25309 Search Results


prosthecobacter  (ATCC)
90
ATCC prosthecobacter
Prosthecobacter, supplied by ATCC, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 90 stars, based on 1 article reviews
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mouse monoclonal anti fxr nr1h4 d 3  (Santa Cruz Biotechnology)
96
Santa Cruz Biotechnology mouse monoclonal anti fxr nr1h4 d 3

Mouse Monoclonal Anti Fxr Nr1h4 D 3, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/mouse monoclonal anti fxr nr1h4 d 3/product/Santa Cruz Biotechnology
Average 96 stars, based on 1 article reviews
mouse monoclonal anti fxr nr1h4 d 3 - by Bioz Stars, 2025-07
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rabbit polyclonal anti pih1d3  (Proteintech)
92
Proteintech rabbit polyclonal anti pih1d3
Deletion of the <t>Pih1d3</t> gene in knockout rats causes growth retardation and premature death. (A) Schematic showing the structure of the rat Pih1d3 gene with the region targeted for mutagenesis by TALEN and with the region amplified by PCR for genotyping and sequencing (gene structure not drawn to scale). Partial sequence of exon 5 on the rat Pih1d3 gene shows the binding sites for the left (TALEN-L) and right (TALEN-R) arms of an effective TALEN as well as the sequence deleted in the Pih1d3-KO rats. Deletion of 10 nucleotides from exon 5 results in the shifting of the open reading frame, introducing two consecutive stop codons within exon 6 of the rat Pih1d3 gene (sequence verified, but data not shown). (B) Immunoblotting revealed that the PIH1D3 protein was depleted from the tissues of Pih1d3-KO rats. Equal loading of total proteins (20 µg/lane) was assessed by immunoblotting on the same membrane for actin. Tissues were dissected from Pih1d3-KO male rats and their WT male littermates at the age of 15 days. (C) A representative image shows that a Pih1d3-KO rat had a smaller body but an enlarged head compared to its male WT littermate at the age of 28 days. (D) Growth chart showing the retarded growth of body weight (g, gram) in Pih1d3-KO rats compared to their male WT littermates (data were the means ± standard deviation). (E) Kaplan–Meier survival analysis showing the premature death of Pih1d3-KO rats during the postnatal development stage. Very few KO rats survived beyond 30 days, but none of them survived by 70 days of ages.
Rabbit Polyclonal Anti Pih1d3, supplied by Proteintech, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rabbit polyclonal anti pih1d3/product/Proteintech
Average 92 stars, based on 1 article reviews
rabbit polyclonal anti pih1d3 - by Bioz Stars, 2025-07
92/100 stars
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santa cruz biotechnology sc 25309  (Santa Cruz Biotechnology)
86
Santa Cruz Biotechnology santa cruz biotechnology sc 25309
Deletion of the <t>Pih1d3</t> gene in knockout rats causes growth retardation and premature death. (A) Schematic showing the structure of the rat Pih1d3 gene with the region targeted for mutagenesis by TALEN and with the region amplified by PCR for genotyping and sequencing (gene structure not drawn to scale). Partial sequence of exon 5 on the rat Pih1d3 gene shows the binding sites for the left (TALEN-L) and right (TALEN-R) arms of an effective TALEN as well as the sequence deleted in the Pih1d3-KO rats. Deletion of 10 nucleotides from exon 5 results in the shifting of the open reading frame, introducing two consecutive stop codons within exon 6 of the rat Pih1d3 gene (sequence verified, but data not shown). (B) Immunoblotting revealed that the PIH1D3 protein was depleted from the tissues of Pih1d3-KO rats. Equal loading of total proteins (20 µg/lane) was assessed by immunoblotting on the same membrane for actin. Tissues were dissected from Pih1d3-KO male rats and their WT male littermates at the age of 15 days. (C) A representative image shows that a Pih1d3-KO rat had a smaller body but an enlarged head compared to its male WT littermate at the age of 28 days. (D) Growth chart showing the retarded growth of body weight (g, gram) in Pih1d3-KO rats compared to their male WT littermates (data were the means ± standard deviation). (E) Kaplan–Meier survival analysis showing the premature death of Pih1d3-KO rats during the postnatal development stage. Very few KO rats survived beyond 30 days, but none of them survived by 70 days of ages.
Santa Cruz Biotechnology Sc 25309, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/santa cruz biotechnology sc 25309/product/Santa Cruz Biotechnology
Average 86 stars, based on 1 article reviews
santa cruz biotechnology sc 25309 - by Bioz Stars, 2025-07
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25309 25317 technology development co  (Technology Development Co)
86
Technology Development Co 25309 25317 technology development co
Deletion of the <t>Pih1d3</t> gene in knockout rats causes growth retardation and premature death. (A) Schematic showing the structure of the rat Pih1d3 gene with the region targeted for mutagenesis by TALEN and with the region amplified by PCR for genotyping and sequencing (gene structure not drawn to scale). Partial sequence of exon 5 on the rat Pih1d3 gene shows the binding sites for the left (TALEN-L) and right (TALEN-R) arms of an effective TALEN as well as the sequence deleted in the Pih1d3-KO rats. Deletion of 10 nucleotides from exon 5 results in the shifting of the open reading frame, introducing two consecutive stop codons within exon 6 of the rat Pih1d3 gene (sequence verified, but data not shown). (B) Immunoblotting revealed that the PIH1D3 protein was depleted from the tissues of Pih1d3-KO rats. Equal loading of total proteins (20 µg/lane) was assessed by immunoblotting on the same membrane for actin. Tissues were dissected from Pih1d3-KO male rats and their WT male littermates at the age of 15 days. (C) A representative image shows that a Pih1d3-KO rat had a smaller body but an enlarged head compared to its male WT littermate at the age of 28 days. (D) Growth chart showing the retarded growth of body weight (g, gram) in Pih1d3-KO rats compared to their male WT littermates (data were the means ± standard deviation). (E) Kaplan–Meier survival analysis showing the premature death of Pih1d3-KO rats during the postnatal development stage. Very few KO rats survived beyond 30 days, but none of them survived by 70 days of ages.
25309 25317 Technology Development Co, supplied by Technology Development Co, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/25309 25317 technology development co/product/Technology Development Co
Average 86 stars, based on 1 article reviews
25309 25317 technology development co - by Bioz Stars, 2025-07
86/100 stars
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Image Search Results


Journal: iScience

Article Title: Hepatocyte activity of the cholesterol sensor smoothened regulates cholesterol and bile acid homeostasis in mice

doi: 10.1016/j.isci.2021.103089

Figure Lengend Snippet:

Article Snippet: Mouse monoclonal anti-FXR/NR1H4 (D-3) , Santa Cruz , Cat#sc-25309; RRID: AB_628039.

Techniques: Plasmid Preparation, Recombinant, Western Blot, Protease Inhibitor, Stripping, Quantitation Assay, Colorimetric Assay, Bicinchoninic Acid Protein Assay, SYBR Green Assay, Software, Imaging, Real-time Polymerase Chain Reaction

Deletion of the Pih1d3 gene in knockout rats causes growth retardation and premature death. (A) Schematic showing the structure of the rat Pih1d3 gene with the region targeted for mutagenesis by TALEN and with the region amplified by PCR for genotyping and sequencing (gene structure not drawn to scale). Partial sequence of exon 5 on the rat Pih1d3 gene shows the binding sites for the left (TALEN-L) and right (TALEN-R) arms of an effective TALEN as well as the sequence deleted in the Pih1d3-KO rats. Deletion of 10 nucleotides from exon 5 results in the shifting of the open reading frame, introducing two consecutive stop codons within exon 6 of the rat Pih1d3 gene (sequence verified, but data not shown). (B) Immunoblotting revealed that the PIH1D3 protein was depleted from the tissues of Pih1d3-KO rats. Equal loading of total proteins (20 µg/lane) was assessed by immunoblotting on the same membrane for actin. Tissues were dissected from Pih1d3-KO male rats and their WT male littermates at the age of 15 days. (C) A representative image shows that a Pih1d3-KO rat had a smaller body but an enlarged head compared to its male WT littermate at the age of 28 days. (D) Growth chart showing the retarded growth of body weight (g, gram) in Pih1d3-KO rats compared to their male WT littermates (data were the means ± standard deviation). (E) Kaplan–Meier survival analysis showing the premature death of Pih1d3-KO rats during the postnatal development stage. Very few KO rats survived beyond 30 days, but none of them survived by 70 days of ages.

Journal: Frontiers in Cell and Developmental Biology

Article Title: PIH1D3-knockout rats exhibit full ciliopathy features and dysfunctional pre-assembly and loading of dynein arms in motile cilia

doi: 10.3389/fcell.2023.1282787

Figure Lengend Snippet: Deletion of the Pih1d3 gene in knockout rats causes growth retardation and premature death. (A) Schematic showing the structure of the rat Pih1d3 gene with the region targeted for mutagenesis by TALEN and with the region amplified by PCR for genotyping and sequencing (gene structure not drawn to scale). Partial sequence of exon 5 on the rat Pih1d3 gene shows the binding sites for the left (TALEN-L) and right (TALEN-R) arms of an effective TALEN as well as the sequence deleted in the Pih1d3-KO rats. Deletion of 10 nucleotides from exon 5 results in the shifting of the open reading frame, introducing two consecutive stop codons within exon 6 of the rat Pih1d3 gene (sequence verified, but data not shown). (B) Immunoblotting revealed that the PIH1D3 protein was depleted from the tissues of Pih1d3-KO rats. Equal loading of total proteins (20 µg/lane) was assessed by immunoblotting on the same membrane for actin. Tissues were dissected from Pih1d3-KO male rats and their WT male littermates at the age of 15 days. (C) A representative image shows that a Pih1d3-KO rat had a smaller body but an enlarged head compared to its male WT littermate at the age of 28 days. (D) Growth chart showing the retarded growth of body weight (g, gram) in Pih1d3-KO rats compared to their male WT littermates (data were the means ± standard deviation). (E) Kaplan–Meier survival analysis showing the premature death of Pih1d3-KO rats during the postnatal development stage. Very few KO rats survived beyond 30 days, but none of them survived by 70 days of ages.

Article Snippet: The following primary antibodies were used: rabbit polyclonal anti-PIH1D3 (Proteintech, 25309-1-AP); mouse monoclonal anti-acetylated alpha-tubulin IgG1 (Proteintech, CL488-66200); rabbit polyclonal anti-DNAI1 (Abcam, ab171964); rabbit polyclonal anti-DNAI2 (Proteintech, 17533-1-AP); rabbit polyclonal anti-DNALI1 for immunofluorescence staining (Proteintech, 17601-1-AP); mouse monoclonal anti-DNALI1 for immunoblotting (Santa Cruz, Sc-514831); rabbit polyclonal anti-IFT52 (Proteintech, 17534-1-AP); rabbit polyclonal anti-IFT57 (Proteintech, 11083-1-AP); rabbit polyclonal anti-PIH1D1 (Proteintech, 19427-1-AP); rabbit polyclonal anti-IFT20 (Proteintech, 13615-1-AP); rabbit polyclonal anti-IFT81 (Proteintech, 11744-1-AP); mouse monoclonal anti-Flag (Novus, NBP1-97410); and mouse monoclonal anti-Myc (Invitrogen, MA1-21316).

Techniques: Knock-Out, Mutagenesis, Amplification, Sequencing, Binding Assay, Western Blot, Membrane, Standard Deviation

Communicating hydrocephalus detected in Pih1d3-KO rats. (A–H) Immunofluorescent staining for acetylated alpha-tubulin (acetyl-TUBA1A, a marker for motile cilia) and PIH1D3 reveals the depletion of PIH1D3 proteins from ependymal cells in Pih1d3-KO rats. KO rats and WT male littermates were examined at the age of 5 days. Scale bars: 100 µm (A,E) and 30 µm (B–D,F–H) . (I,J) Images of Pih1d3-KO and WT rat brains at postnatal day 5. Arrows point to the enlarged and transparent brain hemispheres. (K–P) HE staining of coronal brain sections reveals enlarged lateral ventricles (LV). Scale bar: 150 µm. (Q,R) HE staining of sagittal brain sections reveals no blocking of the aqueduct (aq). (I-J) The same rat brains were sectioned for HE staining (K–R) . (S,T) CSF flow analysis reveals that Evans blue injected into the left LV flowed into the right LV but not into the dorsal third ventricle (D3V), aqueduct, and the fourth ventricle (fourth V) in Pih1d3-KO rats. By contrast, Evans blue flowed from the left LV into D3V, aqueduct, and then fourth V but not into the right LV in wild-type rats. Scale bar: 200 µm.

Journal: Frontiers in Cell and Developmental Biology

Article Title: PIH1D3-knockout rats exhibit full ciliopathy features and dysfunctional pre-assembly and loading of dynein arms in motile cilia

doi: 10.3389/fcell.2023.1282787

Figure Lengend Snippet: Communicating hydrocephalus detected in Pih1d3-KO rats. (A–H) Immunofluorescent staining for acetylated alpha-tubulin (acetyl-TUBA1A, a marker for motile cilia) and PIH1D3 reveals the depletion of PIH1D3 proteins from ependymal cells in Pih1d3-KO rats. KO rats and WT male littermates were examined at the age of 5 days. Scale bars: 100 µm (A,E) and 30 µm (B–D,F–H) . (I,J) Images of Pih1d3-KO and WT rat brains at postnatal day 5. Arrows point to the enlarged and transparent brain hemispheres. (K–P) HE staining of coronal brain sections reveals enlarged lateral ventricles (LV). Scale bar: 150 µm. (Q,R) HE staining of sagittal brain sections reveals no blocking of the aqueduct (aq). (I-J) The same rat brains were sectioned for HE staining (K–R) . (S,T) CSF flow analysis reveals that Evans blue injected into the left LV flowed into the right LV but not into the dorsal third ventricle (D3V), aqueduct, and the fourth ventricle (fourth V) in Pih1d3-KO rats. By contrast, Evans blue flowed from the left LV into D3V, aqueduct, and then fourth V but not into the right LV in wild-type rats. Scale bar: 200 µm.

Article Snippet: The following primary antibodies were used: rabbit polyclonal anti-PIH1D3 (Proteintech, 25309-1-AP); mouse monoclonal anti-acetylated alpha-tubulin IgG1 (Proteintech, CL488-66200); rabbit polyclonal anti-DNAI1 (Abcam, ab171964); rabbit polyclonal anti-DNAI2 (Proteintech, 17533-1-AP); rabbit polyclonal anti-DNALI1 for immunofluorescence staining (Proteintech, 17601-1-AP); mouse monoclonal anti-DNALI1 for immunoblotting (Santa Cruz, Sc-514831); rabbit polyclonal anti-IFT52 (Proteintech, 17534-1-AP); rabbit polyclonal anti-IFT57 (Proteintech, 11083-1-AP); rabbit polyclonal anti-PIH1D1 (Proteintech, 19427-1-AP); rabbit polyclonal anti-IFT20 (Proteintech, 13615-1-AP); rabbit polyclonal anti-IFT81 (Proteintech, 11744-1-AP); mouse monoclonal anti-Flag (Novus, NBP1-97410); and mouse monoclonal anti-Myc (Invitrogen, MA1-21316).

Techniques: Staining, Marker, Blocking Assay, Injection

Accumulation of mucus in the upper respiratory tract revealed in Pih1d3-knockout rats. (A–C) Immunofluorescent staining for acetylated alpha-tubulin (acetyl-TUBA1A) and PIH1D3 reveals the depletion of PIH1D3 proteins from the epithelial cells of respiratory tracts in Pih1d3-KO rats compared to WT littermates. Scale bar: 40 µm (A–C) . (D–I) HE staining reveals that mucus accumulated severely in the nasal cavity (arrows pointed to) and barely in the trachea and bronchus of Pih1d3-KO rats, but not in the WT rats. KO rats and male littermates were examined at the age of 5 days. Scale bar: 200 µm.

Journal: Frontiers in Cell and Developmental Biology

Article Title: PIH1D3-knockout rats exhibit full ciliopathy features and dysfunctional pre-assembly and loading of dynein arms in motile cilia

doi: 10.3389/fcell.2023.1282787

Figure Lengend Snippet: Accumulation of mucus in the upper respiratory tract revealed in Pih1d3-knockout rats. (A–C) Immunofluorescent staining for acetylated alpha-tubulin (acetyl-TUBA1A) and PIH1D3 reveals the depletion of PIH1D3 proteins from the epithelial cells of respiratory tracts in Pih1d3-KO rats compared to WT littermates. Scale bar: 40 µm (A–C) . (D–I) HE staining reveals that mucus accumulated severely in the nasal cavity (arrows pointed to) and barely in the trachea and bronchus of Pih1d3-KO rats, but not in the WT rats. KO rats and male littermates were examined at the age of 5 days. Scale bar: 200 µm.

Article Snippet: The following primary antibodies were used: rabbit polyclonal anti-PIH1D3 (Proteintech, 25309-1-AP); mouse monoclonal anti-acetylated alpha-tubulin IgG1 (Proteintech, CL488-66200); rabbit polyclonal anti-DNAI1 (Abcam, ab171964); rabbit polyclonal anti-DNAI2 (Proteintech, 17533-1-AP); rabbit polyclonal anti-DNALI1 for immunofluorescence staining (Proteintech, 17601-1-AP); mouse monoclonal anti-DNALI1 for immunoblotting (Santa Cruz, Sc-514831); rabbit polyclonal anti-IFT52 (Proteintech, 17534-1-AP); rabbit polyclonal anti-IFT57 (Proteintech, 11083-1-AP); rabbit polyclonal anti-PIH1D1 (Proteintech, 19427-1-AP); rabbit polyclonal anti-IFT20 (Proteintech, 13615-1-AP); rabbit polyclonal anti-IFT81 (Proteintech, 11744-1-AP); mouse monoclonal anti-Flag (Novus, NBP1-97410); and mouse monoclonal anti-Myc (Invitrogen, MA1-21316).

Techniques: Knock-Out, Staining

Spermatogenesis impaired in Pih1d3-KO rats. (A,B) Images of the testis taken from Pih1d3-KO and WT rats at the age of 28 days. Scale bar: 500 µm. (C) Immunoblotting reveals the expression of PIH1D3 in the rat testis from postnatal day 7 onward. (D) Immunofluorescent staining reveals PIH1D3 depleted from the spermatogenic cells of the KO rat testis. Scale bar: 40 µm. (E) HE staining of seminiferous tubules from KO and WT littermates at varying postnatal (P) ages. At postnatal day 14 (P14), germ cells in both the WT and KO rats were developed into pachytene spermatocytes, and no difference was observed between KO and WT rats. By P27, seminiferous tubules were filled with round spermatids (red arrow) and pachytene spermatocytes (dark arrow) in WT rats, but much fewer round spermatids were observed in KO rats. By P33, elongated spermatids (red arrowhead) were observed in the tubules of WT rats, but not of KO rats. The total number of cells, including pachytene spermatocytes and round spermatids, was significantly decreased in KO rats compared to WT littermates by P33. Scale bar: 50 µm. (F–H) TUNEL staining revealed that apoptotic cell death was increased in the seminiferous tubules of KO rats aged P28 compared to WT littermates. Data were the means ± SD (n = 50). ** p < 0.01. (I) Immunohistochemistry reveals that PIH1D3 was depleted from the epithelium of efferent ductules in KO rats. (J) HE staining reveals the occlusion and agglutination of efferent ductules and the abnormal ducts with a narrow lumen in KO rats compared to WT rats at age P35. Few spermatozoa with long tails (arrow) were observed in WT but not in KO rats. Scale bar: 40 µm (F,G,I,J) .

Journal: Frontiers in Cell and Developmental Biology

Article Title: PIH1D3-knockout rats exhibit full ciliopathy features and dysfunctional pre-assembly and loading of dynein arms in motile cilia

doi: 10.3389/fcell.2023.1282787

Figure Lengend Snippet: Spermatogenesis impaired in Pih1d3-KO rats. (A,B) Images of the testis taken from Pih1d3-KO and WT rats at the age of 28 days. Scale bar: 500 µm. (C) Immunoblotting reveals the expression of PIH1D3 in the rat testis from postnatal day 7 onward. (D) Immunofluorescent staining reveals PIH1D3 depleted from the spermatogenic cells of the KO rat testis. Scale bar: 40 µm. (E) HE staining of seminiferous tubules from KO and WT littermates at varying postnatal (P) ages. At postnatal day 14 (P14), germ cells in both the WT and KO rats were developed into pachytene spermatocytes, and no difference was observed between KO and WT rats. By P27, seminiferous tubules were filled with round spermatids (red arrow) and pachytene spermatocytes (dark arrow) in WT rats, but much fewer round spermatids were observed in KO rats. By P33, elongated spermatids (red arrowhead) were observed in the tubules of WT rats, but not of KO rats. The total number of cells, including pachytene spermatocytes and round spermatids, was significantly decreased in KO rats compared to WT littermates by P33. Scale bar: 50 µm. (F–H) TUNEL staining revealed that apoptotic cell death was increased in the seminiferous tubules of KO rats aged P28 compared to WT littermates. Data were the means ± SD (n = 50). ** p < 0.01. (I) Immunohistochemistry reveals that PIH1D3 was depleted from the epithelium of efferent ductules in KO rats. (J) HE staining reveals the occlusion and agglutination of efferent ductules and the abnormal ducts with a narrow lumen in KO rats compared to WT rats at age P35. Few spermatozoa with long tails (arrow) were observed in WT but not in KO rats. Scale bar: 40 µm (F,G,I,J) .

Article Snippet: The following primary antibodies were used: rabbit polyclonal anti-PIH1D3 (Proteintech, 25309-1-AP); mouse monoclonal anti-acetylated alpha-tubulin IgG1 (Proteintech, CL488-66200); rabbit polyclonal anti-DNAI1 (Abcam, ab171964); rabbit polyclonal anti-DNAI2 (Proteintech, 17533-1-AP); rabbit polyclonal anti-DNALI1 for immunofluorescence staining (Proteintech, 17601-1-AP); mouse monoclonal anti-DNALI1 for immunoblotting (Santa Cruz, Sc-514831); rabbit polyclonal anti-IFT52 (Proteintech, 17534-1-AP); rabbit polyclonal anti-IFT57 (Proteintech, 11083-1-AP); rabbit polyclonal anti-PIH1D1 (Proteintech, 19427-1-AP); rabbit polyclonal anti-IFT20 (Proteintech, 13615-1-AP); rabbit polyclonal anti-IFT81 (Proteintech, 11744-1-AP); mouse monoclonal anti-Flag (Novus, NBP1-97410); and mouse monoclonal anti-Myc (Invitrogen, MA1-21316).

Techniques: Western Blot, Expressing, Staining, TUNEL Assay, Immunohistochemistry, Agglutination

PIH1D3 deficiency results in the depletion of DNAI1, DNAI2, and DNALI1 from motile cilia. (A–F) Immunofluorescent staining assessed the colocalization of acetylated alpha-tubulin (acetyl-TUBA1A, a marker for motile cilia) with DNALI1 (marker for inner dynein arms) and with DNAI1 and DNAI2 (markers for outer dynein arms) in the brain ependymal cells and nasal epithelial cells of Pih1d3-KO rats and WT littermates. Acetyl-TUBA1A was well colocalized with DNALI1, DNAI1, and DNAI2 in WT rats but not in the KO rats. Brain tissues were taken from rat embryos at E19, and nasal tissues were taken from postnatal rats at P5. Scale bar: 30 µm.

Journal: Frontiers in Cell and Developmental Biology

Article Title: PIH1D3-knockout rats exhibit full ciliopathy features and dysfunctional pre-assembly and loading of dynein arms in motile cilia

doi: 10.3389/fcell.2023.1282787

Figure Lengend Snippet: PIH1D3 deficiency results in the depletion of DNAI1, DNAI2, and DNALI1 from motile cilia. (A–F) Immunofluorescent staining assessed the colocalization of acetylated alpha-tubulin (acetyl-TUBA1A, a marker for motile cilia) with DNALI1 (marker for inner dynein arms) and with DNAI1 and DNAI2 (markers for outer dynein arms) in the brain ependymal cells and nasal epithelial cells of Pih1d3-KO rats and WT littermates. Acetyl-TUBA1A was well colocalized with DNALI1, DNAI1, and DNAI2 in WT rats but not in the KO rats. Brain tissues were taken from rat embryos at E19, and nasal tissues were taken from postnatal rats at P5. Scale bar: 30 µm.

Article Snippet: The following primary antibodies were used: rabbit polyclonal anti-PIH1D3 (Proteintech, 25309-1-AP); mouse monoclonal anti-acetylated alpha-tubulin IgG1 (Proteintech, CL488-66200); rabbit polyclonal anti-DNAI1 (Abcam, ab171964); rabbit polyclonal anti-DNAI2 (Proteintech, 17533-1-AP); rabbit polyclonal anti-DNALI1 for immunofluorescence staining (Proteintech, 17601-1-AP); mouse monoclonal anti-DNALI1 for immunoblotting (Santa Cruz, Sc-514831); rabbit polyclonal anti-IFT52 (Proteintech, 17534-1-AP); rabbit polyclonal anti-IFT57 (Proteintech, 11083-1-AP); rabbit polyclonal anti-PIH1D1 (Proteintech, 19427-1-AP); rabbit polyclonal anti-IFT20 (Proteintech, 13615-1-AP); rabbit polyclonal anti-IFT81 (Proteintech, 11744-1-AP); mouse monoclonal anti-Flag (Novus, NBP1-97410); and mouse monoclonal anti-Myc (Invitrogen, MA1-21316).

Techniques: Staining, Marker

Defective ODA and IDA in the motile cilia of PIH1D3-KO rats. (A–C) Immunofluorescent staining reveals that PIH1D3 deficiency in KO rats results in the dislocation of DNAI1 and DNALI1 from motile cilia labeled with acetyl-TUBA1A. The efferent ductules were dissected from Pih1d3-KO rats and WT littermates at the age of 27 days. Scale bar: 30 µm. (D) Immunoblotting shows that the expression of the ODA markers DNAI1 and DNAI2 and the IDA marker DNALI1 was reduced in the tissues of Pih1d3-KO rats compared to WT littermates. Rats of age postnatal day 20 were used for dissecting the trachea and testis, and rats of age postnatal day 0 were used for dissecting brains. Equal loading of 20 µg total protein per lane was verified by immunoblotting for actin. Quantification for the density of immunoblotting is shown in . (E,F) Transmission electron microscopy reveals that ODA and IDA were defective in Pih1d3-KO rats. Red arrows point to ODA and green arrows point to IDA in WT rats, whereas red arrowheads point to the absence of ODA and green arrowheads point to the absence of IDA in Pih1d3-KO rats. Tracheas dissected from KO and WT rats aged 20 days were examined by transmission electron microscopy. Scale bar: 20 nm. (G,H) Diagrams showing the structure of motile cilia. Compared to WT rats, ODA and IDA were defected in Pih1d3-KO rats.

Journal: Frontiers in Cell and Developmental Biology

Article Title: PIH1D3-knockout rats exhibit full ciliopathy features and dysfunctional pre-assembly and loading of dynein arms in motile cilia

doi: 10.3389/fcell.2023.1282787

Figure Lengend Snippet: Defective ODA and IDA in the motile cilia of PIH1D3-KO rats. (A–C) Immunofluorescent staining reveals that PIH1D3 deficiency in KO rats results in the dislocation of DNAI1 and DNALI1 from motile cilia labeled with acetyl-TUBA1A. The efferent ductules were dissected from Pih1d3-KO rats and WT littermates at the age of 27 days. Scale bar: 30 µm. (D) Immunoblotting shows that the expression of the ODA markers DNAI1 and DNAI2 and the IDA marker DNALI1 was reduced in the tissues of Pih1d3-KO rats compared to WT littermates. Rats of age postnatal day 20 were used for dissecting the trachea and testis, and rats of age postnatal day 0 were used for dissecting brains. Equal loading of 20 µg total protein per lane was verified by immunoblotting for actin. Quantification for the density of immunoblotting is shown in . (E,F) Transmission electron microscopy reveals that ODA and IDA were defective in Pih1d3-KO rats. Red arrows point to ODA and green arrows point to IDA in WT rats, whereas red arrowheads point to the absence of ODA and green arrowheads point to the absence of IDA in Pih1d3-KO rats. Tracheas dissected from KO and WT rats aged 20 days were examined by transmission electron microscopy. Scale bar: 20 nm. (G,H) Diagrams showing the structure of motile cilia. Compared to WT rats, ODA and IDA were defected in Pih1d3-KO rats.

Article Snippet: The following primary antibodies were used: rabbit polyclonal anti-PIH1D3 (Proteintech, 25309-1-AP); mouse monoclonal anti-acetylated alpha-tubulin IgG1 (Proteintech, CL488-66200); rabbit polyclonal anti-DNAI1 (Abcam, ab171964); rabbit polyclonal anti-DNAI2 (Proteintech, 17533-1-AP); rabbit polyclonal anti-DNALI1 for immunofluorescence staining (Proteintech, 17601-1-AP); mouse monoclonal anti-DNALI1 for immunoblotting (Santa Cruz, Sc-514831); rabbit polyclonal anti-IFT52 (Proteintech, 17534-1-AP); rabbit polyclonal anti-IFT57 (Proteintech, 11083-1-AP); rabbit polyclonal anti-PIH1D1 (Proteintech, 19427-1-AP); rabbit polyclonal anti-IFT20 (Proteintech, 13615-1-AP); rabbit polyclonal anti-IFT81 (Proteintech, 11744-1-AP); mouse monoclonal anti-Flag (Novus, NBP1-97410); and mouse monoclonal anti-Myc (Invitrogen, MA1-21316).

Techniques: Staining, Labeling, Western Blot, Expressing, Marker, Transmission Assay, Electron Microscopy

PIH1D3 protein partners revealed by immunoprecipitation (IP) combined with immunoblotting (IB). (A) Schematics showing the construction of plasmids expressing Myc-tagged partner proteins and flag-tagged full-length and fragmented PIH1D3 peptides. (B) Plasmid expressing that an individual partner protein was co-transfected with a PIH1D3-expressing plasmid into HEK293 cells, and the cell lysates were analyzed by IP and IB 48 h after transfection. (C) Plasmids expressing full-length or fragmented PIH1D3 peptides were co-transfected individually with a partner-expressing plasmid into HEK293 cells. The binding affinity of each PIH1D3 peptide for partner proteins was assessed by IP and IB. (D) Schematics showing the approximate binding sites on PIH1D3 for each partner protein examined.

Journal: Frontiers in Cell and Developmental Biology

Article Title: PIH1D3-knockout rats exhibit full ciliopathy features and dysfunctional pre-assembly and loading of dynein arms in motile cilia

doi: 10.3389/fcell.2023.1282787

Figure Lengend Snippet: PIH1D3 protein partners revealed by immunoprecipitation (IP) combined with immunoblotting (IB). (A) Schematics showing the construction of plasmids expressing Myc-tagged partner proteins and flag-tagged full-length and fragmented PIH1D3 peptides. (B) Plasmid expressing that an individual partner protein was co-transfected with a PIH1D3-expressing plasmid into HEK293 cells, and the cell lysates were analyzed by IP and IB 48 h after transfection. (C) Plasmids expressing full-length or fragmented PIH1D3 peptides were co-transfected individually with a partner-expressing plasmid into HEK293 cells. The binding affinity of each PIH1D3 peptide for partner proteins was assessed by IP and IB. (D) Schematics showing the approximate binding sites on PIH1D3 for each partner protein examined.

Article Snippet: The following primary antibodies were used: rabbit polyclonal anti-PIH1D3 (Proteintech, 25309-1-AP); mouse monoclonal anti-acetylated alpha-tubulin IgG1 (Proteintech, CL488-66200); rabbit polyclonal anti-DNAI1 (Abcam, ab171964); rabbit polyclonal anti-DNAI2 (Proteintech, 17533-1-AP); rabbit polyclonal anti-DNALI1 for immunofluorescence staining (Proteintech, 17601-1-AP); mouse monoclonal anti-DNALI1 for immunoblotting (Santa Cruz, Sc-514831); rabbit polyclonal anti-IFT52 (Proteintech, 17534-1-AP); rabbit polyclonal anti-IFT57 (Proteintech, 11083-1-AP); rabbit polyclonal anti-PIH1D1 (Proteintech, 19427-1-AP); rabbit polyclonal anti-IFT20 (Proteintech, 13615-1-AP); rabbit polyclonal anti-IFT81 (Proteintech, 11744-1-AP); mouse monoclonal anti-Flag (Novus, NBP1-97410); and mouse monoclonal anti-Myc (Invitrogen, MA1-21316).

Techniques: Immunoprecipitation, Western Blot, Expressing, Plasmid Preparation, Transfection, Binding Assay

Defected assembling and loading of ODA in KO rats. In WT rats, 1) PIH1D3 functions as a critical part of the protein complex mediating the pre-assembly and assembly of ODA through interacting with the DNAAF family and 2) PIH1D3 participates in the loading of the ODA complex onto vehicles for anterograde trafficking to ciliary tips by interacting with IFT52, IFT57, and others. In KO rats, 1) the processes for ODA assembly and loading are defected, leading to the degradation of ODA components such as DNAI1 and DNAI2; 2) ODA is absent in motile cilia; and 3) enhanced anterograde trafficking may occur partially due to compensatory mechanisms, leading to the accumulation of anterograde trafficking-associated proteins such as IFT52 and IFT20 in motile cilia. Similar to ODA, IDA is also defected when PIH1D3 is deleted in KO rats.

Journal: Frontiers in Cell and Developmental Biology

Article Title: PIH1D3-knockout rats exhibit full ciliopathy features and dysfunctional pre-assembly and loading of dynein arms in motile cilia

doi: 10.3389/fcell.2023.1282787

Figure Lengend Snippet: Defected assembling and loading of ODA in KO rats. In WT rats, 1) PIH1D3 functions as a critical part of the protein complex mediating the pre-assembly and assembly of ODA through interacting with the DNAAF family and 2) PIH1D3 participates in the loading of the ODA complex onto vehicles for anterograde trafficking to ciliary tips by interacting with IFT52, IFT57, and others. In KO rats, 1) the processes for ODA assembly and loading are defected, leading to the degradation of ODA components such as DNAI1 and DNAI2; 2) ODA is absent in motile cilia; and 3) enhanced anterograde trafficking may occur partially due to compensatory mechanisms, leading to the accumulation of anterograde trafficking-associated proteins such as IFT52 and IFT20 in motile cilia. Similar to ODA, IDA is also defected when PIH1D3 is deleted in KO rats.

Article Snippet: The following primary antibodies were used: rabbit polyclonal anti-PIH1D3 (Proteintech, 25309-1-AP); mouse monoclonal anti-acetylated alpha-tubulin IgG1 (Proteintech, CL488-66200); rabbit polyclonal anti-DNAI1 (Abcam, ab171964); rabbit polyclonal anti-DNAI2 (Proteintech, 17533-1-AP); rabbit polyclonal anti-DNALI1 for immunofluorescence staining (Proteintech, 17601-1-AP); mouse monoclonal anti-DNALI1 for immunoblotting (Santa Cruz, Sc-514831); rabbit polyclonal anti-IFT52 (Proteintech, 17534-1-AP); rabbit polyclonal anti-IFT57 (Proteintech, 11083-1-AP); rabbit polyclonal anti-PIH1D1 (Proteintech, 19427-1-AP); rabbit polyclonal anti-IFT20 (Proteintech, 13615-1-AP); rabbit polyclonal anti-IFT81 (Proteintech, 11744-1-AP); mouse monoclonal anti-Flag (Novus, NBP1-97410); and mouse monoclonal anti-Myc (Invitrogen, MA1-21316).

Techniques: