the optimal NatA complex peptide substrate, SESSS-. Background control reactions were performed in the absence of either peptide or enzyme. Assays were performed in triplicate; error bars represent SEM. ( B ) Co-immunoprecipitation assay. HEK293 cells were transfected as indicated and lysed after 48 hr. Cell lysates were incubated with 1 μg anti-V5 antibody to precipitate V5-tagged Naa15. The isolated complexes were separated on SDS-PAGE and probed with the indicated antibodies. ( C ) Recombinant mouse Naa12/human Naa15 chimera complex activity. Radioactive acetyltransferase activity assay evaluating the activity of mNaa12-hNaa15 towards peptide (closed circles, ‘mNaa12-hNaa15’) and peptide chemical acetylation in the absence of enzyme (closed circles, ‘Buffer’) as well as chemical acetylation of the enzyme in the absence of peptide (open circles) assay and background (open circles). Error bars represent SD of two technical replicates. These are the same results from fraction #14 (both SESSS- and No Peptide) and both Buffer and Background used to illustrate the size-exclusion-purified mNaa12-hNaa15 complex activity in . Figure 4—source data 1. Characterization of a potential Naa10 homolog.

    Journal: eLife

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    doi: 10.7554/eLife.65952

    Figure Lengend Snippet: ( A ) In vitro N-terminal acetyltransferase radioactive-based assay. Comparison of mouse Naa10 and Naa12 towards Naa10 peptide substrates, beta-actin (DDDIA-) and gamma-actin (EEEIA-), and the optimal NatA complex peptide substrate, SESSS-. Background control reactions were performed in the absence of either peptide or enzyme. Assays were performed in triplicate; error bars represent SEM. ( B ) Co-immunoprecipitation assay. HEK293 cells were transfected as indicated and lysed after 48 hr. Cell lysates were incubated with 1 μg anti-V5 antibody to precipitate V5-tagged Naa15. The isolated complexes were separated on SDS-PAGE and probed with the indicated antibodies. ( C ) Recombinant mouse Naa12/human Naa15 chimera complex activity. Radioactive acetyltransferase activity assay evaluating the activity of mNaa12-hNaa15 towards peptide (closed circles, ‘mNaa12-hNaa15’) and peptide chemical acetylation in the absence of enzyme (closed circles, ‘Buffer’) as well as chemical acetylation of the enzyme in the absence of peptide (open circles) assay and background (open circles). Error bars represent SD of two technical replicates. These are the same results from fraction #14 (both SESSS- and No Peptide) and both Buffer and Background used to illustrate the size-exclusion-purified mNaa12-hNaa15 complex activity in . Figure 4—source data 1. Characterization of a potential Naa10 homolog.

    Article Snippet: The following antibodies were used: rabbit anti-Naa10 (Abcam #ab155687), rabbit anti-Naa10 (Protein Tech #14803-1-AP), rabbit monoclonal anti-NAA10 (Cell Signaling, #13357), goat anti-Naa10 (Santa Cruz, #sc-33256), rabbit anti-Naa10 (Santa Cruz, #sc-33820), rabbit anti-Naa11 (Novus Biologicals; #NBP1-90853), mouse anti-Naa15/NARG1 (Abcam; #ab60065), rabbit polyclonal anti-NAA15 , rabbit anti-Naa50 (LifeSpan BioSciences; #LS-C81324-100), rabbit anti-FLAG (Sigma; #F7425), mouse anti-GAPDH (Abcam; #ab9484), goat anti-actin (Santa Cruz, #1615), mouse anti-GST (GenScript; #A00865), and mouse anti-V5 (Life Technologies; #R960-25).

    Techniques: In Vitro, Radioactivity, Co-Immunoprecipitation Assay, Transfection, Incubation, Isolation, SDS Page, Recombinant, Activity Assay, Purification

     Naa10,  Naa11, and Naa12 peptides identified by LC-MS/MS analysis in Naa15 IP samples from WT and Naa10 -KO mouse.

    Journal: eLife

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    doi: 10.7554/eLife.65952

    Figure Lengend Snippet: Naa10, Naa11, and Naa12 peptides identified by LC-MS/MS analysis in Naa15 IP samples from WT and Naa10 -KO mouse.

    Article Snippet: The following antibodies were used: rabbit anti-Naa10 (Abcam #ab155687), rabbit anti-Naa10 (Protein Tech #14803-1-AP), rabbit monoclonal anti-NAA10 (Cell Signaling, #13357), goat anti-Naa10 (Santa Cruz, #sc-33256), rabbit anti-Naa10 (Santa Cruz, #sc-33820), rabbit anti-Naa11 (Novus Biologicals; #NBP1-90853), mouse anti-Naa15/NARG1 (Abcam; #ab60065), rabbit polyclonal anti-NAA15 , rabbit anti-Naa50 (LifeSpan BioSciences; #LS-C81324-100), rabbit anti-FLAG (Sigma; #F7425), mouse anti-GAPDH (Abcam; #ab9484), goat anti-actin (Santa Cruz, #1615), mouse anti-GST (GenScript; #A00865), and mouse anti-V5 (Life Technologies; #R960-25).

    Techniques: Sequencing

    Naa10 Naa12 DKO exhibit embryonic lethality. Pedigree and genotypes of pups and embryos at E10.5 and E18.5 from Naa10 +/- Naa12 +/- female mice crossed to the Naa10 +/Y Naa12 +/- male mice.

    Journal: eLife

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    doi: 10.7554/eLife.65952

    Figure Lengend Snippet: Naa10 Naa12 DKO exhibit embryonic lethality. Pedigree and genotypes of pups and embryos at E10.5 and E18.5 from Naa10 +/- Naa12 +/- female mice crossed to the Naa10 +/Y Naa12 +/- male mice.

    Article Snippet: The following antibodies were used: rabbit anti-Naa10 (Abcam #ab155687), rabbit anti-Naa10 (Protein Tech #14803-1-AP), rabbit monoclonal anti-NAA10 (Cell Signaling, #13357), goat anti-Naa10 (Santa Cruz, #sc-33256), rabbit anti-Naa10 (Santa Cruz, #sc-33820), rabbit anti-Naa11 (Novus Biologicals; #NBP1-90853), mouse anti-Naa15/NARG1 (Abcam; #ab60065), rabbit polyclonal anti-NAA15 , rabbit anti-Naa50 (LifeSpan BioSciences; #LS-C81324-100), rabbit anti-FLAG (Sigma; #F7425), mouse anti-GAPDH (Abcam; #ab9484), goat anti-actin (Santa Cruz, #1615), mouse anti-GST (GenScript; #A00865), and mouse anti-V5 (Life Technologies; #R960-25).

    Techniques:

    Litter size of  Naa10  × Naa12 matings.

    Journal: eLife

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    doi: 10.7554/eLife.65952

    Figure Lengend Snippet: Litter size of Naa10 × Naa12 matings.

    Article Snippet: The following antibodies were used: rabbit anti-Naa10 (Abcam #ab155687), rabbit anti-Naa10 (Protein Tech #14803-1-AP), rabbit monoclonal anti-NAA10 (Cell Signaling, #13357), goat anti-Naa10 (Santa Cruz, #sc-33256), rabbit anti-Naa10 (Santa Cruz, #sc-33820), rabbit anti-Naa11 (Novus Biologicals; #NBP1-90853), mouse anti-Naa15/NARG1 (Abcam; #ab60065), rabbit polyclonal anti-NAA15 , rabbit anti-Naa50 (LifeSpan BioSciences; #LS-C81324-100), rabbit anti-FLAG (Sigma; #F7425), mouse anti-GAPDH (Abcam; #ab9484), goat anti-actin (Santa Cruz, #1615), mouse anti-GST (GenScript; #A00865), and mouse anti-V5 (Life Technologies; #R960-25).

    Techniques:

    Naa10 Naa12 double-knockout (DKO) exhibit embryonic lethality. Pedigree of mating and genotypes of pups and embryos at E8.5, E10.5, E12.5, and E18.5.

    Journal: eLife

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    doi: 10.7554/eLife.65952

    Figure Lengend Snippet: Naa10 Naa12 double-knockout (DKO) exhibit embryonic lethality. Pedigree of mating and genotypes of pups and embryos at E8.5, E10.5, E12.5, and E18.5.

    Article Snippet: The following antibodies were used: rabbit anti-Naa10 (Abcam #ab155687), rabbit anti-Naa10 (Protein Tech #14803-1-AP), rabbit monoclonal anti-NAA10 (Cell Signaling, #13357), goat anti-Naa10 (Santa Cruz, #sc-33256), rabbit anti-Naa10 (Santa Cruz, #sc-33820), rabbit anti-Naa11 (Novus Biologicals; #NBP1-90853), mouse anti-Naa15/NARG1 (Abcam; #ab60065), rabbit polyclonal anti-NAA15 , rabbit anti-Naa50 (LifeSpan BioSciences; #LS-C81324-100), rabbit anti-FLAG (Sigma; #F7425), mouse anti-GAPDH (Abcam; #ab9484), goat anti-actin (Santa Cruz, #1615), mouse anti-GST (GenScript; #A00865), and mouse anti-V5 (Life Technologies; #R960-25).

    Techniques: Double Knockout

    ( A ) Male body weight for the Naa10 mice on inbred genetic background (eight backcrosses to C57bl6/J). ( B ) Female body weight for the Naa10 mice on inbred genetic background (eight backcrosses to C57bl6/J). ( C ) Male body weight for the Naa10 and Naa12 mice on mixed genetic background. ( D ) Female body weight for the Naa10 and Naa12 mice on mixed genetic background.

    Journal: eLife

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    doi: 10.7554/eLife.65952

    Figure Lengend Snippet: ( A ) Male body weight for the Naa10 mice on inbred genetic background (eight backcrosses to C57bl6/J). ( B ) Female body weight for the Naa10 mice on inbred genetic background (eight backcrosses to C57bl6/J). ( C ) Male body weight for the Naa10 and Naa12 mice on mixed genetic background. ( D ) Female body weight for the Naa10 and Naa12 mice on mixed genetic background.

    Article Snippet: The following antibodies were used: rabbit anti-Naa10 (Abcam #ab155687), rabbit anti-Naa10 (Protein Tech #14803-1-AP), rabbit monoclonal anti-NAA10 (Cell Signaling, #13357), goat anti-Naa10 (Santa Cruz, #sc-33256), rabbit anti-Naa10 (Santa Cruz, #sc-33820), rabbit anti-Naa11 (Novus Biologicals; #NBP1-90853), mouse anti-Naa15/NARG1 (Abcam; #ab60065), rabbit polyclonal anti-NAA15 , rabbit anti-Naa50 (LifeSpan BioSciences; #LS-C81324-100), rabbit anti-FLAG (Sigma; #F7425), mouse anti-GAPDH (Abcam; #ab9484), goat anti-actin (Santa Cruz, #1615), mouse anti-GST (GenScript; #A00865), and mouse anti-V5 (Life Technologies; #R960-25).

    Techniques:

    Model D 4 genotype survival by age.

    Journal: eLife

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    doi: 10.7554/eLife.65952

    Figure Lengend Snippet: Model D 4 genotype survival by age.

    Article Snippet: The following antibodies were used: rabbit anti-Naa10 (Abcam #ab155687), rabbit anti-Naa10 (Protein Tech #14803-1-AP), rabbit monoclonal anti-NAA10 (Cell Signaling, #13357), goat anti-Naa10 (Santa Cruz, #sc-33256), rabbit anti-Naa10 (Santa Cruz, #sc-33820), rabbit anti-Naa11 (Novus Biologicals; #NBP1-90853), mouse anti-Naa15/NARG1 (Abcam; #ab60065), rabbit polyclonal anti-NAA15 , rabbit anti-Naa50 (LifeSpan BioSciences; #LS-C81324-100), rabbit anti-FLAG (Sigma; #F7425), mouse anti-GAPDH (Abcam; #ab9484), goat anti-actin (Santa Cruz, #1615), mouse anti-GST (GenScript; #A00865), and mouse anti-V5 (Life Technologies; #R960-25).

    Techniques:

    Journal: eLife

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    doi: 10.7554/eLife.65952

    Figure Lengend Snippet:

    Article Snippet: The following antibodies were used: rabbit anti-Naa10 (Abcam #ab155687), rabbit anti-Naa10 (Protein Tech #14803-1-AP), rabbit monoclonal anti-NAA10 (Cell Signaling, #13357), goat anti-Naa10 (Santa Cruz, #sc-33256), rabbit anti-Naa10 (Santa Cruz, #sc-33820), rabbit anti-Naa11 (Novus Biologicals; #NBP1-90853), mouse anti-Naa15/NARG1 (Abcam; #ab60065), rabbit polyclonal anti-NAA15 , rabbit anti-Naa50 (LifeSpan BioSciences; #LS-C81324-100), rabbit anti-FLAG (Sigma; #F7425), mouse anti-GAPDH (Abcam; #ab9484), goat anti-actin (Santa Cruz, #1615), mouse anti-GST (GenScript; #A00865), and mouse anti-V5 (Life Technologies; #R960-25).

    Techniques: Isolation, Sequencing, Software

    ( A ) Naa10 +/Y , Naa10 -/Y , and Naa10 tm1a/Y embryos at E10.5. Growth retardation (5/33, more than five somites lower or undersized compared to littermate controls), kinky trunk, and developmental arrest are shown in Naa10 -/Y (4/33) and Naa10 tm1a/Y (1/5). Scale bars: 500 μm. ( B ) The percentage lethality in newborns, comparing Naa10 wildtype (WT) ( Naa10 +/Y and Naa10 +/+ ), Naa10 -/+ and Naa10 -/Y pups until P3, derived from matings between heterozygous females and WT males. Approximately 11.6% (10/86) of WT, 24% (13/54) of Naa10 +/- , and 76.3% (29/38) of Naa10 -/Y mice were found dead before P3. ( C ) Representative images of Naa10 -/Y pups during early postnatal days compared with Naa10 +/Y . Severe developmental defects such as malformations of head and lower body (one leg; black arrowheads), whole-body edema, and anophthalmia (black arrows) are shown (N = 1 each). ( D ) Hematoxylin and eosin (H&E)-stained heart transverse section at E14.5 and vertical section at E18.5, comparing Naa10 +/Y and Naa10 -/Y embryos. Naa10 -/Y embryo shows a ventricular septal defect (VSD) at E14.5 and E18.5. Also, at E18.5, Naa10 -/Y embryo shows atrial septal defect (ASD). Arrow indicates VSD, ASD, and double outlet right ventricle (DORV). Scale bars: 20 μm.

    Journal: eLife

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    doi: 10.7554/eLife.65952

    Figure Lengend Snippet: ( A ) Naa10 +/Y , Naa10 -/Y , and Naa10 tm1a/Y embryos at E10.5. Growth retardation (5/33, more than five somites lower or undersized compared to littermate controls), kinky trunk, and developmental arrest are shown in Naa10 -/Y (4/33) and Naa10 tm1a/Y (1/5). Scale bars: 500 μm. ( B ) The percentage lethality in newborns, comparing Naa10 wildtype (WT) ( Naa10 +/Y and Naa10 +/+ ), Naa10 -/+ and Naa10 -/Y pups until P3, derived from matings between heterozygous females and WT males. Approximately 11.6% (10/86) of WT, 24% (13/54) of Naa10 +/- , and 76.3% (29/38) of Naa10 -/Y mice were found dead before P3. ( C ) Representative images of Naa10 -/Y pups during early postnatal days compared with Naa10 +/Y . Severe developmental defects such as malformations of head and lower body (one leg; black arrowheads), whole-body edema, and anophthalmia (black arrows) are shown (N = 1 each). ( D ) Hematoxylin and eosin (H&E)-stained heart transverse section at E14.5 and vertical section at E18.5, comparing Naa10 +/Y and Naa10 -/Y embryos. Naa10 -/Y embryo shows a ventricular septal defect (VSD) at E14.5 and E18.5. Also, at E18.5, Naa10 -/Y embryo shows atrial septal defect (ASD). Arrow indicates VSD, ASD, and double outlet right ventricle (DORV). Scale bars: 20 μm.

    Article Snippet: Antibody , Anti-Naa10 (rabbit monoclonal) , Cell Signaling , Cat# 13357 , (1:1000).

    Techniques: Derivative Assay, Staining

    ( A ) Wildtype male heart outflow tract region indicating separate aorta and pulmonary trunks nestled between left and right atria. ( B ) Naa10 -/- female heart from dying P0 pup only has a single outflow tract emerging from the right ventricle, resulting in persistent truncus arteriosus. ( C ) Naa10 -/y male heart from dying P0 pup has separate outflow tracts but both emerge from the right ventricle, resulting in double outlet right ventricle. ( D ) Normal histology from wildtype male heart of pulmonary artery exiting the right ventricle. ( E ) Histology from Naa10 -/- heart of single outflow tract exiting the right ventricle with tricuspid valve leaflets. ( F ) Histology from Naa10 -/y heart of both pulmonary and aortic arteries emerging from right ventricle within the same plane. ( G ) Naa10 -/- histology showing membranous ventricular septal defect (VSD). ( H ) Naa10 -/y histology showing muscular VSD. ( I ) Normal heart revealing closed ductus arterious. ( J ) Naa10 -/- histology showing open ductus arterious leading to pulmonary overload and likely lethality.

    Journal: eLife

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    doi: 10.7554/eLife.65952

    Figure Lengend Snippet: ( A ) Wildtype male heart outflow tract region indicating separate aorta and pulmonary trunks nestled between left and right atria. ( B ) Naa10 -/- female heart from dying P0 pup only has a single outflow tract emerging from the right ventricle, resulting in persistent truncus arteriosus. ( C ) Naa10 -/y male heart from dying P0 pup has separate outflow tracts but both emerge from the right ventricle, resulting in double outlet right ventricle. ( D ) Normal histology from wildtype male heart of pulmonary artery exiting the right ventricle. ( E ) Histology from Naa10 -/- heart of single outflow tract exiting the right ventricle with tricuspid valve leaflets. ( F ) Histology from Naa10 -/y heart of both pulmonary and aortic arteries emerging from right ventricle within the same plane. ( G ) Naa10 -/- histology showing membranous ventricular septal defect (VSD). ( H ) Naa10 -/y histology showing muscular VSD. ( I ) Normal heart revealing closed ductus arterious. ( J ) Naa10 -/- histology showing open ductus arterious leading to pulmonary overload and likely lethality.

    Article Snippet: Antibody , Anti-Naa10 (rabbit monoclonal) , Cell Signaling , Cat# 13357 , (1:1000).

    Techniques:

    ( A–C ) Representative images of abnormalities in Naa10 -/Y compared with Naa10 +/Y . ( A ) Body weight of male (left) and female (right) versus ages was monitored from 2 weeks. The weight of Naa10 -/Y and Naa10 -/- mice is markedly reduced compared with that of the wildtype (WT) mice. Asterisks indicate a statistical difference calculated by Student’s t -test: *p<0.05. ( B ) Representative images of completely penetrant phenotypes. Hypopigmentation ( Naa10 +/Y , n = 243; Naa10 -/Y , n = 121; Naa10 tm1a/Y , n = 17) and supernumerary ribs ( Naa10 +/Y , n = 3; Naa10 -/Y , n = 6; Naa10 tm1a/Y , n = 2; E18.5) were found 100% in Naa10 -deficient mice. ( C ) Naa10 -/Y is smaller in size and has round-shaped head ( Naa10 +/Y 0/59, Naa10 -/Y 7/33). Over time, hydrocephaly became apparent (N = 14/29 [~48%] for >P7 male Naa10 -Y ; N = 7/19 [~36%] for >P7 female Naa10 -/- ). Hydronephrosis (red arrow, Naa10 +/Y 0/23, Naa10 -/Y 14/29, Naa10 +/+ 0/5, Naa10 -/- 7/19) and abnormal genitalia (black arrow) of male (middle, Naa10 +/Y 0/23, Naa10 -/Y 16/29) and female (bottom, hydrometrocolpos, Naa10 +/+ 0/5, Naa10 -/- 7/19) are shown.

    Journal: eLife

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    doi: 10.7554/eLife.65952

    Figure Lengend Snippet: ( A–C ) Representative images of abnormalities in Naa10 -/Y compared with Naa10 +/Y . ( A ) Body weight of male (left) and female (right) versus ages was monitored from 2 weeks. The weight of Naa10 -/Y and Naa10 -/- mice is markedly reduced compared with that of the wildtype (WT) mice. Asterisks indicate a statistical difference calculated by Student’s t -test: *p<0.05. ( B ) Representative images of completely penetrant phenotypes. Hypopigmentation ( Naa10 +/Y , n = 243; Naa10 -/Y , n = 121; Naa10 tm1a/Y , n = 17) and supernumerary ribs ( Naa10 +/Y , n = 3; Naa10 -/Y , n = 6; Naa10 tm1a/Y , n = 2; E18.5) were found 100% in Naa10 -deficient mice. ( C ) Naa10 -/Y is smaller in size and has round-shaped head ( Naa10 +/Y 0/59, Naa10 -/Y 7/33). Over time, hydrocephaly became apparent (N = 14/29 [~48%] for >P7 male Naa10 -Y ; N = 7/19 [~36%] for >P7 female Naa10 -/- ). Hydronephrosis (red arrow, Naa10 +/Y 0/23, Naa10 -/Y 14/29, Naa10 +/+ 0/5, Naa10 -/- 7/19) and abnormal genitalia (black arrow) of male (middle, Naa10 +/Y 0/23, Naa10 -/Y 16/29) and female (bottom, hydrometrocolpos, Naa10 +/+ 0/5, Naa10 -/- 7/19) are shown.

    Article Snippet: Antibody , Anti-Naa10 (rabbit monoclonal) , Cell Signaling , Cat# 13357 , (1:1000).

    Techniques:

    Skeletal analyses for ribs, sternebrae, and vertebrae.

    Journal: eLife

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    doi: 10.7554/eLife.65952

    Figure Lengend Snippet: Skeletal analyses for ribs, sternebrae, and vertebrae.

    Article Snippet: Antibody , Anti-Naa10 (rabbit monoclonal) , Cell Signaling , Cat# 13357 , (1:1000).

    Techniques:

    ( A ) In wildtype (WT) mice, 13 thoracic vertebrae and ribs are numbered, whereas 14 thoracic vertebrae and ribs are counted in mutants ( Naa10 -/Y ) (WT on the left, mutant on the right). n = 11 CT scans for Naa10 -/Y compared to n = 18 Naa10 +/Y . ( B–D ) Different number of ribs are linking the sternum between in Naa10 -/Y , Naa10 -/- , and WT. ( B ) Seven ribs linking the sternum in WT. ( C ) Eight ribs linking the sternum (the white arrow shows the eighth rib) in Naa10 -/Y . ( D ) Seven on one side plus seven and one almost linking on the other side. In two mice, an asymmetrical link was observed. White arrow shows the eighth asymmetrical rib. ( E, F ) Abnormalities in the cervical phenotype. ( E ) Cervical WT/morphology. ( F ) Partial fusion of C1 and C2 dorsal arch in one mutant mouse.

    Journal: eLife

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    doi: 10.7554/eLife.65952

    Figure Lengend Snippet: ( A ) In wildtype (WT) mice, 13 thoracic vertebrae and ribs are numbered, whereas 14 thoracic vertebrae and ribs are counted in mutants ( Naa10 -/Y ) (WT on the left, mutant on the right). n = 11 CT scans for Naa10 -/Y compared to n = 18 Naa10 +/Y . ( B–D ) Different number of ribs are linking the sternum between in Naa10 -/Y , Naa10 -/- , and WT. ( B ) Seven ribs linking the sternum in WT. ( C ) Eight ribs linking the sternum (the white arrow shows the eighth rib) in Naa10 -/Y . ( D ) Seven on one side plus seven and one almost linking on the other side. In two mice, an asymmetrical link was observed. White arrow shows the eighth asymmetrical rib. ( E, F ) Abnormalities in the cervical phenotype. ( E ) Cervical WT/morphology. ( F ) Partial fusion of C1 and C2 dorsal arch in one mutant mouse.

    Article Snippet: Antibody , Anti-Naa10 (rabbit monoclonal) , Cell Signaling , Cat# 13357 , (1:1000).

    Techniques: Mutagenesis

    ( A ) Representative images and histology of renal defects at E18.5 (n = 6 out of 39 examined) and P3 (n = 4 out of 11 examined). ( B ) Hydrocephaly (n = 3 CT scans for Naa10 -/Y mice with hydrocephaly compared to n = 3 Naa10 -/Y mice without hydrocephaly). ( C ) Kaplan–Meier survival curve of the Naa10 +/Y and Naa10 -/Y male mice starting at 4 days of life, thus not including any mice that died in the first three days of life.

    Journal: eLife

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    doi: 10.7554/eLife.65952

    Figure Lengend Snippet: ( A ) Representative images and histology of renal defects at E18.5 (n = 6 out of 39 examined) and P3 (n = 4 out of 11 examined). ( B ) Hydrocephaly (n = 3 CT scans for Naa10 -/Y mice with hydrocephaly compared to n = 3 Naa10 -/Y mice without hydrocephaly). ( C ) Kaplan–Meier survival curve of the Naa10 +/Y and Naa10 -/Y male mice starting at 4 days of life, thus not including any mice that died in the first three days of life.

    Article Snippet: Antibody , Anti-Naa10 (rabbit monoclonal) , Cell Signaling , Cat# 13357 , (1:1000).

    Techniques:

    ( A ) Correlation of Naa10 alteration state on amino-terminal acetylation in mouse embryonic fibroblasts (MEFs). Each dot (n = 533) represents the average amino-terminal acetylation percentage of five replicates of Naa10 +/Y and Naa10 -/Y , respectively. Dashed lines are the borders of ±10% difference. Except for the 10 dots, 522 of the 533 dots are within the borders. The marginal histograms show the distribution of amino-terminal acetylation data points. ( B ) Immunoprecipitation of Naa15. Liver tissue from WT and Naa10 KO mouse was lysed and incubated with anti-Naa15 antibody to retrieve NatA complexes. Proteins were separated by SDS-PAGE and immunoblots probed with anti-Naa15 antibody and anti-NAA10 antibody. ( C ) Catalytic activity of immunoprecipitated NatA. The catalytic activity of NatA precipitated from WT and Naa10 KO mouse liver tissue by anti-Naa15 was measured towards the NatA substrate peptide SESS 24 in an in vitro [ 14 C]-Ac-CoA–based acetylation assay. Control reactions were performed with no enzyme or no peptide to account for background signal. The immunoprecipitation (IP) and activity measurements were performed in three independent setups, each with three technical replicates per assay. One representative setup is shown. Figure 3—source data 1. Identification of a potential Naa10 homolog.

    Journal: eLife

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    doi: 10.7554/eLife.65952

    Figure Lengend Snippet: ( A ) Correlation of Naa10 alteration state on amino-terminal acetylation in mouse embryonic fibroblasts (MEFs). Each dot (n = 533) represents the average amino-terminal acetylation percentage of five replicates of Naa10 +/Y and Naa10 -/Y , respectively. Dashed lines are the borders of ±10% difference. Except for the 10 dots, 522 of the 533 dots are within the borders. The marginal histograms show the distribution of amino-terminal acetylation data points. ( B ) Immunoprecipitation of Naa15. Liver tissue from WT and Naa10 KO mouse was lysed and incubated with anti-Naa15 antibody to retrieve NatA complexes. Proteins were separated by SDS-PAGE and immunoblots probed with anti-Naa15 antibody and anti-NAA10 antibody. ( C ) Catalytic activity of immunoprecipitated NatA. The catalytic activity of NatA precipitated from WT and Naa10 KO mouse liver tissue by anti-Naa15 was measured towards the NatA substrate peptide SESS 24 in an in vitro [ 14 C]-Ac-CoA–based acetylation assay. Control reactions were performed with no enzyme or no peptide to account for background signal. The immunoprecipitation (IP) and activity measurements were performed in three independent setups, each with three technical replicates per assay. One representative setup is shown. Figure 3—source data 1. Identification of a potential Naa10 homolog.

    Article Snippet: Antibody , Anti-Naa10 (rabbit monoclonal) , Cell Signaling , Cat# 13357 , (1:1000).

    Techniques: Immunoprecipitation, Incubation, SDS Page, Western Blot, Activity Assay, In Vitro, Acetylation Assay

    ( A ) Expression of Naa10 and Naa12 in wildtype (WT), Naa10 KO and Naa12 KO tissues, adult mice 13 weeks of age, (brain, heart, kidney, and testis) by RT-PCR. Expression of GAPDH was analyzed as an endogenous control. ( B ) Phenotypes in Naa12 KO mice. Lack of hypopigmentation (upper; N = 29) and lack of supernumerary ribs (middle and bottom; E18.5; N = 7) in Naa12 KO mouse.

    Journal: eLife

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    doi: 10.7554/eLife.65952

    Figure Lengend Snippet: ( A ) Expression of Naa10 and Naa12 in wildtype (WT), Naa10 KO and Naa12 KO tissues, adult mice 13 weeks of age, (brain, heart, kidney, and testis) by RT-PCR. Expression of GAPDH was analyzed as an endogenous control. ( B ) Phenotypes in Naa12 KO mice. Lack of hypopigmentation (upper; N = 29) and lack of supernumerary ribs (middle and bottom; E18.5; N = 7) in Naa12 KO mouse.

    Article Snippet: Antibody , Anti-Naa10 (rabbit monoclonal) , Cell Signaling , Cat# 13357 , (1:1000).

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction

    ( A ) In vitro N-terminal acetyltransferase radioactive-based assay. Comparison of mouse Naa10 and Naa12 towards Naa10 peptide substrates, beta-actin (DDDIA-) and gamma-actin (EEEIA-), and the optimal NatA complex peptide substrate, SESSS-. Background control reactions were performed in the absence of either peptide or enzyme. Assays were performed in triplicate; error bars represent SEM. ( B ) Co-immunoprecipitation assay. HEK293 cells were transfected as indicated and lysed after 48 hr. Cell lysates were incubated with 1 μg anti-V5 antibody to precipitate V5-tagged Naa15. The isolated complexes were separated on SDS-PAGE and probed with the indicated antibodies. ( C ) Recombinant mouse Naa12/human Naa15 chimera complex activity. Radioactive acetyltransferase activity assay evaluating the activity of mNaa12-hNaa15 towards peptide (closed circles, ‘mNaa12-hNaa15’) and peptide chemical acetylation in the absence of enzyme (closed circles, ‘Buffer’) as well as chemical acetylation of the enzyme in the absence of peptide (open circles) assay and background (open circles). Error bars represent SD of two technical replicates. These are the same results from fraction #14 (both SESSS- and No Peptide) and both Buffer and Background used to illustrate the size-exclusion-purified mNaa12-hNaa15 complex activity in . Figure 4—source data 1. Characterization of a potential Naa10 homolog.

    Journal: eLife

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    doi: 10.7554/eLife.65952

    Figure Lengend Snippet: ( A ) In vitro N-terminal acetyltransferase radioactive-based assay. Comparison of mouse Naa10 and Naa12 towards Naa10 peptide substrates, beta-actin (DDDIA-) and gamma-actin (EEEIA-), and the optimal NatA complex peptide substrate, SESSS-. Background control reactions were performed in the absence of either peptide or enzyme. Assays were performed in triplicate; error bars represent SEM. ( B ) Co-immunoprecipitation assay. HEK293 cells were transfected as indicated and lysed after 48 hr. Cell lysates were incubated with 1 μg anti-V5 antibody to precipitate V5-tagged Naa15. The isolated complexes were separated on SDS-PAGE and probed with the indicated antibodies. ( C ) Recombinant mouse Naa12/human Naa15 chimera complex activity. Radioactive acetyltransferase activity assay evaluating the activity of mNaa12-hNaa15 towards peptide (closed circles, ‘mNaa12-hNaa15’) and peptide chemical acetylation in the absence of enzyme (closed circles, ‘Buffer’) as well as chemical acetylation of the enzyme in the absence of peptide (open circles) assay and background (open circles). Error bars represent SD of two technical replicates. These are the same results from fraction #14 (both SESSS- and No Peptide) and both Buffer and Background used to illustrate the size-exclusion-purified mNaa12-hNaa15 complex activity in . Figure 4—source data 1. Characterization of a potential Naa10 homolog.

    Article Snippet: Antibody , Anti-Naa10 (rabbit monoclonal) , Cell Signaling , Cat# 13357 , (1:1000).

    Techniques: In Vitro, Radioactivity, Co-Immunoprecipitation Assay, Transfection, Incubation, Isolation, SDS Page, Recombinant, Activity Assay, Purification

     Naa10,  Naa11, and Naa12 peptides identified by LC-MS/MS analysis in Naa15 IP samples from WT and Naa10 -KO mouse.

    Journal: eLife

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    doi: 10.7554/eLife.65952

    Figure Lengend Snippet: Naa10, Naa11, and Naa12 peptides identified by LC-MS/MS analysis in Naa15 IP samples from WT and Naa10 -KO mouse.

    Article Snippet: Antibody , Anti-Naa10 (rabbit monoclonal) , Cell Signaling , Cat# 13357 , (1:1000).

    Techniques: Sequencing

    Naa10 Naa12 DKO exhibit embryonic lethality. Pedigree and genotypes of pups and embryos at E10.5 and E18.5 from Naa10 +/- Naa12 +/- female mice crossed to the Naa10 +/Y Naa12 +/- male mice.

    Journal: eLife

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    doi: 10.7554/eLife.65952

    Figure Lengend Snippet: Naa10 Naa12 DKO exhibit embryonic lethality. Pedigree and genotypes of pups and embryos at E10.5 and E18.5 from Naa10 +/- Naa12 +/- female mice crossed to the Naa10 +/Y Naa12 +/- male mice.

    Article Snippet: Antibody , Anti-Naa10 (rabbit monoclonal) , Cell Signaling , Cat# 13357 , (1:1000).

    Techniques:

    Litter size of  Naa10  × Naa12 matings.

    Journal: eLife

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    doi: 10.7554/eLife.65952

    Figure Lengend Snippet: Litter size of Naa10 × Naa12 matings.

    Article Snippet: Antibody , Anti-Naa10 (rabbit monoclonal) , Cell Signaling , Cat# 13357 , (1:1000).

    Techniques:

    Naa10 Naa12 double-knockout (DKO) exhibit embryonic lethality. Pedigree of mating and genotypes of pups and embryos at E8.5, E10.5, E12.5, and E18.5.

    Journal: eLife

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    doi: 10.7554/eLife.65952

    Figure Lengend Snippet: Naa10 Naa12 double-knockout (DKO) exhibit embryonic lethality. Pedigree of mating and genotypes of pups and embryos at E8.5, E10.5, E12.5, and E18.5.

    Article Snippet: Antibody , Anti-Naa10 (rabbit monoclonal) , Cell Signaling , Cat# 13357 , (1:1000).

    Techniques: Double Knockout

    ( A ) Male body weight for the Naa10 mice on inbred genetic background (eight backcrosses to C57bl6/J). ( B ) Female body weight for the Naa10 mice on inbred genetic background (eight backcrosses to C57bl6/J). ( C ) Male body weight for the Naa10 and Naa12 mice on mixed genetic background. ( D ) Female body weight for the Naa10 and Naa12 mice on mixed genetic background.

    Journal: eLife

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    doi: 10.7554/eLife.65952

    Figure Lengend Snippet: ( A ) Male body weight for the Naa10 mice on inbred genetic background (eight backcrosses to C57bl6/J). ( B ) Female body weight for the Naa10 mice on inbred genetic background (eight backcrosses to C57bl6/J). ( C ) Male body weight for the Naa10 and Naa12 mice on mixed genetic background. ( D ) Female body weight for the Naa10 and Naa12 mice on mixed genetic background.

    Article Snippet: Antibody , Anti-Naa10 (rabbit monoclonal) , Cell Signaling , Cat# 13357 , (1:1000).

    Techniques:

    Model D 4 genotype survival by age.

    Journal: eLife

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    doi: 10.7554/eLife.65952

    Figure Lengend Snippet: Model D 4 genotype survival by age.

    Article Snippet: Antibody , Anti-Naa10 (rabbit monoclonal) , Cell Signaling , Cat# 13357 , (1:1000).

    Techniques:

    Journal: eLife

    Article Title: Naa12 compensates for Naa10 in mice in the amino-terminal acetylation pathway

    doi: 10.7554/eLife.65952

    Figure Lengend Snippet:

    Article Snippet: Antibody , Anti-Naa10 (rabbit monoclonal) , Cell Signaling , Cat# 13357 , (1:1000).

    Techniques: Isolation, Sequencing, Software

    a hNAA50(magenta), hNAA15(green), and hNAA10(orange) within the hNatE complex overlay with ScNatE (gray, PDB: 6O07). The α3 helix and β7 strand of Naa50 is shown to shift toward Naa10 in the human structure ( b ) hNatE aligned with hNatA (light blue, PDB:6C9M) and hNAA50 (wheat, PDB: 3TFY). The top zoom-in area shows the alignment of free hNAA50 and hNatE. The below zoom-in area shows the hNAA10 conformational change induced by hNAA50 binding.

    Journal: Nature Communications

    Article Title: Molecular basis for N-terminal acetylation by human NatE and its modulation by HYPK

    doi: 10.1038/s41467-020-14584-7

    Figure Lengend Snippet: a hNAA50(magenta), hNAA15(green), and hNAA10(orange) within the hNatE complex overlay with ScNatE (gray, PDB: 6O07). The α3 helix and β7 strand of Naa50 is shown to shift toward Naa10 in the human structure ( b ) hNatE aligned with hNatA (light blue, PDB:6C9M) and hNAA50 (wheat, PDB: 3TFY). The top zoom-in area shows the alignment of free hNAA50 and hNatE. The below zoom-in area shows the hNAA10 conformational change induced by hNAA50 binding.

    Article Snippet: Proteins were separated by SDS–PAGE, transferred onto a nitrocellulose membrane (Amersham Protran 0.2 µM NC), and probed with V5-tag mouse monoclonal antibody (1:5000, Invitrogen, R960-25) and NAA10 rabbit monoclonal antibody (1:1000, Cell Signaling #13357).

    Techniques: Binding Assay