Structured Review

Millipore β gal
Abnormal holin lesions can discriminate between the endolysin and the <t>endolysin-β-Gal</t> hybrid. (A) The early-lysis allele of λ S supports lysis with both R and R φ lacZ . Culture growth and lysis were monitored by determining the A 550 after induction of λ Δ ( SR ) lysogens carrying pS105R − (circles) or pS105A52GR − (squares). The lysogens also carried a compatible plasmid carrying pZA32-mycR (open symbols) or pZA32-mycRφlacZ (filled symbols). IPTG (1 mM) was added at the time of lysogenic induction (time zero). (B and C) S holin lesions prematurely triggered by energy poisons are differentially permissive for the mycR and mycR-β-Gal endolysins. Culture growth and lysis were monitored by determining the A 550 after induction of the λ S105mycR (B) or λ S105mycR φ lacZ (C) lysogen. KCN (10 mM) was added at 20 min (open circles), 30 min (filled circles) or 40 min (open squares). Filled squares, untreated culture. (D) The parental T4 t , but not a t allele with its 69 C-terminal residues deleted, supports lysis with both cmycR and cmycR φ lacZ . Conditions were the same as described for panel A except that the plasmid was pT4-t (squares) or pT4-tΔ69 (circles).
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Images

1) Product Images from "Sizing the Holin Lesion with an Endolysin-?-Galactosidase Fusion"

Article Title: Sizing the Holin Lesion with an Endolysin-?-Galactosidase Fusion

Journal: Journal of Bacteriology

doi: 10.1128/JB.185.3.779-787.2003

Abnormal holin lesions can discriminate between the endolysin and the endolysin-β-Gal hybrid. (A) The early-lysis allele of λ S supports lysis with both R and R φ lacZ . Culture growth and lysis were monitored by determining the A 550 after induction of λ Δ ( SR ) lysogens carrying pS105R − (circles) or pS105A52GR − (squares). The lysogens also carried a compatible plasmid carrying pZA32-mycR (open symbols) or pZA32-mycRφlacZ (filled symbols). IPTG (1 mM) was added at the time of lysogenic induction (time zero). (B and C) S holin lesions prematurely triggered by energy poisons are differentially permissive for the mycR and mycR-β-Gal endolysins. Culture growth and lysis were monitored by determining the A 550 after induction of the λ S105mycR (B) or λ S105mycR φ lacZ (C) lysogen. KCN (10 mM) was added at 20 min (open circles), 30 min (filled circles) or 40 min (open squares). Filled squares, untreated culture. (D) The parental T4 t , but not a t allele with its 69 C-terminal residues deleted, supports lysis with both cmycR and cmycR φ lacZ . Conditions were the same as described for panel A except that the plasmid was pT4-t (squares) or pT4-tΔ69 (circles).
Figure Legend Snippet: Abnormal holin lesions can discriminate between the endolysin and the endolysin-β-Gal hybrid. (A) The early-lysis allele of λ S supports lysis with both R and R φ lacZ . Culture growth and lysis were monitored by determining the A 550 after induction of λ Δ ( SR ) lysogens carrying pS105R − (circles) or pS105A52GR − (squares). The lysogens also carried a compatible plasmid carrying pZA32-mycR (open symbols) or pZA32-mycRφlacZ (filled symbols). IPTG (1 mM) was added at the time of lysogenic induction (time zero). (B and C) S holin lesions prematurely triggered by energy poisons are differentially permissive for the mycR and mycR-β-Gal endolysins. Culture growth and lysis were monitored by determining the A 550 after induction of the λ S105mycR (B) or λ S105mycR φ lacZ (C) lysogen. KCN (10 mM) was added at 20 min (open circles), 30 min (filled circles) or 40 min (open squares). Filled squares, untreated culture. (D) The parental T4 t , but not a t allele with its 69 C-terminal residues deleted, supports lysis with both cmycR and cmycR φ lacZ . Conditions were the same as described for panel A except that the plasmid was pT4-t (squares) or pT4-tΔ69 (circles).

Techniques Used: Lysis, Plasmid Preparation

The endolysin-β-Gal fusion is fully functional in holin-mediated lysis. Culture growth and lysis were monitored by determining the A 550 after thermal inductions. (A) Induction of λ Δ ( SR ) lysogens carrying pS105mycR (circles) or pS105mycRφlacZ (squares). Open symbols, repeat experiment with approximately 1% CHCl 3 added at the indicated time; filled symbols, untreated cultures. (B) Induction of MC4100 lysogens carrying λ mycR or λ mycR φ lacZ prophages.
Figure Legend Snippet: The endolysin-β-Gal fusion is fully functional in holin-mediated lysis. Culture growth and lysis were monitored by determining the A 550 after thermal inductions. (A) Induction of λ Δ ( SR ) lysogens carrying pS105mycR (circles) or pS105mycRφlacZ (squares). Open symbols, repeat experiment with approximately 1% CHCl 3 added at the indicated time; filled symbols, untreated cultures. (B) Induction of MC4100 lysogens carrying λ mycR or λ mycR φ lacZ prophages.

Techniques Used: Functional Assay, Lysis

Modification of R with c- myc and lacZ at the 5′ and 3′ termini. Shown are the predicted alterations at the termini of the S and R proteins deriving from the insertions described in the text, in the context of the promoter-proximal region of the λ late transcriptional unit SRRzRz1 . The S allele is S105 ). The S ′ allele created by the insertion of the c- myc sequence at the start of R has 3 C-terminal residues altered, with a predicted net change of −2 in charge. The c-myc (10 residues) and β-Gal (1,021 residues) sequences are shown in underlined boldface as insertions after codon 1 and before codon 158 of R , respectively.
Figure Legend Snippet: Modification of R with c- myc and lacZ at the 5′ and 3′ termini. Shown are the predicted alterations at the termini of the S and R proteins deriving from the insertions described in the text, in the context of the promoter-proximal region of the λ late transcriptional unit SRRzRz1 . The S allele is S105 ). The S ′ allele created by the insertion of the c- myc sequence at the start of R has 3 C-terminal residues altered, with a predicted net change of −2 in charge. The c-myc (10 residues) and β-Gal (1,021 residues) sequences are shown in underlined boldface as insertions after codon 1 and before codon 158 of R , respectively.

Techniques Used: Modification, Sequencing

The lytic function of the hybrid mycR-β-Gal hybrid is not due to proteolytic release of the N-terminal endolysin domain. (A) Full-length mycR and mycR-β-Gal proteins accumulate to equivalent levels. Lysates from the induction of λ Δ ( SR ) lysogens carrying pS105mycR (lane 1) or pS105mycRφlacZ (lane 2) were analyzed by immunoblotting with anti-c- myc antibodies. Values at the left are molecular mass standards (in kilodaltons). (B) The level of mycR endolysin is limiting for the rate of lysis. Culture growth and lysis were monitored by determining the A 550 after the induction of λ Δ ( SR ) lysogens carrying pS105mycR (filled circles) or pS105mycRφlacZ (filled squares). Isogenic lysogens carrying pS105R − and the compatible plasmid pZS*32-mycR were subjected to lysogenic induction and the addition of either no IPTG (open circles) or 1 mM IPTG (open triangles) at time zero. (C) The levels of R-length proteolytic fragments of the mycR-β-Gal protein were negligible compared to lysis-limiting levels of MycR. Lysates from the induced lysogens carrying pS105mycRφlacZ or pS105R − and pZS*32-mycR (no IPTG) were analyzed as described for panel A, except that the stain was deliberately overdeveloped. Values at the right are molecular mass standards (in kilodaltons). (D) Gel filtration chromatography of the mycR and mycR-β-Gal proteins. A lysate prepared from induction of λ Δ ( SR )pS105mycRφlacZ was analyzed by gel filtration, followed by SDS-PAGE and immunoblotting with anti-c- myc antibodies. Shown is the elution pattern for mass standards as indicated, with the blot being superimposed under the appropriate samples. MAU, mass arbitrary units.
Figure Legend Snippet: The lytic function of the hybrid mycR-β-Gal hybrid is not due to proteolytic release of the N-terminal endolysin domain. (A) Full-length mycR and mycR-β-Gal proteins accumulate to equivalent levels. Lysates from the induction of λ Δ ( SR ) lysogens carrying pS105mycR (lane 1) or pS105mycRφlacZ (lane 2) were analyzed by immunoblotting with anti-c- myc antibodies. Values at the left are molecular mass standards (in kilodaltons). (B) The level of mycR endolysin is limiting for the rate of lysis. Culture growth and lysis were monitored by determining the A 550 after the induction of λ Δ ( SR ) lysogens carrying pS105mycR (filled circles) or pS105mycRφlacZ (filled squares). Isogenic lysogens carrying pS105R − and the compatible plasmid pZS*32-mycR were subjected to lysogenic induction and the addition of either no IPTG (open circles) or 1 mM IPTG (open triangles) at time zero. (C) The levels of R-length proteolytic fragments of the mycR-β-Gal protein were negligible compared to lysis-limiting levels of MycR. Lysates from the induced lysogens carrying pS105mycRφlacZ or pS105R − and pZS*32-mycR (no IPTG) were analyzed as described for panel A, except that the stain was deliberately overdeveloped. Values at the right are molecular mass standards (in kilodaltons). (D) Gel filtration chromatography of the mycR and mycR-β-Gal proteins. A lysate prepared from induction of λ Δ ( SR )pS105mycRφlacZ was analyzed by gel filtration, followed by SDS-PAGE and immunoblotting with anti-c- myc antibodies. Shown is the elution pattern for mass standards as indicated, with the blot being superimposed under the appropriate samples. MAU, mass arbitrary units.

Techniques Used: Lysis, Plasmid Preparation, Staining, Filtration, Chromatography, SDS Page

2) Product Images from "A recurrent WARS mutation is a novel cause of autosomal dominant distal hereditary motor neuropathy"

Article Title: A recurrent WARS mutation is a novel cause of autosomal dominant distal hereditary motor neuropathy

Journal: Brain

doi: 10.1093/brain/awx058

In vitro characterization of wild-type and His257Arg (H257R) mutant TrpRS proteins. ( A ) β-Gal reporter assay and ( B ) firefly luciferase reporter assay demonstrating that H257R TrpRS has a dominant-negative effect on protein synthesis. HEK293 cells co-transfected with β-Gal or firefly luciferase reporter plasmids, along with different ratios of wild-type and H257R TrpRS expression plasmids were lysed and assayed for β-Gal or firefly luciferase activities at 48 h after transfection. The error bars indicate standard errors of the mean ( n = 3) and asterisks indicate statistically significant differences (** P
Figure Legend Snippet: In vitro characterization of wild-type and His257Arg (H257R) mutant TrpRS proteins. ( A ) β-Gal reporter assay and ( B ) firefly luciferase reporter assay demonstrating that H257R TrpRS has a dominant-negative effect on protein synthesis. HEK293 cells co-transfected with β-Gal or firefly luciferase reporter plasmids, along with different ratios of wild-type and H257R TrpRS expression plasmids were lysed and assayed for β-Gal or firefly luciferase activities at 48 h after transfection. The error bars indicate standard errors of the mean ( n = 3) and asterisks indicate statistically significant differences (** P

Techniques Used: In Vitro, Mutagenesis, Reporter Assay, Luciferase, Dominant Negative Mutation, Transfection, Expressing

3) Product Images from "Plexin-B2 Regulates the Proliferation and Migration of Neuroblasts in the Postnatal and Adult Subventricular Zone"

Article Title: Plexin-B2 Regulates the Proliferation and Migration of Neuroblasts in the Postnatal and Adult Subventricular Zone

Journal: The Journal of Neuroscience

doi: 10.1523/JNEUROSCI.0344-12.2012

Plexin-B2 expression in the different cell populations of the adult SVZ and RMS. A is a coronal section of the forebrain at the level of the SVZ. SVZ cells lining the ventricle (V) strongly express Plexin-B2 (arrowheads). B–D , Rapidly proliferating type C cells labeled with a short-pulse of BrdU ( B ), phosphohistone 3 (H3P) ( C ), and Ki67 ( D ) are immunoreactive for Plexin-B2 (arrowheads). E–G , Plexin-B2 is also expressed by migrating neuroblasts labeled with PSA-NCAM in the SVZ ( E , F , arrowheads) and by DCX in the RMS ( G , arrowheads). H–K , In the SVZ, GFAP+ cells express Plexin-B2 ( H–J , arrowheads) but GFAP astrocytes ( K , arrows) ensheathing migrating neuroblasts (visualized by Hoechst staining) in the RMS do not express β-gal in Plxnb2 +/− mice. Scale bars: A , 100 μm; B , 12 μm; C , D , 7 μm; E , F , K , 15 μm; G , 25 μm; H–J , 14 μm.
Figure Legend Snippet: Plexin-B2 expression in the different cell populations of the adult SVZ and RMS. A is a coronal section of the forebrain at the level of the SVZ. SVZ cells lining the ventricle (V) strongly express Plexin-B2 (arrowheads). B–D , Rapidly proliferating type C cells labeled with a short-pulse of BrdU ( B ), phosphohistone 3 (H3P) ( C ), and Ki67 ( D ) are immunoreactive for Plexin-B2 (arrowheads). E–G , Plexin-B2 is also expressed by migrating neuroblasts labeled with PSA-NCAM in the SVZ ( E , F , arrowheads) and by DCX in the RMS ( G , arrowheads). H–K , In the SVZ, GFAP+ cells express Plexin-B2 ( H–J , arrowheads) but GFAP astrocytes ( K , arrows) ensheathing migrating neuroblasts (visualized by Hoechst staining) in the RMS do not express β-gal in Plxnb2 +/− mice. Scale bars: A , 100 μm; B , 12 μm; C , D , 7 μm; E , F , K , 15 μm; G , 25 μm; H–J , 14 μm.

Techniques Used: Expressing, Labeling, Staining, Mouse Assay

Plexin-B2 is expressed by tangentially migrating neuroblasts and by PG cells. A–D , Sagittal sections at the level of the OB of a P7 mice electroporated at P2 in the SVZ with a GFP-plasmid and immunostained with an anti-Plexin-B2 antibody. Plexin-B2 is highly expressed in the RMS containing tangentially migrating GFP+ neuroblasts. C , C′ is an example of migrating neuroblasts expressing GFP in the cytosol and Plexin-B2 at the cell surface (asterisks). The arrowheads point to the tip of labeled leading processes. Plexin-B2 expression is downregulated in the granular cell layer (GCL), which contains radially migrating GFP+ cells ( D , arrowhead). E , F , Sagittal section of the adult OB immunostained with anti-Plexin-B2 and counterstained with Hoechst. Plexin-B2 is detected in the RMS, in the mitral cell layer (MCL), and in the glomerular cell layer (GL), but not in the granule cell layer (GCL). F shows that adult mitral cells are immunoreactive for Plexin-B2. G–L are sections of the adult OB of Plxnb2 +/− ( G–J ) or wild-type mice, at the level of the glomerular layer. In Plxnb2 +/− ( G–J ) OB, β-galactosidase is detected in all the different subtypes of periglomerular cells expressing Pax6 ( G , arrow), GABA ( H , arrow), calretinin ( I , arrow), and TH ( J , arrow). Likewise, TH-positive PG cells are immunoreactive for Plexin-B2 ( L , arrow). In all panels, the arrowheads indicate cells that express β-gal or Plexin-B2 and are not labeled with the other markers. Scale bars: A , B , 160 μm; C , C′ , 22 μm; D , 34 μm; E , 70 μm; F , 30 μm; K , L , 20 μm.
Figure Legend Snippet: Plexin-B2 is expressed by tangentially migrating neuroblasts and by PG cells. A–D , Sagittal sections at the level of the OB of a P7 mice electroporated at P2 in the SVZ with a GFP-plasmid and immunostained with an anti-Plexin-B2 antibody. Plexin-B2 is highly expressed in the RMS containing tangentially migrating GFP+ neuroblasts. C , C′ is an example of migrating neuroblasts expressing GFP in the cytosol and Plexin-B2 at the cell surface (asterisks). The arrowheads point to the tip of labeled leading processes. Plexin-B2 expression is downregulated in the granular cell layer (GCL), which contains radially migrating GFP+ cells ( D , arrowhead). E , F , Sagittal section of the adult OB immunostained with anti-Plexin-B2 and counterstained with Hoechst. Plexin-B2 is detected in the RMS, in the mitral cell layer (MCL), and in the glomerular cell layer (GL), but not in the granule cell layer (GCL). F shows that adult mitral cells are immunoreactive for Plexin-B2. G–L are sections of the adult OB of Plxnb2 +/− ( G–J ) or wild-type mice, at the level of the glomerular layer. In Plxnb2 +/− ( G–J ) OB, β-galactosidase is detected in all the different subtypes of periglomerular cells expressing Pax6 ( G , arrow), GABA ( H , arrow), calretinin ( I , arrow), and TH ( J , arrow). Likewise, TH-positive PG cells are immunoreactive for Plexin-B2 ( L , arrow). In all panels, the arrowheads indicate cells that express β-gal or Plexin-B2 and are not labeled with the other markers. Scale bars: A , B , 160 μm; C , C′ , 22 μm; D , 34 μm; E , 70 μm; F , 30 μm; K , L , 20 μm.

Techniques Used: Mouse Assay, Plasmid Preparation, Expressing, Labeling

Plexin-B2 is expressed in the postnatal and adult SVZ–RMS. Sections A–F are coronal. A–C , In the adult, in situ hybridization with Plxnb2 riboprobe showed that Plxnb2 is highly expressed in the RMS ( A , B , arrowhead), in the glomerular layer (gl) ( A , arrows), and in the subventricular zone ( C , arrowhead). The insets in A and C are high magnification of the boxed area. D–F , β-Galactosidase immunohistochemistry also revealed similar expression patterns in all three regions (insets are high magnification of the boxed areas). G , Sagittal section of the telencephalon of P8 Plxnb2 +/− mouse labeled for PLAP. PLAP is highly expressed in the RMS (arrowhead) and some olfactory axons (arrow). H , Sagittal section of the telencephalon in a P21 mouse immunolabeled with anti-Plexin-B2 antibodies. Plexin-B2 is highly expressed in the RMS (arrowhead) and some olfactory axons (arrow). Scale bars: A–E , 600 μm; F , 200 μm; G , 500 μm; H , 600 μm.
Figure Legend Snippet: Plexin-B2 is expressed in the postnatal and adult SVZ–RMS. Sections A–F are coronal. A–C , In the adult, in situ hybridization with Plxnb2 riboprobe showed that Plxnb2 is highly expressed in the RMS ( A , B , arrowhead), in the glomerular layer (gl) ( A , arrows), and in the subventricular zone ( C , arrowhead). The insets in A and C are high magnification of the boxed area. D–F , β-Galactosidase immunohistochemistry also revealed similar expression patterns in all three regions (insets are high magnification of the boxed areas). G , Sagittal section of the telencephalon of P8 Plxnb2 +/− mouse labeled for PLAP. PLAP is highly expressed in the RMS (arrowhead) and some olfactory axons (arrow). H , Sagittal section of the telencephalon in a P21 mouse immunolabeled with anti-Plexin-B2 antibodies. Plexin-B2 is highly expressed in the RMS (arrowhead) and some olfactory axons (arrow). Scale bars: A–E , 600 μm; F , 200 μm; G , 500 μm; H , 600 μm.

Techniques Used: In Situ Hybridization, Immunohistochemistry, Expressing, Labeling, Immunolabeling

4) Product Images from "Mice lacking mitochondrial ferritin are more sensitive to doxorubicin-mediated cardiotoxicity"

Article Title: Mice lacking mitochondrial ferritin are more sensitive to doxorubicin-mediated cardiotoxicity

Journal: Journal of Molecular Medicine (Berlin, Germany)

doi: 10.1007/s00109-014-1147-0

Construction and genotyping of FtMt −/− mice. a Schematic of the FtMt locus and targeting vector, the arrows indicate the PCR primers for genotyping. b Genotyping gel of mice wild type (+/+), heterozygous (+/−), and homozygous (−/−) for the inactivated FtMt allele. In the M lane, a DNA molecular weight ladder was loaded. The size of the expected PCR amplicons is indicated. c RT-PCR analysis of testis RNA extracted from wild type (+/+), heterozygous (+/−), and homozygous (−/−) mice and amplified with primers for FtMt, beta-galactosidase (β-gal, encoded by the Lac-Z gene), and the housekeeping transcript HPRT1. d Immunoblotting of protein extracts from the testis of wild type (+/+), heterozygous (+/−), and homozygous (−/−) mice overlaid with antibodies for mouse FtMt, β-gal, and GAPDH as loading calibrator. e Immunoblotting of protein extracts from various tissues of wild type (+/+) and homozygous (−/−) mice, overlaid with antibodies for mouse FtMt. ( f , g ) Immunohistochemical detection of FtMt in the testis of wild type ( f ) and FtMt(−/−) ( g ) mice. Bar 50 μm
Figure Legend Snippet: Construction and genotyping of FtMt −/− mice. a Schematic of the FtMt locus and targeting vector, the arrows indicate the PCR primers for genotyping. b Genotyping gel of mice wild type (+/+), heterozygous (+/−), and homozygous (−/−) for the inactivated FtMt allele. In the M lane, a DNA molecular weight ladder was loaded. The size of the expected PCR amplicons is indicated. c RT-PCR analysis of testis RNA extracted from wild type (+/+), heterozygous (+/−), and homozygous (−/−) mice and amplified with primers for FtMt, beta-galactosidase (β-gal, encoded by the Lac-Z gene), and the housekeeping transcript HPRT1. d Immunoblotting of protein extracts from the testis of wild type (+/+), heterozygous (+/−), and homozygous (−/−) mice overlaid with antibodies for mouse FtMt, β-gal, and GAPDH as loading calibrator. e Immunoblotting of protein extracts from various tissues of wild type (+/+) and homozygous (−/−) mice, overlaid with antibodies for mouse FtMt. ( f , g ) Immunohistochemical detection of FtMt in the testis of wild type ( f ) and FtMt(−/−) ( g ) mice. Bar 50 μm

Techniques Used: Mouse Assay, Plasmid Preparation, Polymerase Chain Reaction, Molecular Weight, Reverse Transcription Polymerase Chain Reaction, Amplification, Immunohistochemistry

5) Product Images from "Fission of pancreatic islets during postnatal growth of the mouse"

Article Title: Fission of pancreatic islets during postnatal growth of the mouse

Journal: Journal of Anatomy

doi: 10.1111/j.1469-7580.2004.00265.x

Cell composition of α- and β-cell populations. Islets from H253 female hemizygotes are double-labelled for β-gal (fluorescein) and the appropriate hormone (TRITC/Texas Red). Insulin + β-cell populations and glucagon + α-cell populations are both of heterogeneous β-gal + /β-gal − cell composition. (A) Glucagon. (B) Insulin. Scale bar = 50 µm.
Figure Legend Snippet: Cell composition of α- and β-cell populations. Islets from H253 female hemizygotes are double-labelled for β-gal (fluorescein) and the appropriate hormone (TRITC/Texas Red). Insulin + β-cell populations and glucagon + α-cell populations are both of heterogeneous β-gal + /β-gal − cell composition. (A) Glucagon. (B) Insulin. Scale bar = 50 µm.

Techniques Used:

Analysis of cell composition of dumb-bell and non-dumb-bell islets. (A,B) Relationship between percentage β-gal + cells and edge-to-edge distance in random pairs of non-dumb-bell islets. (A) Including outliers (asterisks) ( n = 65 pairs). (B) Outliers excluded ( n = 63 pairs). (C) Relationship between percentages of β-gal + cells on the two sides of each of 23 dumb-bell islets.
Figure Legend Snippet: Analysis of cell composition of dumb-bell and non-dumb-bell islets. (A,B) Relationship between percentage β-gal + cells and edge-to-edge distance in random pairs of non-dumb-bell islets. (A) Including outliers (asterisks) ( n = 65 pairs). (B) Outliers excluded ( n = 63 pairs). (C) Relationship between percentages of β-gal + cells on the two sides of each of 23 dumb-bell islets.

Techniques Used:

‘Dumbbell’ islets in histological sections. (A,B) Paraffin sections of X-gal-stained mosaic female pancreas immunoperoxidase-stained for glucagon. (C) Frozen section of mosaic female pancreas immunofluorescence-stained for β-gal (fluorescein) and glucagon (Texas Red). Note neck of glucagon + cells (arrows). Scale bar = 100 µm.
Figure Legend Snippet: ‘Dumbbell’ islets in histological sections. (A,B) Paraffin sections of X-gal-stained mosaic female pancreas immunoperoxidase-stained for glucagon. (C) Frozen section of mosaic female pancreas immunofluorescence-stained for β-gal (fluorescein) and glucagon (Texas Red). Note neck of glucagon + cells (arrows). Scale bar = 100 µm.

Techniques Used: Staining, Immunofluorescence

Neonatal islets are of a heterogeneous cell composition but some undergo a conversion to a homogeneous state by 1 month after birth. (A) Heterogeneous islets in a PN7 mosaic female. Note fine-grained mosaicism and widespread intermingling of β-gal + and β-gal − cells. (B) Homogeneous β-gal + islets in a PN21 mosaic female. (C) Homogeneous β-gal − islets in a PN26 weeks mosaic female. Specimens stained with X-gal and immunoperoxidase-stained for glucagon. Scale bar = 100 µm.
Figure Legend Snippet: Neonatal islets are of a heterogeneous cell composition but some undergo a conversion to a homogeneous state by 1 month after birth. (A) Heterogeneous islets in a PN7 mosaic female. Note fine-grained mosaicism and widespread intermingling of β-gal + and β-gal − cells. (B) Homogeneous β-gal + islets in a PN21 mosaic female. (C) Homogeneous β-gal − islets in a PN26 weeks mosaic female. Specimens stained with X-gal and immunoperoxidase-stained for glucagon. Scale bar = 100 µm.

Techniques Used: Staining

Characteristic pattern of X-gal staining for β-gal activity in the intestinal crypts. (A) H253 homozygous female. (B) Hemizygous, X-inactivation mosaic female. Scale bar = 100 µm.
Figure Legend Snippet: Characteristic pattern of X-gal staining for β-gal activity in the intestinal crypts. (A) H253 homozygous female. (B) Hemizygous, X-inactivation mosaic female. Scale bar = 100 µm.

Techniques Used: Staining, Activity Assay

β-gal activity is retained in the islets but declines in the acini. (A) X-gal staining is absent in some acini in a PN7 transgenic male. (B) Complete loss of acinar β-gal activity in a PN12 weeks homozygous female. Specimens stained with X-gal and immunoperoxidase-stained for glucagon. Scale bar = 100 µm.
Figure Legend Snippet: β-gal activity is retained in the islets but declines in the acini. (A) X-gal staining is absent in some acini in a PN7 transgenic male. (B) Complete loss of acinar β-gal activity in a PN12 weeks homozygous female. Specimens stained with X-gal and immunoperoxidase-stained for glucagon. Scale bar = 100 µm.

Techniques Used: Activity Assay, Staining, Transgenic Assay

6) Product Images from "Performance of Genomic Bordering Elements at Predefined Genomic Loci"

Article Title: Performance of Genomic Bordering Elements at Predefined Genomic Loci

Journal: Molecular and Cellular Biology

doi: 10.1128/MCB.25.6.2260-2272.2005

Level of β-Gal expression in five parental clones established for RMCE. The parental construct containing a hyg-tk fusion gene under the control of a tk promoter (pTK) is exchanged for a lacZ-neo fusion gene cassette that resides on the exchange
Figure Legend Snippet: Level of β-Gal expression in five parental clones established for RMCE. The parental construct containing a hyg-tk fusion gene under the control of a tk promoter (pTK) is exchanged for a lacZ-neo fusion gene cassette that resides on the exchange

Techniques Used: Expressing, Clone Assay, Construct

7) Product Images from "p27 Protein Protects Metabolically Stressed Cardiomyocytes from Apoptosis by Promoting Autophagy *"

Article Title: p27 Protein Protects Metabolically Stressed Cardiomyocytes from Apoptosis by Promoting Autophagy *

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.M113.542795

TAT-p27-reduced cardiomyocyte apoptosis following glucose deprivation is autophagy-dependent. Glucose-deprived ( GD ) cardiomyocytes were treated with TAT-p27 in the presence or absence of autophagy flux inhibitor Baf-A1. A and B , TAT-p27-treated glucose-deprived cardiomyocytes showed increased LC3-II levels and reduced p62 and caspase 3 cleavage compared with TAT-β-Gal ( lanes 3 versus 1 ). However, addition of Baf-A1 increased apoptosis. *, p
Figure Legend Snippet: TAT-p27-reduced cardiomyocyte apoptosis following glucose deprivation is autophagy-dependent. Glucose-deprived ( GD ) cardiomyocytes were treated with TAT-p27 in the presence or absence of autophagy flux inhibitor Baf-A1. A and B , TAT-p27-treated glucose-deprived cardiomyocytes showed increased LC3-II levels and reduced p62 and caspase 3 cleavage compared with TAT-β-Gal ( lanes 3 versus 1 ). However, addition of Baf-A1 increased apoptosis. *, p

Techniques Used:

TAT-p27 reduced apoptosis post-MI by promoting autophagy. A and B , homogenates from mice undergoing sham and LAD ligation surgery, with the latter receiving treatments with TAT-p27 or TAT-β-Gal alone or in combination with the autophagy inhibitor CQ, were probed for LC3-II, p62, and cleaved caspase 3. β-Tubulin served as loading control. Compared with sham-operated controls, post-MI groups revealed higher levels of LC3-II and cleaved caspase 3 levels (*, p
Figure Legend Snippet: TAT-p27 reduced apoptosis post-MI by promoting autophagy. A and B , homogenates from mice undergoing sham and LAD ligation surgery, with the latter receiving treatments with TAT-p27 or TAT-β-Gal alone or in combination with the autophagy inhibitor CQ, were probed for LC3-II, p62, and cleaved caspase 3. β-Tubulin served as loading control. Compared with sham-operated controls, post-MI groups revealed higher levels of LC3-II and cleaved caspase 3 levels (*, p

Techniques Used: Mouse Assay, Ligation

TAT-p27 reduced infarct size, prevented adverse cardiac remodeling, and improved cardiac function post-MI through enhanced autophagy. A , experimental scheme. B , fractional shortening ( FS ; percentage), LVISD and LVIDD (mm) were determined by M-mode echocardiography 14 days after sham or LAD ligation surgery, with the latter groups receiving treatment with TAT-β-Gal or TAT-p27 in the presence or absence of the autophagy inhibitor CQ. TAT-p27 treatment limited the reduced fractional shortening and increased LVISD and LVIDD observed post-MI in TAT-β-Gal-treated controls. This effect of TAT-p27 was lost in the presence of CQ. C , cardiac sections stained for determination of infarct area. The ability of TAT-p27 to reduce infarct size was also abolished in the presence of CQ. Both analyses have n = 5–7/group; *, p
Figure Legend Snippet: TAT-p27 reduced infarct size, prevented adverse cardiac remodeling, and improved cardiac function post-MI through enhanced autophagy. A , experimental scheme. B , fractional shortening ( FS ; percentage), LVISD and LVIDD (mm) were determined by M-mode echocardiography 14 days after sham or LAD ligation surgery, with the latter groups receiving treatment with TAT-β-Gal or TAT-p27 in the presence or absence of the autophagy inhibitor CQ. TAT-p27 treatment limited the reduced fractional shortening and increased LVISD and LVIDD observed post-MI in TAT-β-Gal-treated controls. This effect of TAT-p27 was lost in the presence of CQ. C , cardiac sections stained for determination of infarct area. The ability of TAT-p27 to reduce infarct size was also abolished in the presence of CQ. Both analyses have n = 5–7/group; *, p

Techniques Used: Ligation, Staining

8) Product Images from "Promotion of Dendritic Growth by CPG15, an Activity-Induced Signaling Molecule"

Article Title: Promotion of Dendritic Growth by CPG15, an Activity-Induced Signaling Molecule

Journal: Science (New York, N.Y.)

doi:

Quantification of CPG15 growth-promoting effect on dendritic arbors. ( A ) The average TDBL of rostrally projecting CPG15 neurons, β-gal neurons, and uninfected control neurons is plotted over 3 days of imaging. On the first day of imaging, the average TDBL of projection neurons from CPG15VV-infected animals was 447 ± 69 μm ( n = 39), significantly larger ( P
Figure Legend Snippet: Quantification of CPG15 growth-promoting effect on dendritic arbors. ( A ) The average TDBL of rostrally projecting CPG15 neurons, β-gal neurons, and uninfected control neurons is plotted over 3 days of imaging. On the first day of imaging, the average TDBL of projection neurons from CPG15VV-infected animals was 447 ± 69 μm ( n = 39), significantly larger ( P

Techniques Used: Imaging, Infection

CPG15 induction by kainic acid and its expression in Xenopus optic tectum. ( A ) Immuno-blot of protein extracts from tadpoles harvested at the specified times after intraventricular injection of KA, or rat hippocampus dentate gyri 24 hours after ip injection of KA (right lane). Incubation with the antiserum to CPG15 labels a 12-kD band (arrow) that is not seen with preimmune serum (PI). Confocal images of sections through the optic tecti of untreated tadpoles ( B and C ) or tadpoles infected with CPG15VV ( D and E ) or CPG15t3VV ( F and G ). Sections probed with preimmune rabbit serum show no specific labeling (B). Outlined on this section are the optic tectal neuropil (N), differentiated tectal neurons (TN), and the proliferative zone (PZ). These same regions can be discerned in the sections stained with the antisera to CPG15 [(C), (E), and (G)]. A honeycomb pattern of endogenous CPG15 immunoreactivity can be seen in the TN region of the tectum, and retinal ganglion cell axons are stained in N (C). Sections from animals infected with virus were double-labeled with anti– β-gal to show extent of infection [(D) and (F)] and with anti-CPG15 at higher magnification [(E) and (G)]. In the infected tecti [(E) and (G)], the honeycomb pattern of CPG15 immunoreactivity also extends into the PZ, where many infected neurons are located [(D) and (F)]. Arrows mark retinotectal axons. Bar, 100 μm for upper panel and 50 μm for lower panels.
Figure Legend Snippet: CPG15 induction by kainic acid and its expression in Xenopus optic tectum. ( A ) Immuno-blot of protein extracts from tadpoles harvested at the specified times after intraventricular injection of KA, or rat hippocampus dentate gyri 24 hours after ip injection of KA (right lane). Incubation with the antiserum to CPG15 labels a 12-kD band (arrow) that is not seen with preimmune serum (PI). Confocal images of sections through the optic tecti of untreated tadpoles ( B and C ) or tadpoles infected with CPG15VV ( D and E ) or CPG15t3VV ( F and G ). Sections probed with preimmune rabbit serum show no specific labeling (B). Outlined on this section are the optic tectal neuropil (N), differentiated tectal neurons (TN), and the proliferative zone (PZ). These same regions can be discerned in the sections stained with the antisera to CPG15 [(C), (E), and (G)]. A honeycomb pattern of endogenous CPG15 immunoreactivity can be seen in the TN region of the tectum, and retinal ganglion cell axons are stained in N (C). Sections from animals infected with virus were double-labeled with anti– β-gal to show extent of infection [(D) and (F)] and with anti-CPG15 at higher magnification [(E) and (G)]. In the infected tecti [(E) and (G)], the honeycomb pattern of CPG15 immunoreactivity also extends into the PZ, where many infected neurons are located [(D) and (F)]. Arrows mark retinotectal axons. Bar, 100 μm for upper panel and 50 μm for lower panels.

Techniques Used: Expressing, Injection, Incubation, Infection, Labeling, Staining

CPG15 does not affect tectal interneurons. ( A ) Drawings of interneurons from β-gal– infected animals (left) and CPG15VV-infected animals (right) with a TDBL closest to the mean branch length of each group. ( B ) The growth rate (TDBL on day 2 – TDBL on day 1) is significantly greater ( P
Figure Legend Snippet: CPG15 does not affect tectal interneurons. ( A ) Drawings of interneurons from β-gal– infected animals (left) and CPG15VV-infected animals (right) with a TDBL closest to the mean branch length of each group. ( B ) The growth rate (TDBL on day 2 – TDBL on day 1) is significantly greater ( P

Techniques Used: Infection

CPG15 promotes growth through intercellular signaling. ( A ) A 3D reconstruction of a tectal projection neuron from a CPG15VV-infected animal, with a TDBL of 1684 μm on the first day of imaging (day 1) and 2021 μm on the second day (day 2). This cell is a clear outlier on both days as the largest control cell is 642 μm on the first day of imaging and 1017 μm on the second. ( B ) Top panel shows a drawing of the tadpole optic tectum (OT) and the tectal ventricle (V) with the marked location of this cell. The green square delineates the region shown in the bottom panel. Bottom panel shows a superimposition of images collected with a 488-nm filter visualizing the DiI-labeled cell imaged in green and images collected with a 647-nm filter visualizing β-gal immunopositive cells in red. The arrow marks the cell imaged in (A).
Figure Legend Snippet: CPG15 promotes growth through intercellular signaling. ( A ) A 3D reconstruction of a tectal projection neuron from a CPG15VV-infected animal, with a TDBL of 1684 μm on the first day of imaging (day 1) and 2021 μm on the second day (day 2). This cell is a clear outlier on both days as the largest control cell is 642 μm on the first day of imaging and 1017 μm on the second. ( B ) Top panel shows a drawing of the tadpole optic tectum (OT) and the tectal ventricle (V) with the marked location of this cell. The green square delineates the region shown in the bottom panel. Bottom panel shows a superimposition of images collected with a 488-nm filter visualizing the DiI-labeled cell imaged in green and images collected with a 647-nm filter visualizing β-gal immunopositive cells in red. The arrow marks the cell imaged in (A).

Techniques Used: Infection, Imaging, Labeling

9) Product Images from "Advantages of Hydrogel-Based 3D-Printed Enzyme Reactors and Their Limitations for Biocatalysis"

Article Title: Advantages of Hydrogel-Based 3D-Printed Enzyme Reactors and Their Limitations for Biocatalysis

Journal: Frontiers in Bioengineering and Biotechnology

doi: 10.3389/fbioe.2018.00211

Michaelis–Menten kinetics of the investigated biocatalytic reactions with freely dissolved enzymes. Kinetic parameters v max and K m were calculated by fitting the data to the Michaelis–Menten function, which is shown including 95% confidence bounds. In the whole figure, each data point represents one sample. (A) β-Gal kinetics for the cleavage of ONPG at pH 4.6 ( n = 3 separate runs). (B) BFD kinetics for the carboligation of benzaldehyde and a respective 2.5-fold excess of acetaldehyde ( n = 2 separate runs). Estimation of the confidence bounds was omitted because of limited data for substrate excess. (C) ADH kinetics for the reduction of acetophenone. (D) ADH kinetics for the reduction of ( S )-HPP, which was previously synthesized by BFD. For ADH kinetics, data points were generated in one batch due to shortage of the enzyme.
Figure Legend Snippet: Michaelis–Menten kinetics of the investigated biocatalytic reactions with freely dissolved enzymes. Kinetic parameters v max and K m were calculated by fitting the data to the Michaelis–Menten function, which is shown including 95% confidence bounds. In the whole figure, each data point represents one sample. (A) β-Gal kinetics for the cleavage of ONPG at pH 4.6 ( n = 3 separate runs). (B) BFD kinetics for the carboligation of benzaldehyde and a respective 2.5-fold excess of acetaldehyde ( n = 2 separate runs). Estimation of the confidence bounds was omitted because of limited data for substrate excess. (C) ADH kinetics for the reduction of acetophenone. (D) ADH kinetics for the reduction of ( S )-HPP, which was previously synthesized by BFD. For ADH kinetics, data points were generated in one batch due to shortage of the enzyme.

Techniques Used: Synthesized, Generated

Conversion curves in batch with enzymes entrapped in hydrogel lattices by following the formation of the respective product over time. (A) β-Gal kinetics for the cleavage of 2.2 mM ONPG at pH 4.6 ( n = 3 separate runs). (B) Formation of ( S )-HPP by BFD starting from 35 mM benzaldehyde and 87.5 mM acetaldehyde ( n = 2 separate runs). (C) ADH-catalyzed reduction of three different concentrations of acetophenone ( n = 2 separate runs). (D) ADH-catalyzed reduction of three different concentrations of ( S )-HPP, provided by BFD reactor experiments ( n = 1). Linear regression with a 95% confidence interval of the slope was applied.
Figure Legend Snippet: Conversion curves in batch with enzymes entrapped in hydrogel lattices by following the formation of the respective product over time. (A) β-Gal kinetics for the cleavage of 2.2 mM ONPG at pH 4.6 ( n = 3 separate runs). (B) Formation of ( S )-HPP by BFD starting from 35 mM benzaldehyde and 87.5 mM acetaldehyde ( n = 2 separate runs). (C) ADH-catalyzed reduction of three different concentrations of acetophenone ( n = 2 separate runs). (D) ADH-catalyzed reduction of three different concentrations of ( S )-HPP, provided by BFD reactor experiments ( n = 1). Linear regression with a 95% confidence interval of the slope was applied.

Techniques Used:

Detected conversion to the respective product compound at the exit of the 3 ml reactor system of one exemplary run. Parameters were (A) β-Gal ( m = 0.38 mg/reactor) with ONPG substrate 2.2 mM, flow rate 0.05 ml/min. (B) BFD ( m = 56 mg/reactor) catalyzing the carboligation of benzaldehyde (25 mM) and acetaldehyde (62.5 mM), flow rate 0.017 ml/min. (C) ADH ( m = 42 mg/reactor) reducing acetophenone (50 mM), flow rate 0.017 ml/min. Reproducibility is shown by a second experiment with identical parameters. (D) ADH ( m = 58 mg/reactor) reducing ( S )-HPP provided by BFD reactor experiments (11 mM), flow rate 0.017 ml/min.
Figure Legend Snippet: Detected conversion to the respective product compound at the exit of the 3 ml reactor system of one exemplary run. Parameters were (A) β-Gal ( m = 0.38 mg/reactor) with ONPG substrate 2.2 mM, flow rate 0.05 ml/min. (B) BFD ( m = 56 mg/reactor) catalyzing the carboligation of benzaldehyde (25 mM) and acetaldehyde (62.5 mM), flow rate 0.017 ml/min. (C) ADH ( m = 42 mg/reactor) reducing acetophenone (50 mM), flow rate 0.017 ml/min. Reproducibility is shown by a second experiment with identical parameters. (D) ADH ( m = 58 mg/reactor) reducing ( S )-HPP provided by BFD reactor experiments (11 mM), flow rate 0.017 ml/min.

Techniques Used: Flow Cytometry

10) Product Images from "ygiW and qseBC are co-expressed in Aggregatibacter actinomycetemcomitans and regulate biofilm growth"

Article Title: ygiW and qseBC are co-expressed in Aggregatibacter actinomycetemcomitans and regulate biofilm growth

Journal: Microbiology

doi: 10.1099/mic.0.066183-0

β-gal activity of wild-type A. actinomycetemcomitans 652 and isogenic ifΔ ygiW, Δ qseB , Δ qseBC and qseC Δpr mutants harbouring pDJR29. (a) Graphic representation of the transcriptional reporter plasmids pDJR29. (b)
Figure Legend Snippet: β-gal activity of wild-type A. actinomycetemcomitans 652 and isogenic ifΔ ygiW, Δ qseB , Δ qseBC and qseC Δpr mutants harbouring pDJR29. (a) Graphic representation of the transcriptional reporter plasmids pDJR29. (b)

Techniques Used: Activity Assay

11) Product Images from "Activation of p53 by scaffold-stabilised expression of Mdm2-binding peptides: visualisation of reporter gene induction at the single-cell level"

Article Title: Activation of p53 by scaffold-stabilised expression of Mdm2-binding peptides: visualisation of reporter gene induction at the single-cell level

Journal: British Journal of Cancer

doi: 10.1038/sj.bjc.6602143

Visualisation of p53 activity at the single-cell level. T22 cells were analysed 68 h after transduction by staining for ΔNGFR ( y -axis) and by assaying for  β -gal activity ( x -axis). Density plots from representative experiments for each vector are shown: ( A ) ncmΔNGFR-CI2-12.1-Ala; ( B ) ncmΔNGFR-CI2-12.1; ( C ) ncmΔNGFR-NLS-CI2-12.1-Ala; ( D ) ncmΔNGFR-NLS-CI2-12.1; ( E ) ncmΔNGFR-huArf and ( F ) and ncmΔNGFR-CI2.
Figure Legend Snippet: Visualisation of p53 activity at the single-cell level. T22 cells were analysed 68 h after transduction by staining for ΔNGFR ( y -axis) and by assaying for β -gal activity ( x -axis). Density plots from representative experiments for each vector are shown: ( A ) ncmΔNGFR-CI2-12.1-Ala; ( B ) ncmΔNGFR-CI2-12.1; ( C ) ncmΔNGFR-NLS-CI2-12.1-Ala; ( D ) ncmΔNGFR-NLS-CI2-12.1; ( E ) ncmΔNGFR-huArf and ( F ) and ncmΔNGFR-CI2.

Techniques Used: Activity Assay, Transduction, Staining, Plasmid Preparation

The 12.1 and the Arf37 peptides are stabilised by the CI2 scaffold. T22 reporter cells were transduced with a panel of retroviral vectors expressing the 12.1 peptide and appropriate controls: ( A ) ncmΔNGFR, ncmΔNGFR-12.1, ncmΔNGFR-12.1-Ala, ncmΔNGFR-CI2-12.1, ncmΔNGFR-CI2-12.1-Ala, ncmΔNGFR-NLS-CI2-12.1 and ncmΔNGFR-NLS-CI2-12.1-Ala, ( B ) ncmΔNGFR-Arf37, ncmΔNGFR-CI2, ncmΔNGFR-CI2-Arf37, ncmΔNGFR-NLS-CI2, ncmΔNGFR-NLS-CI2-Arf37, ncmΔNGFR-NLS-CI2-Arf15 and ncmΔNGFR-huArf. Samples were analysed 68 h after transduction by staining the cells for ΔNGFR and assaying for  β -gal activity. Data are plotted as the percentage of double-positive cells (ΔNGFR and  β -gal) of the total transduced population (ΔNGFR-positive cells). Results from three independent transductions are shown.
Figure Legend Snippet: The 12.1 and the Arf37 peptides are stabilised by the CI2 scaffold. T22 reporter cells were transduced with a panel of retroviral vectors expressing the 12.1 peptide and appropriate controls: ( A ) ncmΔNGFR, ncmΔNGFR-12.1, ncmΔNGFR-12.1-Ala, ncmΔNGFR-CI2-12.1, ncmΔNGFR-CI2-12.1-Ala, ncmΔNGFR-NLS-CI2-12.1 and ncmΔNGFR-NLS-CI2-12.1-Ala, ( B ) ncmΔNGFR-Arf37, ncmΔNGFR-CI2, ncmΔNGFR-CI2-Arf37, ncmΔNGFR-NLS-CI2, ncmΔNGFR-NLS-CI2-Arf37, ncmΔNGFR-NLS-CI2-Arf15 and ncmΔNGFR-huArf. Samples were analysed 68 h after transduction by staining the cells for ΔNGFR and assaying for β -gal activity. Data are plotted as the percentage of double-positive cells (ΔNGFR and β -gal) of the total transduced population (ΔNGFR-positive cells). Results from three independent transductions are shown.

Techniques Used: Transduction, Expressing, Staining, Activity Assay

12) Product Images from "Degradation of Proteins and Starch by Combined Immobilization of Protease, α-Amylase and β-Galactosidase on a Single Electrospun Nanofibrous Membrane"

Article Title: Degradation of Proteins and Starch by Combined Immobilization of Protease, α-Amylase and β-Galactosidase on a Single Electrospun Nanofibrous Membrane

Journal: Molecules

doi: 10.3390/molecules24030508

Activity progress curves for β-gal immobilized individually on an SMA nanofibrous mat, and in combination with α-amylase, as well as a combination of α-amylase and protease. Activity was spectrophotometrically determined as a function of time using ONPG as substrate. All results are shown as the mean ± SD of triplicate experiments (n = 3).
Figure Legend Snippet: Activity progress curves for β-gal immobilized individually on an SMA nanofibrous mat, and in combination with α-amylase, as well as a combination of α-amylase and protease. Activity was spectrophotometrically determined as a function of time using ONPG as substrate. All results are shown as the mean ± SD of triplicate experiments (n = 3).

Techniques Used: Activity Assay

13) Product Images from "Diamagnetic Imaging Agents with a Modular Chemical Design for Quantitative Detection of β-Galactosidase and β-Glucuronidase Activities with CatalyCEST MRI"

Article Title: Diamagnetic Imaging Agents with a Modular Chemical Design for Quantitative Detection of β-Galactosidase and β-Glucuronidase Activities with CatalyCEST MRI

Journal: Bioconjugate chemistry

doi: 10.1021/acs.bioconjchem.6b00482

catalyCEST MRI. (a) The experimental CEST spectra (blue circles), the Lorentzian line fitting of the experimental CEST spectra (blue lines), and the Lorentzian line shapes showed two CEST signals from the substrate 6a (solid red lines) and only one CEST signal for the product after β -gal catalysis (dashed red line). (b) The normalized CEST signal at 4.25 ppm decreased after treatment of 6a with β -gal (red). No change in CEST signal was observed after treatment with β -gus (green), with β -gal inhibited by PETG (purple), or in the absence of enzyme (blue). (c,d) Similar results were obtained before and after enzyme catalysis of 6b with β -gus.
Figure Legend Snippet: catalyCEST MRI. (a) The experimental CEST spectra (blue circles), the Lorentzian line fitting of the experimental CEST spectra (blue lines), and the Lorentzian line shapes showed two CEST signals from the substrate 6a (solid red lines) and only one CEST signal for the product after β -gal catalysis (dashed red line). (b) The normalized CEST signal at 4.25 ppm decreased after treatment of 6a with β -gal (red). No change in CEST signal was observed after treatment with β -gus (green), with β -gal inhibited by PETG (purple), or in the absence of enzyme (blue). (c,d) Similar results were obtained before and after enzyme catalysis of 6b with β -gus.

Techniques Used: Magnetic Resonance Imaging

Michaelis–Menten kinetics studies with absorbance at 425 nm. (a) The absorbance at 425 nm was correlated with the concentration of 4-hydroxy-3-nitrobenzyl alcohol ( 1 ) using the Beer–Lambert law. (b) The initial velocity, v i , was determined by monitoring the change in UV absorbance of 6a after the addition of β -gal enzyme and converting the absorbance at 425 nm to concentration using the calibration in panel a. (c) A Hanes–Woolf plot with initial velocities and substrate concentrations was used to determine Michaelis–Menten kinetics parameters. (d,e) This analysis was repeated for evaluating the kinetics of β -gus with its substrate 6b .
Figure Legend Snippet: Michaelis–Menten kinetics studies with absorbance at 425 nm. (a) The absorbance at 425 nm was correlated with the concentration of 4-hydroxy-3-nitrobenzyl alcohol ( 1 ) using the Beer–Lambert law. (b) The initial velocity, v i , was determined by monitoring the change in UV absorbance of 6a after the addition of β -gal enzyme and converting the absorbance at 425 nm to concentration using the calibration in panel a. (c) A Hanes–Woolf plot with initial velocities and substrate concentrations was used to determine Michaelis–Menten kinetics parameters. (d,e) This analysis was repeated for evaluating the kinetics of β -gus with its substrate 6b .

Techniques Used: Concentration Assay

14) Product Images from "Adenoviral Vector-Mediated Expression of B-50/GAP-43 Induces Alterations in the Membrane Organization of Olfactory Axon TerminalsIn Vivo"

Article Title: Adenoviral Vector-Mediated Expression of B-50/GAP-43 Induces Alterations in the Membrane Organization of Olfactory Axon TerminalsIn Vivo

Journal: The Journal of Neuroscience

doi: 10.1523/JNEUROSCI.17-17-06575.1997

Ad- B-50/GAP-43 - and Ad- LacZ -directed expression of B-50/GAP-43 and β-gal in mature olfactory neurons. A, B , Low power photomicrographs of transversal sections of mouse olfactory epithelia stained for β-gal ( A ) and B-50/GAP-43 ( B ) showing
Figure Legend Snippet: Ad- B-50/GAP-43 - and Ad- LacZ -directed expression of B-50/GAP-43 and β-gal in mature olfactory neurons. A, B , Low power photomicrographs of transversal sections of mouse olfactory epithelia stained for β-gal ( A ) and B-50/GAP-43 ( B ) showing

Techniques Used: Expressing, Staining

15) Product Images from "The matricellular protein TSP1 promotes human and mouse endothelial cell senescence through CD47 and Nox1"

Article Title: The matricellular protein TSP1 promotes human and mouse endothelial cell senescence through CD47 and Nox1

Journal: Science signaling

doi: 10.1126/scisignal.aaj1784

Potential clinical relevance of Nox1-mediated endothelial senescence in human pulmonary vascular tissue Data points for human tissue are shown in green; data for cell experiments are shown in red; data in gray are the corresponding controls. Data for ( A ) to ( E ) are plotted as linear regression ( n = 8 samples); equation, r 2 , and P values are indicated in the corresponding graph. (A and B) Correlation between NADPH-driven O 2 •− production, as measured by cytochrome c reduction (A), or H 2 O 2 production, as measured by Amplex Red fluorescence (B), and age in human lung homogenates. (C and D) Abundance of Nox1 (C) and p53 and p21 cip (D), as measured by Western blot of total homogenates of aging human lung. ( E ) Intimal immunofluorescence for Nox1 (top, green) and p21 cip (bottom, red) in aged human lung sections. Scale bar, 50 μm. Graphs show linear regression analyses. ( F ) HPAEC senescence induced by TSP1 in the presence or absence of NoxA1ds, as measured by SA-β-Gal staining. Graphical data are means ± SEM ( n = 3 biological replicates per treatment). Scale bar, 40 μm. * P
Figure Legend Snippet: Potential clinical relevance of Nox1-mediated endothelial senescence in human pulmonary vascular tissue Data points for human tissue are shown in green; data for cell experiments are shown in red; data in gray are the corresponding controls. Data for ( A ) to ( E ) are plotted as linear regression ( n = 8 samples); equation, r 2 , and P values are indicated in the corresponding graph. (A and B) Correlation between NADPH-driven O 2 •− production, as measured by cytochrome c reduction (A), or H 2 O 2 production, as measured by Amplex Red fluorescence (B), and age in human lung homogenates. (C and D) Abundance of Nox1 (C) and p53 and p21 cip (D), as measured by Western blot of total homogenates of aging human lung. ( E ) Intimal immunofluorescence for Nox1 (top, green) and p21 cip (bottom, red) in aged human lung sections. Scale bar, 50 μm. Graphs show linear regression analyses. ( F ) HPAEC senescence induced by TSP1 in the presence or absence of NoxA1ds, as measured by SA-β-Gal staining. Graphical data are means ± SEM ( n = 3 biological replicates per treatment). Scale bar, 40 μm. * P

Techniques Used: Fluorescence, Western Blot, Immunofluorescence, Staining

Nox1 as a potential therapeutic target to inhibit matricellular-mediated endothelial senescence Data for Nox1 OE are shown in blue; data for Nox1 − / − samples are shown in orange; data for cell experiments are shown in red; data in gray are the corresponding controls. ( A and B ) Effect of human Nox1 OE on SA-β-Gal staining and O 2 •− production. Graphical data are means ± SEM ( n = 3 biological replicates per treatment). Scale bar, 40 μm. * P
Figure Legend Snippet: Nox1 as a potential therapeutic target to inhibit matricellular-mediated endothelial senescence Data for Nox1 OE are shown in blue; data for Nox1 − / − samples are shown in orange; data for cell experiments are shown in red; data in gray are the corresponding controls. ( A and B ) Effect of human Nox1 OE on SA-β-Gal staining and O 2 •− production. Graphical data are means ± SEM ( n = 3 biological replicates per treatment). Scale bar, 40 μm. * P

Techniques Used: Staining

16) Product Images from "A Mechanistic Proof-of-concept Clinical Trial With JX-594, a Targeted Multi-mechanistic Oncolytic Poxvirus, in Patients With Metastatic Melanoma"

Article Title: A Mechanistic Proof-of-concept Clinical Trial With JX-594, a Targeted Multi-mechanistic Oncolytic Poxvirus, in Patients With Metastatic Melanoma

Journal: Molecular Therapy

doi: 10.1038/mt.2011.132

Antibody induction to JX-594 and to the β-gal marker transgene product . ( a ) Neutralizing antibody titers over time following treatment initiation; individual data points are circles and the line is the mean. ( b ) Anti-β-gal antibody titers
Figure Legend Snippet: Antibody induction to JX-594 and to the β-gal marker transgene product . ( a ) Neutralizing antibody titers over time following treatment initiation; individual data points are circles and the line is the mean. ( b ) Anti-β-gal antibody titers

Techniques Used: Marker

17) Product Images from "Promotion of Dendritic Growth by CPG15, an Activity-Induced Signaling Molecule"

Article Title: Promotion of Dendritic Growth by CPG15, an Activity-Induced Signaling Molecule

Journal: Science (New York, N.Y.)

doi:

Quantification of CPG15 growth-promoting effect on dendritic arbors. ( A ) The average TDBL of rostrally projecting CPG15 neurons, β-gal neurons, and uninfected control neurons is plotted over 3 days of imaging. On the first day of imaging, the average TDBL of projection neurons from CPG15VV-infected animals was 447 ± 69 μm ( n = 39), significantly larger ( P
Figure Legend Snippet: Quantification of CPG15 growth-promoting effect on dendritic arbors. ( A ) The average TDBL of rostrally projecting CPG15 neurons, β-gal neurons, and uninfected control neurons is plotted over 3 days of imaging. On the first day of imaging, the average TDBL of projection neurons from CPG15VV-infected animals was 447 ± 69 μm ( n = 39), significantly larger ( P

Techniques Used: Imaging, Infection

CPG15 induction by kainic acid and its expression in Xenopus optic tectum. ( A ) Immuno-blot of protein extracts from tadpoles harvested at the specified times after intraventricular injection of KA, or rat hippocampus dentate gyri 24 hours after ip injection of KA (right lane). Incubation with the antiserum to CPG15 labels a 12-kD band (arrow) that is not seen with preimmune serum (PI). Confocal images of sections through the optic tecti of untreated tadpoles ( B and C ) or tadpoles infected with CPG15VV ( D and E ) or CPG15t3VV ( F and G ). Sections probed with preimmune rabbit serum show no specific labeling (B). Outlined on this section are the optic tectal neuropil (N), differentiated tectal neurons (TN), and the proliferative zone (PZ). These same regions can be discerned in the sections stained with the antisera to CPG15 [(C), (E), and (G)]. A honeycomb pattern of endogenous CPG15 immunoreactivity can be seen in the TN region of the tectum, and retinal ganglion cell axons are stained in N (C). Sections from animals infected with virus were double-labeled with anti– β-gal to show extent of infection [(D) and (F)] and with anti-CPG15 at higher magnification [(E) and (G)]. In the infected tecti [(E) and (G)], the honeycomb pattern of CPG15 immunoreactivity also extends into the PZ, where many infected neurons are located [(D) and (F)]. Arrows mark retinotectal axons. Bar, 100 μm for upper panel and 50 μm for lower panels.
Figure Legend Snippet: CPG15 induction by kainic acid and its expression in Xenopus optic tectum. ( A ) Immuno-blot of protein extracts from tadpoles harvested at the specified times after intraventricular injection of KA, or rat hippocampus dentate gyri 24 hours after ip injection of KA (right lane). Incubation with the antiserum to CPG15 labels a 12-kD band (arrow) that is not seen with preimmune serum (PI). Confocal images of sections through the optic tecti of untreated tadpoles ( B and C ) or tadpoles infected with CPG15VV ( D and E ) or CPG15t3VV ( F and G ). Sections probed with preimmune rabbit serum show no specific labeling (B). Outlined on this section are the optic tectal neuropil (N), differentiated tectal neurons (TN), and the proliferative zone (PZ). These same regions can be discerned in the sections stained with the antisera to CPG15 [(C), (E), and (G)]. A honeycomb pattern of endogenous CPG15 immunoreactivity can be seen in the TN region of the tectum, and retinal ganglion cell axons are stained in N (C). Sections from animals infected with virus were double-labeled with anti– β-gal to show extent of infection [(D) and (F)] and with anti-CPG15 at higher magnification [(E) and (G)]. In the infected tecti [(E) and (G)], the honeycomb pattern of CPG15 immunoreactivity also extends into the PZ, where many infected neurons are located [(D) and (F)]. Arrows mark retinotectal axons. Bar, 100 μm for upper panel and 50 μm for lower panels.

Techniques Used: Expressing, Injection, Incubation, Infection, Labeling, Staining

CPG15 does not affect tectal interneurons. ( A ) Drawings of interneurons from β-gal– infected animals (left) and CPG15VV-infected animals (right) with a TDBL closest to the mean branch length of each group. ( B ) The growth rate (TDBL on day 2 – TDBL on day 1) is significantly greater ( P
Figure Legend Snippet: CPG15 does not affect tectal interneurons. ( A ) Drawings of interneurons from β-gal– infected animals (left) and CPG15VV-infected animals (right) with a TDBL closest to the mean branch length of each group. ( B ) The growth rate (TDBL on day 2 – TDBL on day 1) is significantly greater ( P

Techniques Used: Infection

CPG15 promotes growth through intercellular signaling. ( A ) A 3D reconstruction of a tectal projection neuron from a CPG15VV-infected animal, with a TDBL of 1684 μm on the first day of imaging (day 1) and 2021 μm on the second day (day 2). This cell is a clear outlier on both days as the largest control cell is 642 μm on the first day of imaging and 1017 μm on the second. ( B ) Top panel shows a drawing of the tadpole optic tectum (OT) and the tectal ventricle (V) with the marked location of this cell. The green square delineates the region shown in the bottom panel. Bottom panel shows a superimposition of images collected with a 488-nm filter visualizing the DiI-labeled cell imaged in green and images collected with a 647-nm filter visualizing β-gal immunopositive cells in red. The arrow marks the cell imaged in (A).
Figure Legend Snippet: CPG15 promotes growth through intercellular signaling. ( A ) A 3D reconstruction of a tectal projection neuron from a CPG15VV-infected animal, with a TDBL of 1684 μm on the first day of imaging (day 1) and 2021 μm on the second day (day 2). This cell is a clear outlier on both days as the largest control cell is 642 μm on the first day of imaging and 1017 μm on the second. ( B ) Top panel shows a drawing of the tadpole optic tectum (OT) and the tectal ventricle (V) with the marked location of this cell. The green square delineates the region shown in the bottom panel. Bottom panel shows a superimposition of images collected with a 488-nm filter visualizing the DiI-labeled cell imaged in green and images collected with a 647-nm filter visualizing β-gal immunopositive cells in red. The arrow marks the cell imaged in (A).

Techniques Used: Infection, Imaging, Labeling

18) Product Images from "Efficient Nonviral Gene Therapy Using Folate-Targeted Chitosan-DNA Nanoparticles In Vitro"

Article Title: Efficient Nonviral Gene Therapy Using Folate-Targeted Chitosan-DNA Nanoparticles In Vitro

Journal: ISRN Pharmaceutics

doi: 10.5402/2012/369270

Cells viability treated with chitosan nanoparticules (a) containing the VR1412  β -gal plasmid DNA. Group 1: negative control (KB cells); Group 2: naked DNA; Group 3: Ch-DNA nanoparticule with a Mw = 5 KDa chitosan; Group 4: Ch-PEG-FA-DNA nanoparticule with a Mw = 5 KDa chitosan; Group 5: ch-DNA nanoparticule with a Mw = 25 KDa chitosan; Group 6: Ch-PEG-FA-DNA with a Mw = 25 KDa chitosan; Group 7: Ch-DNA with a Mw = 50 KDa chitosan; Group 8: Ch-PEG-FA-DNA nanoparticule with a Mw = 50 KDa chitosan; Group 9: Lipofectamine. (b) Chitosan nonconjugated with DNA, Group 1: negative control; Group 3: Mw = 5 KDa chitosan; Group 4: Mw = 5 KDa chitosan combined with FA; Group 5: Mw = 25 KDa Chitosan; Group 6: Mw = 25 KDa chitosan combined with FA; Group 7: Mw = 50 KDa chitosan; Group 8: Mw = 50 KDa chitosan combined with FA; Group 9: lipofectamine. *Statistical significant differences compared with positive control ( P
Figure Legend Snippet: Cells viability treated with chitosan nanoparticules (a) containing the VR1412 β -gal plasmid DNA. Group 1: negative control (KB cells); Group 2: naked DNA; Group 3: Ch-DNA nanoparticule with a Mw = 5 KDa chitosan; Group 4: Ch-PEG-FA-DNA nanoparticule with a Mw = 5 KDa chitosan; Group 5: ch-DNA nanoparticule with a Mw = 25 KDa chitosan; Group 6: Ch-PEG-FA-DNA with a Mw = 25 KDa chitosan; Group 7: Ch-DNA with a Mw = 50 KDa chitosan; Group 8: Ch-PEG-FA-DNA nanoparticule with a Mw = 50 KDa chitosan; Group 9: Lipofectamine. (b) Chitosan nonconjugated with DNA, Group 1: negative control; Group 3: Mw = 5 KDa chitosan; Group 4: Mw = 5 KDa chitosan combined with FA; Group 5: Mw = 25 KDa Chitosan; Group 6: Mw = 25 KDa chitosan combined with FA; Group 7: Mw = 50 KDa chitosan; Group 8: Mw = 50 KDa chitosan combined with FA; Group 9: lipofectamine. *Statistical significant differences compared with positive control ( P

Techniques Used: Plasmid Preparation, Negative Control, Positive Control

Expression of (a) VR1412 β -gal, (b) GFP in KB cells and (c) KB cells expressing GFP seen with fluorescent microscopy (magnification × 10) Group 1: negative control (KB cells); Group 2: Naked DNA; Group 3: Ch-DNA nanoparticule with a Mw = 5 KDa chitosan; Group 4: Ch-PEG-FA-DNA nanoparticule with a Mw = 5 KDa chitosan; Group 5: Ch-DNA nanoparticule with a Mw = 25 KDa chitosan; Group 6 : Ch-PEG-FA-DNA with a Mw = 25 KDa chitosan; Group 7: Ch-DNA with a Mw = 50 KDa chitosan; Group 8: Ch-PEG-FA-DNA nanoparticulewith a Mw = 50 KDa chitosan; Group 9: lipofectamine coupled with DNA; Group 10: KB cells seen with optical microscopy; Group 11: KB cells seen with fluorescent microscopy; Group 12: negative control, nontreated KB cells seen with fluorescent microscopy. *Statistical significant differences compared with positive control ( P
Figure Legend Snippet: Expression of (a) VR1412 β -gal, (b) GFP in KB cells and (c) KB cells expressing GFP seen with fluorescent microscopy (magnification × 10) Group 1: negative control (KB cells); Group 2: Naked DNA; Group 3: Ch-DNA nanoparticule with a Mw = 5 KDa chitosan; Group 4: Ch-PEG-FA-DNA nanoparticule with a Mw = 5 KDa chitosan; Group 5: Ch-DNA nanoparticule with a Mw = 25 KDa chitosan; Group 6 : Ch-PEG-FA-DNA with a Mw = 25 KDa chitosan; Group 7: Ch-DNA with a Mw = 50 KDa chitosan; Group 8: Ch-PEG-FA-DNA nanoparticulewith a Mw = 50 KDa chitosan; Group 9: lipofectamine coupled with DNA; Group 10: KB cells seen with optical microscopy; Group 11: KB cells seen with fluorescent microscopy; Group 12: negative control, nontreated KB cells seen with fluorescent microscopy. *Statistical significant differences compared with positive control ( P

Techniques Used: Expressing, Microscopy, Negative Control, Positive Control

Agarose gel electrophoresis. (a) Chitosan nanoparticles with VR1412  β -gal plasmid DNA. (b) Chitosan nanoparticles with GFP plasmid DNA. Lane 1: ladder; lane 2: naked DNA; lane 3: Ch-DNA nanoparticule with a Mw = 5 KDa chitosan; lane 4: Ch-PEG-FA-DNA nanoparticule with a Mw = 5 KDa chitosan; lane 5: Ch-DNA nanoparticule with a Mw = 25 KDa chitosan; lane 6: Ch-PEG-FA-DNA with a Mw = 25 KDa chitosan; lane 7: Ch-DNA with a Mw = 50 KDa chitosan; lane 8: Ch-PEG-FA-DNA nanoparticule with a Mw = 50 KDa chitosan.
Figure Legend Snippet: Agarose gel electrophoresis. (a) Chitosan nanoparticles with VR1412 β -gal plasmid DNA. (b) Chitosan nanoparticles with GFP plasmid DNA. Lane 1: ladder; lane 2: naked DNA; lane 3: Ch-DNA nanoparticule with a Mw = 5 KDa chitosan; lane 4: Ch-PEG-FA-DNA nanoparticule with a Mw = 5 KDa chitosan; lane 5: Ch-DNA nanoparticule with a Mw = 25 KDa chitosan; lane 6: Ch-PEG-FA-DNA with a Mw = 25 KDa chitosan; lane 7: Ch-DNA with a Mw = 50 KDa chitosan; lane 8: Ch-PEG-FA-DNA nanoparticule with a Mw = 50 KDa chitosan.

Techniques Used: Agarose Gel Electrophoresis, Plasmid Preparation

Distribution function of nanoparticles sizes (diameter in nm). (1) Ch5- β -gal: 114.27 ± 20.08 nm; (2) Ch5-PEG-FA- β -gal: 150.03 ± 9.76 nm; (3) Ch5-GFP: 223.83 ± 11.54 nm; (4) Ch5-PEG-FA-GFP: 278 ± 27.22 nm; (5) Ch25- β -gal: 127.67 ± 16.21 nm; (6) Ch25-PEG-FA- β -gal: 134.84 ± 14.13 nm; (7) Ch25-GFP: 151.43 ± 9.35 nm; (8) Ch25-PEG-FA-GFP: 204.33 ± 5.91 nm; (9) Ch50- β -gal: 111.94 ± 20.75 nm; (10) Ch50-PEG-FA- β -gal: 247.34 ± 18.33 nm; (11) Ch50-GFP: 160.21 ± 6.82 nm; (12) Ch50-PEG-FA-GFP: 302.08 ± 34.13 nm ( P
Figure Legend Snippet: Distribution function of nanoparticles sizes (diameter in nm). (1) Ch5- β -gal: 114.27 ± 20.08 nm; (2) Ch5-PEG-FA- β -gal: 150.03 ± 9.76 nm; (3) Ch5-GFP: 223.83 ± 11.54 nm; (4) Ch5-PEG-FA-GFP: 278 ± 27.22 nm; (5) Ch25- β -gal: 127.67 ± 16.21 nm; (6) Ch25-PEG-FA- β -gal: 134.84 ± 14.13 nm; (7) Ch25-GFP: 151.43 ± 9.35 nm; (8) Ch25-PEG-FA-GFP: 204.33 ± 5.91 nm; (9) Ch50- β -gal: 111.94 ± 20.75 nm; (10) Ch50-PEG-FA- β -gal: 247.34 ± 18.33 nm; (11) Ch50-GFP: 160.21 ± 6.82 nm; (12) Ch50-PEG-FA-GFP: 302.08 ± 34.13 nm ( P

Techniques Used:

19) Product Images from "The mouse C9ORF72 ortholog is enriched in neurons known to degenerate in ALS and FTD"

Article Title: The mouse C9ORF72 ortholog is enriched in neurons known to degenerate in ALS and FTD

Journal: Nature neuroscience

doi: 10.1038/nn.3566

Characterization of the cells expressing β-gal under control of the C9ORF72 -ortholog promoter (a). Co-localization of β-gal, CTIP2 (layer V) and NeuN in cortex. Green-β-gal, red-CTIP2, blue-NeuN. (b) shows Z-stack and orthogonal images of white rectangle in (a). (c) Co-localization of β-gal with ChAT, and β-gal with NeuN in ventral horn of the spinal cord are shown. Dashed line represents the border between grey and white matter. (d) β-gal positive cells are not colocalized with GFAP or Iba1 in ventral horn of the spinal cord. (e-j) Spliced C9ORF72 mRNA is specifically expressed in motor neuron-like cells in the mouse (e) and human spinal cord (f-j). (f) Composite photomicrograph of a coronal section of a human spinal cord stained with Hoechst to visualize DNA. (g) Map of C9ORF72 -expressing cells (black dots) observed in the spinal cord section show in panels (f-j). Labeled cells were sparse and were confined to cell bodies distributed in the ventral and lateral horns of the spinal cord. The wedge-shaped tear on the left dorsolateral edge of the spinal cord is a histological artifact. (h) Photomicrograph of the yellow-boxed region in f, showing the in situ hybridization signal of DIG-labeled antisense riboprobe against spliced human C9ORF72 mRNA (yellow arrows). Hybridization was strong and specific. (i) High power photomicrograph of the green boxed region in (h), illustrating a representative labeled cell (green), whose nucleus (arrowhead) is considerably larger than those of surrounding cells (yellow arrows). (i and j) Representative C9ORF72 -expressing cells are large and pyramidal, and strongly reminiscent of spinal cord motor neurons. Bar, 50 μm (a, e) and 20 μm (b, c, d, h), 2mm (f), 100μm (i), and 10μm (j).
Figure Legend Snippet: Characterization of the cells expressing β-gal under control of the C9ORF72 -ortholog promoter (a). Co-localization of β-gal, CTIP2 (layer V) and NeuN in cortex. Green-β-gal, red-CTIP2, blue-NeuN. (b) shows Z-stack and orthogonal images of white rectangle in (a). (c) Co-localization of β-gal with ChAT, and β-gal with NeuN in ventral horn of the spinal cord are shown. Dashed line represents the border between grey and white matter. (d) β-gal positive cells are not colocalized with GFAP or Iba1 in ventral horn of the spinal cord. (e-j) Spliced C9ORF72 mRNA is specifically expressed in motor neuron-like cells in the mouse (e) and human spinal cord (f-j). (f) Composite photomicrograph of a coronal section of a human spinal cord stained with Hoechst to visualize DNA. (g) Map of C9ORF72 -expressing cells (black dots) observed in the spinal cord section show in panels (f-j). Labeled cells were sparse and were confined to cell bodies distributed in the ventral and lateral horns of the spinal cord. The wedge-shaped tear on the left dorsolateral edge of the spinal cord is a histological artifact. (h) Photomicrograph of the yellow-boxed region in f, showing the in situ hybridization signal of DIG-labeled antisense riboprobe against spliced human C9ORF72 mRNA (yellow arrows). Hybridization was strong and specific. (i) High power photomicrograph of the green boxed region in (h), illustrating a representative labeled cell (green), whose nucleus (arrowhead) is considerably larger than those of surrounding cells (yellow arrows). (i and j) Representative C9ORF72 -expressing cells are large and pyramidal, and strongly reminiscent of spinal cord motor neurons. Bar, 50 μm (a, e) and 20 μm (b, c, d, h), 2mm (f), 100μm (i), and 10μm (j).

Techniques Used: Expressing, Staining, Labeling, In Situ Hybridization, Hybridization

20) Product Images from "Nose-to-brain delivery of macromolecules mediated by cell-penetrating peptides"

Article Title: Nose-to-brain delivery of macromolecules mediated by cell-penetrating peptides

Journal: Acta Pharmaceutica Sinica. B

doi: 10.1016/j.apsb.2016.04.001

Enzymatic activity of HPR (left panel) and β -gal (right panel). LMWP-linked proteins were represented by grey bars, and native proteins white bars. LMWP-enzymes were significantly higher than their non-modified counterparts ( n =3).
Figure Legend Snippet: Enzymatic activity of HPR (left panel) and β -gal (right panel). LMWP-linked proteins were represented by grey bars, and native proteins white bars. LMWP-enzymes were significantly higher than their non-modified counterparts ( n =3).

Techniques Used: Activity Assay, Modification

21) Product Images from "Accelerated Hippocampal Spreading Depression and Enhanced Locomotory Activity in Mice with Astrocyte-Directed Inactivation of Connexin43"

Article Title: Accelerated Hippocampal Spreading Depression and Enhanced Locomotory Activity in Mice with Astrocyte-Directed Inactivation of Connexin43

Journal: The Journal of Neuroscience

doi: 10.1523/JNEUROSCI.23-03-00766.2003

Concept of lacZ activation on Cx43 deletion. Cre-mediated deletion of floxed DNA at the Cx43 locus leads to lacZ expression in cells that show Cx43 gene activity. A , Cx43 genomic DNA and mRNA and protein expression of the Cx43 fl allele. Top row , Genomic DNA. Thin line , genomic DNA; black boxes , noncoding part of Cx43 exon 2; dark gray box , Cx43 -coding region ( Cx43 cdr ); white box , selection marker DNA in reverse orientation to Cx43 and lacZ ; light gray box , lacZ reporter gene with nuclear localization signal ( NLS ) and polyadenylation signal ( SV40 pA ); triangles , loxP sites; bar , 1 kb; H , Hin dIII; cdr , coding region; tk , Herpes simplex virus thymidine-kinase; neo , neomycin-phosphotransferase. Bottom row , mRNA processing. Horizontal line , Exonic RNA; inclined line , intronic RNA; AAAA , polyadenylated mRNA tail; ellipses , ribosomes. Horizontal arrow delineates translated mRNA. Small vertical arrow delineates translation process. Large vertical arrow delineates cre-mediated deletion. B , Cx43 genomic DNA and mRNA and protein expression of the Cx43 del allele. For explanations, see A . Circles indicate β-galactosidase peptides. C , X-Gal-stained 13.5 dpc embryos; LacZ expression in the developing brain of a Cx43 fl/+ , hGFAP-cre embryo and nonstained Cx43 fl/+ control embryo.
Figure Legend Snippet: Concept of lacZ activation on Cx43 deletion. Cre-mediated deletion of floxed DNA at the Cx43 locus leads to lacZ expression in cells that show Cx43 gene activity. A , Cx43 genomic DNA and mRNA and protein expression of the Cx43 fl allele. Top row , Genomic DNA. Thin line , genomic DNA; black boxes , noncoding part of Cx43 exon 2; dark gray box , Cx43 -coding region ( Cx43 cdr ); white box , selection marker DNA in reverse orientation to Cx43 and lacZ ; light gray box , lacZ reporter gene with nuclear localization signal ( NLS ) and polyadenylation signal ( SV40 pA ); triangles , loxP sites; bar , 1 kb; H , Hin dIII; cdr , coding region; tk , Herpes simplex virus thymidine-kinase; neo , neomycin-phosphotransferase. Bottom row , mRNA processing. Horizontal line , Exonic RNA; inclined line , intronic RNA; AAAA , polyadenylated mRNA tail; ellipses , ribosomes. Horizontal arrow delineates translated mRNA. Small vertical arrow delineates translation process. Large vertical arrow delineates cre-mediated deletion. B , Cx43 genomic DNA and mRNA and protein expression of the Cx43 del allele. For explanations, see A . Circles indicate β-galactosidase peptides. C , X-Gal-stained 13.5 dpc embryos; LacZ expression in the developing brain of a Cx43 fl/+ , hGFAP-cre embryo and nonstained Cx43 fl/+ control embryo.

Techniques Used: Activation Assay, Expressing, Activity Assay, Selection, Marker, Staining

Cell-type specificity of hGFAP-cre -mediated Cx43 inactivation monitored by lacZ expression. A – D , Double-immunofluorescence analysis on hippocampal cryosections of Cx43 fl/+ , hGFAP-cre mice ( A, B ) and Cx43 del/+ mice ( C, D ) using antibodies directed to β-galactosidase ( red ) and GFAP ( A , C , green ) or NeuN ( B , D , green ). Coexpression of β-galactosidase with GFAP ( A ) but not with NeuN ( B ) occurs in Cx43 fl/+ , hGFAP-cre mice indistinguishable from that in Cx43 del/+ mice ( C and D , respectively). Scale bar (shown in D ): 50 μm. E – P , X-Gal staining of brain cryosections from mice with general ( E , I , M , Cx43 del/+ ), astrocyte-specific ( F , J , N , H , Cx43 fl/- , hGFAP-cre), endothelial cell-specific deletion of the Cx43 fl allele ( G , K , O , L , Cx43 fl/- , TIE2-cre), and multiple cre-transgenic mice ( P , Cx43 fl/- , hGFAP-cre, TIE2-cre). Scale bar (shown in P ): E – O , 140 μm; P , 24 μm. E – G , Cortex. I – K , Hippocampus. White dots demarcate granule cells of the dentate gyrus and pyramidal cells. H , L , Ventricles. Arrow indicates choroid plexus. M – P , Cerebellum. Asterisk indicates molecular layer. P , Purkinje cell layer; G , granule cell layer; M , white matter. Arrows in M and N indicate leptomeningeal cell layer. Arrow in O indicates streak-like arrangement of lacZ -expressing cells indicative of blood vessels. Arrows in P indicate nonexpressing Purkinje cell bodies.
Figure Legend Snippet: Cell-type specificity of hGFAP-cre -mediated Cx43 inactivation monitored by lacZ expression. A – D , Double-immunofluorescence analysis on hippocampal cryosections of Cx43 fl/+ , hGFAP-cre mice ( A, B ) and Cx43 del/+ mice ( C, D ) using antibodies directed to β-galactosidase ( red ) and GFAP ( A , C , green ) or NeuN ( B , D , green ). Coexpression of β-galactosidase with GFAP ( A ) but not with NeuN ( B ) occurs in Cx43 fl/+ , hGFAP-cre mice indistinguishable from that in Cx43 del/+ mice ( C and D , respectively). Scale bar (shown in D ): 50 μm. E – P , X-Gal staining of brain cryosections from mice with general ( E , I , M , Cx43 del/+ ), astrocyte-specific ( F , J , N , H , Cx43 fl/- , hGFAP-cre), endothelial cell-specific deletion of the Cx43 fl allele ( G , K , O , L , Cx43 fl/- , TIE2-cre), and multiple cre-transgenic mice ( P , Cx43 fl/- , hGFAP-cre, TIE2-cre). Scale bar (shown in P ): E – O , 140 μm; P , 24 μm. E – G , Cortex. I – K , Hippocampus. White dots demarcate granule cells of the dentate gyrus and pyramidal cells. H , L , Ventricles. Arrow indicates choroid plexus. M – P , Cerebellum. Asterisk indicates molecular layer. P , Purkinje cell layer; G , granule cell layer; M , white matter. Arrows in M and N indicate leptomeningeal cell layer. Arrow in O indicates streak-like arrangement of lacZ -expressing cells indicative of blood vessels. Arrows in P indicate nonexpressing Purkinje cell bodies.

Techniques Used: Expressing, Immunofluorescence, Mouse Assay, Staining, Transgenic Assay

22) Product Images from "Expression, purification, crystallization and preliminary X-ray crystallographic analysis of human ?-galactosidase"

Article Title: Expression, purification, crystallization and preliminary X-ray crystallographic analysis of human ?-galactosidase

Journal: Acta Crystallographica Section F: Structural Biology and Crystallization Communications

doi: 10.1107/S1744309111047920

A β-Gal–galactose crystal in the cryoloop used for data collection.
Figure Legend Snippet: A β-Gal–galactose crystal in the cryoloop used for data collection.

Techniques Used:

SDS–PAGE analysis of β-Gal purification. Lane M , molecular markers (labelled in kDa); lane 1, culture supernatant; lane 2, β-Gal after Phenyl Sepharose purification; lane 3, β-Gal after Q Sepharose purification;
Figure Legend Snippet: SDS–PAGE analysis of β-Gal purification. Lane M , molecular markers (labelled in kDa); lane 1, culture supernatant; lane 2, β-Gal after Phenyl Sepharose purification; lane 3, β-Gal after Q Sepharose purification;

Techniques Used: SDS Page, Purification

23) Product Images from "Feline Immunodeficiency Virus as a Gene Transfer Vector in the Rat Nucleus Tractus Solitarii"

Article Title: Feline Immunodeficiency Virus as a Gene Transfer Vector in the Rat Nucleus Tractus Solitarii

Journal: Cellular and molecular neurobiology

doi: 10.1007/s10571-009-9456-5

Confocal images ( in gray scale ) of the ipsilateral NG with immunofluorescent staining for β Gal and PGP9.5 after FIVLacZ or AdLacZ had been injected unilaterally into the NTS. Neither cells nor fibers labeled for β Gal-IR are seen in the ipsilateral NG after FIVLacZ had been injected to the NTS, as shown in a. c is a confocal image of PGP9.5-IR of the same section to show the structure of the NG section in a. b is a confocal image of NG showing cells (indicated by arrows ) and fibers expressing β Gal-IR after injection of AdLacZ into the NTS. d is confocal image of PGP9.5 showing some NG cells ( arrows ) that are double labeled for β Gal-IR and PGP9.5-IR. Scale bar = 100 μm
Figure Legend Snippet: Confocal images ( in gray scale ) of the ipsilateral NG with immunofluorescent staining for β Gal and PGP9.5 after FIVLacZ or AdLacZ had been injected unilaterally into the NTS. Neither cells nor fibers labeled for β Gal-IR are seen in the ipsilateral NG after FIVLacZ had been injected to the NTS, as shown in a. c is a confocal image of PGP9.5-IR of the same section to show the structure of the NG section in a. b is a confocal image of NG showing cells (indicated by arrows ) and fibers expressing β Gal-IR after injection of AdLacZ into the NTS. d is confocal image of PGP9.5 showing some NG cells ( arrows ) that are double labeled for β Gal-IR and PGP9.5-IR. Scale bar = 100 μm

Techniques Used: Staining, Injection, Labeling, Expressing

Pseudocolored confocal images of rat NTS after FIVLacZ ( a–f ) had been injected into this nucleus, which was then subjected to double-label immunofluorescent staining for β Gal ( green ) and NeuN ( red ), or for β Gal ( green ) and GFAP ( red ), or for β Gal ( green ) and nNOS ( red ). c is a merged confocal image of a (FIVLacZ transfected, green) and b (NeuN-IR, red ). White arrows in a–c indicate representative cells that are stained for both β Gal-IR and NeuN-IR and thus appear yellow in the merged image. Note that the majority of FIVLacZ transfected NTS cells also contained NeuN-IR. f is the merged confocal image of d (FIVLacZ transfected, green ) and e (GFAP-IR, red ). Blue arrows in d and f indicate cells that are positive for β Gal-IR alone. Empty white arrows in e and f indicate cells or processes that are positive for GFAP-IR alone. The majority of FIVLacZ-transfected NTS cells do not contain GFAP-IR. i is a merged confocal image of G (FIVLacZ transfected, green ) and h (nNOS-IR, red ). White arrows in g–i indicate cells that are stained for both β Gal-IR and nNOS-IR. Scale bar = 20 μm
Figure Legend Snippet: Pseudocolored confocal images of rat NTS after FIVLacZ ( a–f ) had been injected into this nucleus, which was then subjected to double-label immunofluorescent staining for β Gal ( green ) and NeuN ( red ), or for β Gal ( green ) and GFAP ( red ), or for β Gal ( green ) and nNOS ( red ). c is a merged confocal image of a (FIVLacZ transfected, green) and b (NeuN-IR, red ). White arrows in a–c indicate representative cells that are stained for both β Gal-IR and NeuN-IR and thus appear yellow in the merged image. Note that the majority of FIVLacZ transfected NTS cells also contained NeuN-IR. f is the merged confocal image of d (FIVLacZ transfected, green ) and e (GFAP-IR, red ). Blue arrows in d and f indicate cells that are positive for β Gal-IR alone. Empty white arrows in e and f indicate cells or processes that are positive for GFAP-IR alone. The majority of FIVLacZ-transfected NTS cells do not contain GFAP-IR. i is a merged confocal image of G (FIVLacZ transfected, green ) and h (nNOS-IR, red ). White arrows in g–i indicate cells that are stained for both β Gal-IR and nNOS-IR. Scale bar = 20 μm

Techniques Used: Injection, Staining, Transfection

Confocal images ( in gray scale ) of the rat NTS after FIVLacZ had been injected unilaterally into this nucleus, which then was subjected to immunofluorescent staining for β Gal ( a, b ) and nNOS ( e, f ). Numerous cells positive for β Gal-IR are observed in the injected side ( b ). A few transfected cells in the adjacent areas, such as the dorsal motor nucleus of vagus (DMV), area postrema (AP), and gracilus nucleus (Gr) were also noted. The contralateral NTS ( a ) shows almost no transfected cells. The schematic sketches of a and b are shown in c and d , respectively. e and f show confocal images of nNOS-IR of the same section as a and b , respectively. Note that the staining pattern and distribution of nNOS-IR in the NTS and in adjacent areas in the injected side ( f ) are similar to that of the contralateral side ( e ). Abbreviation : Tr, tractus solitarius. Scale bar = 100 μm
Figure Legend Snippet: Confocal images ( in gray scale ) of the rat NTS after FIVLacZ had been injected unilaterally into this nucleus, which then was subjected to immunofluorescent staining for β Gal ( a, b ) and nNOS ( e, f ). Numerous cells positive for β Gal-IR are observed in the injected side ( b ). A few transfected cells in the adjacent areas, such as the dorsal motor nucleus of vagus (DMV), area postrema (AP), and gracilus nucleus (Gr) were also noted. The contralateral NTS ( a ) shows almost no transfected cells. The schematic sketches of a and b are shown in c and d , respectively. e and f show confocal images of nNOS-IR of the same section as a and b , respectively. Note that the staining pattern and distribution of nNOS-IR in the NTS and in adjacent areas in the injected side ( f ) are similar to that of the contralateral side ( e ). Abbreviation : Tr, tractus solitarius. Scale bar = 100 μm

Techniques Used: Injection, Staining, Transfection

24) Product Images from "Feline Immunodeficiency Virus as a Gene Transfer Vector in the Rat Nucleus Tractus Solitarii"

Article Title: Feline Immunodeficiency Virus as a Gene Transfer Vector in the Rat Nucleus Tractus Solitarii

Journal: Cellular and molecular neurobiology

doi: 10.1007/s10571-009-9456-5

Confocal images ( in gray scale ) of the ipsilateral NG with immunofluorescent staining for β Gal and PGP9.5 after FIVLacZ or AdLacZ had been injected unilaterally into the NTS. Neither cells nor fibers labeled for β Gal-IR are seen in the ipsilateral NG after FIVLacZ had been injected to the NTS, as shown in a. c is a confocal image of PGP9.5-IR of the same section to show the structure of the NG section in a. b is a confocal image of NG showing cells (indicated by arrows ) and fibers expressing β Gal-IR after injection of AdLacZ into the NTS. d is confocal image of PGP9.5 showing some NG cells ( arrows ) that are double labeled for β Gal-IR and PGP9.5-IR. Scale bar = 100 μm
Figure Legend Snippet: Confocal images ( in gray scale ) of the ipsilateral NG with immunofluorescent staining for β Gal and PGP9.5 after FIVLacZ or AdLacZ had been injected unilaterally into the NTS. Neither cells nor fibers labeled for β Gal-IR are seen in the ipsilateral NG after FIVLacZ had been injected to the NTS, as shown in a. c is a confocal image of PGP9.5-IR of the same section to show the structure of the NG section in a. b is a confocal image of NG showing cells (indicated by arrows ) and fibers expressing β Gal-IR after injection of AdLacZ into the NTS. d is confocal image of PGP9.5 showing some NG cells ( arrows ) that are double labeled for β Gal-IR and PGP9.5-IR. Scale bar = 100 μm

Techniques Used: Staining, Injection, Labeling, Expressing

Pseudocolored confocal images of rat NTS after FIVLacZ ( a–f ) had been injected into this nucleus, which was then subjected to double-label immunofluorescent staining for β Gal ( green ) and NeuN ( red ), or for β Gal ( green ) and GFAP ( red ), or for β Gal ( green ) and nNOS ( red ). c is a merged confocal image of a (FIVLacZ transfected, green) and b (NeuN-IR, red ). White arrows in a–c indicate representative cells that are stained for both β Gal-IR and NeuN-IR and thus appear yellow in the merged image. Note that the majority of FIVLacZ transfected NTS cells also contained NeuN-IR. f is the merged confocal image of d (FIVLacZ transfected, green ) and e (GFAP-IR, red ). Blue arrows in d and f indicate cells that are positive for β Gal-IR alone. Empty white arrows in e and f indicate cells or processes that are positive for GFAP-IR alone. The majority of FIVLacZ-transfected NTS cells do not contain GFAP-IR. i is a merged confocal image of G (FIVLacZ transfected, green ) and h (nNOS-IR, red ). White arrows in g–i indicate cells that are stained for both β Gal-IR and nNOS-IR. Scale bar = 20 μm
Figure Legend Snippet: Pseudocolored confocal images of rat NTS after FIVLacZ ( a–f ) had been injected into this nucleus, which was then subjected to double-label immunofluorescent staining for β Gal ( green ) and NeuN ( red ), or for β Gal ( green ) and GFAP ( red ), or for β Gal ( green ) and nNOS ( red ). c is a merged confocal image of a (FIVLacZ transfected, green) and b (NeuN-IR, red ). White arrows in a–c indicate representative cells that are stained for both β Gal-IR and NeuN-IR and thus appear yellow in the merged image. Note that the majority of FIVLacZ transfected NTS cells also contained NeuN-IR. f is the merged confocal image of d (FIVLacZ transfected, green ) and e (GFAP-IR, red ). Blue arrows in d and f indicate cells that are positive for β Gal-IR alone. Empty white arrows in e and f indicate cells or processes that are positive for GFAP-IR alone. The majority of FIVLacZ-transfected NTS cells do not contain GFAP-IR. i is a merged confocal image of G (FIVLacZ transfected, green ) and h (nNOS-IR, red ). White arrows in g–i indicate cells that are stained for both β Gal-IR and nNOS-IR. Scale bar = 20 μm

Techniques Used: Injection, Staining, Transfection

Confocal images ( in gray scale ) of the rat NTS after FIVLacZ had been injected unilaterally into this nucleus, which then was subjected to immunofluorescent staining for β Gal ( a, b ) and nNOS ( e, f ). Numerous cells positive for β Gal-IR are observed in the injected side ( b ). A few transfected cells in the adjacent areas, such as the dorsal motor nucleus of vagus (DMV), area postrema (AP), and gracilus nucleus (Gr) were also noted. The contralateral NTS ( a ) shows almost no transfected cells. The schematic sketches of a and b are shown in c and d , respectively. e and f show confocal images of nNOS-IR of the same section as a and b , respectively. Note that the staining pattern and distribution of nNOS-IR in the NTS and in adjacent areas in the injected side ( f ) are similar to that of the contralateral side ( e ). Abbreviation : Tr, tractus solitarius. Scale bar = 100 μm
Figure Legend Snippet: Confocal images ( in gray scale ) of the rat NTS after FIVLacZ had been injected unilaterally into this nucleus, which then was subjected to immunofluorescent staining for β Gal ( a, b ) and nNOS ( e, f ). Numerous cells positive for β Gal-IR are observed in the injected side ( b ). A few transfected cells in the adjacent areas, such as the dorsal motor nucleus of vagus (DMV), area postrema (AP), and gracilus nucleus (Gr) were also noted. The contralateral NTS ( a ) shows almost no transfected cells. The schematic sketches of a and b are shown in c and d , respectively. e and f show confocal images of nNOS-IR of the same section as a and b , respectively. Note that the staining pattern and distribution of nNOS-IR in the NTS and in adjacent areas in the injected side ( f ) are similar to that of the contralateral side ( e ). Abbreviation : Tr, tractus solitarius. Scale bar = 100 μm

Techniques Used: Injection, Staining, Transfection

25) Product Images from "The safety and longevity of DNA vaccines for fish"

Article Title: The safety and longevity of DNA vaccines for fish

Journal: Immunology

doi: 10.1046/j.1365-2567.1999.00688.x

Counting of β-gal-positive muscle fibres at different times after intramuscular administration of pCMV-lacZ. Counts from the control fish were negative and are not shown.
Figure Legend Snippet: Counting of β-gal-positive muscle fibres at different times after intramuscular administration of pCMV-lacZ. Counts from the control fish were negative and are not shown.

Techniques Used: Fluorescence In Situ Hybridization

26) Product Images from "Priming of a ?-Galactosidase (?-GAL)-Specific Type 1 Response in BALB/c Mice Infected with ?-GAL-Transfected Leishmania major"

Article Title: Priming of a ?-Galactosidase (?-GAL)-Specific Type 1 Response in BALB/c Mice Infected with ?-GAL-Transfected Leishmania major

Journal: Infection and Immunity

doi:

L. major-NEO induces only a leishmanial-specific response in BALB/c and C3H mice. BALB/c mice (A) and C3H mice (B) were injected with 2.5 × 10 6 L. major-NEO amastigotes s.c. in one hind footpad. At the indicated times thereafter, the draining LNC were plated at the rate of 4 × 10 5 per well. The LNC were stimulated with L. major (10 6 /ml) or β-GAL (100 μg/ml). The degree of proliferation was assessed by scintillation counting; see Materials and Methods for detailed techniques. Results are from triplicate wells (means ± standard deviations) and are representative of three independent experiments. ∗, no proliferation detected.
Figure Legend Snippet: L. major-NEO induces only a leishmanial-specific response in BALB/c and C3H mice. BALB/c mice (A) and C3H mice (B) were injected with 2.5 × 10 6 L. major-NEO amastigotes s.c. in one hind footpad. At the indicated times thereafter, the draining LNC were plated at the rate of 4 × 10 5 per well. The LNC were stimulated with L. major (10 6 /ml) or β-GAL (100 μg/ml). The degree of proliferation was assessed by scintillation counting; see Materials and Methods for detailed techniques. Results are from triplicate wells (means ± standard deviations) and are representative of three independent experiments. ∗, no proliferation detected.

Techniques Used: Mouse Assay, Injection

L. major-βGAL infected Mφs do not activate β-GAL-specific T cells unless the Mφs are activated with IFN-γ plus LPS. Mφs were cultured with (i) soluble β-GAL or (ii) either L. major-βGAL ( Lm-βGAL ) or L. major-NEO ( Lm-NEO ) (indicated infection rates, 0.5 parasite per Mφ, 1 parasite per Mφ, etc.). After rinsing away β-GAL or nonphagocytized parasites, the β-GAL-specific, IL-2-secreting T-cell hybridoma, 1E3.03.H4, was added. The degree of activation of 1E3.03.H4 was determined by measuring the levels of IL-2 in the culture supernatants. For details of these techniques, see Materials and Methods. Results are representative of four independent experiments. ∗, no IL-2 detected. To ensure that the results obtained were not due to differences in the uptake of L. major-βGAL ) at the end of the experiments when supernatants were harvested for IL-2 testing. Unstimulated Mφs (infected with a ratio of 2 parasites/Mφ) contained 170 ± 41 (mean ± standard deviation) intracellular L. major-βGAL per 100 Mφs, while IFN-γ plus LPS-treated Mφs contained 121 ± 21 parasites.
Figure Legend Snippet: L. major-βGAL infected Mφs do not activate β-GAL-specific T cells unless the Mφs are activated with IFN-γ plus LPS. Mφs were cultured with (i) soluble β-GAL or (ii) either L. major-βGAL ( Lm-βGAL ) or L. major-NEO ( Lm-NEO ) (indicated infection rates, 0.5 parasite per Mφ, 1 parasite per Mφ, etc.). After rinsing away β-GAL or nonphagocytized parasites, the β-GAL-specific, IL-2-secreting T-cell hybridoma, 1E3.03.H4, was added. The degree of activation of 1E3.03.H4 was determined by measuring the levels of IL-2 in the culture supernatants. For details of these techniques, see Materials and Methods. Results are representative of four independent experiments. ∗, no IL-2 detected. To ensure that the results obtained were not due to differences in the uptake of L. major-βGAL ) at the end of the experiments when supernatants were harvested for IL-2 testing. Unstimulated Mφs (infected with a ratio of 2 parasites/Mφ) contained 170 ± 41 (mean ± standard deviation) intracellular L. major-βGAL per 100 Mφs, while IFN-γ plus LPS-treated Mφs contained 121 ± 21 parasites.

Techniques Used: Infection, Cell Culture, Activation Assay, Standard Deviation

27) Product Images from "A robust lentiviral pseudotype neutralisation assay for in-field serosurveillance of rabies and lyssaviruses in Africa"

Article Title: A robust lentiviral pseudotype neutralisation assay for in-field serosurveillance of rabies and lyssaviruses in Africa

Journal: Vaccine

doi: 10.1016/j.vaccine.2009.09.024

Repertoire of reporter genes that can be carried within pseudotypes. We have expanded the range of reporter genes to enable the neutralisation assay to be performed in a much greater number of laboratories than previously possible. LacZ -based pseudotypes can be detected with X-gal, CPRG or ONPG β-gal substrates that turn blue, red or yellow in the presence of the enzyme, respectively. GFP appears as green cytoplasmic staining while the oxidation of the luciferase substrate results in light emission that can be detected in a luminometer. [VNAb]: virus-neutralising antibody concentration. The arrow indicates zero luciferase activity.
Figure Legend Snippet: Repertoire of reporter genes that can be carried within pseudotypes. We have expanded the range of reporter genes to enable the neutralisation assay to be performed in a much greater number of laboratories than previously possible. LacZ -based pseudotypes can be detected with X-gal, CPRG or ONPG β-gal substrates that turn blue, red or yellow in the presence of the enzyme, respectively. GFP appears as green cytoplasmic staining while the oxidation of the luciferase substrate results in light emission that can be detected in a luminometer. [VNAb]: virus-neutralising antibody concentration. The arrow indicates zero luciferase activity.

Techniques Used: Staining, Luciferase, Concentration Assay, Activity Assay

28) Product Images from "Novel Fe3+-Based 1H MRI β-Galactosidase Reporter Molecules**"

Article Title: Novel Fe3+-Based 1H MRI β-Galactosidase Reporter Molecules**

Journal: ChemPlusChem

doi: 10.1002/cplu.201100072

T 1 and T 2 response to β-gal
Figure Legend Snippet: T 1 and T 2 response to β-gal

Techniques Used:

29) Product Images from "Mouse Horizontal Cells do not Express Connexin26 or Connexin36"

Article Title: Mouse Horizontal Cells do not Express Connexin26 or Connexin36

Journal: Cell communication & adhesion

doi:

Cx26 immunopositive puncta (green) could not be detected on calbindin positive processes(red) in the OPL( A ), but could be easily located in control sections from mouse liver ( B ) counterstained with the nuclear label DAPI (blue). In retina from mice in which one copy of Cx36 was replaced with β-gal, immunofluorescence for this reporter (green) does not colocalize with calbindin (red) positive horizontal cells ( C ). ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer. Arrows indicate horizontal cell bodies.
Figure Legend Snippet: Cx26 immunopositive puncta (green) could not be detected on calbindin positive processes(red) in the OPL( A ), but could be easily located in control sections from mouse liver ( B ) counterstained with the nuclear label DAPI (blue). In retina from mice in which one copy of Cx36 was replaced with β-gal, immunofluorescence for this reporter (green) does not colocalize with calbindin (red) positive horizontal cells ( C ). ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer. Arrows indicate horizontal cell bodies.

Techniques Used: Mouse Assay, Immunofluorescence

30) Product Images from "Human Vascular Endothelial Growth Factor A165 Expression Induces the Mouse Model of Neovascular Age-Related Macular Degeneration"

Article Title: Human Vascular Endothelial Growth Factor A165 Expression Induces the Mouse Model of Neovascular Age-Related Macular Degeneration

Journal: Genes

doi: 10.3390/genes9090438

Morphologic changes and transgene expression in the Cre and LacZ injected eyes. ( a ) β-gal expression (arrowhead) after LacZ injection was seen two weeks after gene transfer but not at later time points. ( b ) VEGF-A expression (violet) in ganglion cell layer (black arrowhead), photoreceptors (arrowhead), and neovascular membrane (arrow) in the eye of Cre -injected mouse. ( c ) Glial fibrillary acidic protein (GFAP) immunoreactivity was observed in the nerve fiber layer (arrowhead) and Müller cells (arrow) in the outer retina post- Cre injection. ( d ) In the Cre group, F4/80 positive macrophages were seen in the retina and subretinal layers. ( e , f ) Retinal autofluorescence (yellow) with DAPI nuclear counterstain (blue). In Cre -injected retina (e), drusen-like lipofuscin deposits (arrowhead) and the loss of photoreceptors were seen. Intact photoreceptor layer (f, arrowhead) was observed in LacZ -injected eyes. Scale bar is 100 µm. GCL: Ganglion cell layer.
Figure Legend Snippet: Morphologic changes and transgene expression in the Cre and LacZ injected eyes. ( a ) β-gal expression (arrowhead) after LacZ injection was seen two weeks after gene transfer but not at later time points. ( b ) VEGF-A expression (violet) in ganglion cell layer (black arrowhead), photoreceptors (arrowhead), and neovascular membrane (arrow) in the eye of Cre -injected mouse. ( c ) Glial fibrillary acidic protein (GFAP) immunoreactivity was observed in the nerve fiber layer (arrowhead) and Müller cells (arrow) in the outer retina post- Cre injection. ( d ) In the Cre group, F4/80 positive macrophages were seen in the retina and subretinal layers. ( e , f ) Retinal autofluorescence (yellow) with DAPI nuclear counterstain (blue). In Cre -injected retina (e), drusen-like lipofuscin deposits (arrowhead) and the loss of photoreceptors were seen. Intact photoreceptor layer (f, arrowhead) was observed in LacZ -injected eyes. Scale bar is 100 µm. GCL: Ganglion cell layer.

Techniques Used: Expressing, Injection

31) Product Images from "The safety and longevity of DNA vaccines for fish"

Article Title: The safety and longevity of DNA vaccines for fish

Journal: Immunology

doi: 10.1046/j.1365-2567.1999.00688.x

Counting of β-gal-positive muscle fibres at different times after intramuscular administration of pCMV-lacZ. Counts from the control fish were negative and are not shown.
Figure Legend Snippet: Counting of β-gal-positive muscle fibres at different times after intramuscular administration of pCMV-lacZ. Counts from the control fish were negative and are not shown.

Techniques Used: Fluorescence In Situ Hybridization

32) Product Images from "Perinuclear localization of slow troponin C m RNA in muscle cells is controlled by a cis-element located at its 3? untranslated region"

Article Title: Perinuclear localization of slow troponin C m RNA in muscle cells is controlled by a cis-element located at its 3? untranslated region

Journal: RNA

doi: 10.1261/rna.5460105

Regulation of cytoplasmic distribution of β-Gal mRNA by the 3′ UTR of sTnC mRNA in differentiated muscle cells. Differentiating C 2 C 12 cells were transfected with different β-Gal constructs as described in Materials and Methods.
Figure Legend Snippet: Regulation of cytoplasmic distribution of β-Gal mRNA by the 3′ UTR of sTnC mRNA in differentiated muscle cells. Differentiating C 2 C 12 cells were transfected with different β-Gal constructs as described in Materials and Methods.

Techniques Used: Transfection, Construct

Cell specificity of sTnC mRNA localization signal. Proliferating C 2 C 12 myoblasts and HeLa cells were transfected with pCMV-SPORT-β-Gal or construct J containing the sTnC mRNAs 40-nt-long localization signal. Twenty-four hours after transfection,
Figure Legend Snippet: Cell specificity of sTnC mRNA localization signal. Proliferating C 2 C 12 myoblasts and HeLa cells were transfected with pCMV-SPORT-β-Gal or construct J containing the sTnC mRNAs 40-nt-long localization signal. Twenty-four hours after transfection,

Techniques Used: Transfection, Construct

Distribution of β-Gal mRNA in transiently transfected C 2 C 12 myocytes with β-gal-sTnC chimeras. These are schematic representations (not drawn to scale) of the different constructs used for β-Gal distribution studies. The CMV promoter
Figure Legend Snippet: Distribution of β-Gal mRNA in transiently transfected C 2 C 12 myocytes with β-gal-sTnC chimeras. These are schematic representations (not drawn to scale) of the different constructs used for β-Gal distribution studies. The CMV promoter

Techniques Used: Transfection, Construct

Levels of β-Gal polypeptide and mRNA in cells transfected with different β-gal-sTnC chimeric constructs. Differentiating C 2 C 12 cells were cotransfected with a β-gal-sTnC chimeric construct and a GFP expression vector. Twenty-four
Figure Legend Snippet: Levels of β-Gal polypeptide and mRNA in cells transfected with different β-gal-sTnC chimeric constructs. Differentiating C 2 C 12 cells were cotransfected with a β-gal-sTnC chimeric construct and a GFP expression vector. Twenty-four

Techniques Used: Transfection, Construct, Expressing, Plasmid Preparation

33) Product Images from "Variant allele frequency enrichment analysis in vitro reveals sonic hedgehog pathway to impede sustained temozolomide response in GBM"

Article Title: Variant allele frequency enrichment analysis in vitro reveals sonic hedgehog pathway to impede sustained temozolomide response in GBM

Journal: Scientific Reports

doi: 10.1038/srep07915

(A) Light microscopic images (50 μm bar) at single cellular level. ( a ), day 5 DMSO-treated control cells of A49910, ( b ), day 28 DMSO-treated control cells of A49910. ( c ), day 5 DMSO-treated control cells of M45481, (d), day 28 DMSO-treated control cells of M4548, ( e ), day 5 TMZ-treated cells of A49910, ( f ), day 28 post-TMZ-treated cells of A49910, ( g ), day 5 TMZ-treated cells of M45481, and ( h ), day 28 post-TMZ-treated cells of M45481. ( B ) Light microscopic images (50 μm bar) of the cells showing SA-β-Gal staining following TMZ treatment and post-treatment recovery. ( a ), day 5 DMSO-treated control cells of A49910, ( b ), day 28 DMSO-treated control cells of A49910. ( c ), day 5 DMSO-treated control cells of M45481, ( d ), day 28 DMSO-treated control cells of M4548, ( e ), day 5 TMZ-treated cells of A49910, ( f ), day 28 post-TMZ-treated cells of A49910, ( g ), day 5 TMZ-treated cells of M45481, and ( h ), day 28 post-TMZ-treated cells of M45481. ( C ) VAFs showing biphasic trends where the first phase is the comparison of VAFs between C5 to T5 and second phase is the comparison of VAFs between T5 to T28. Red arrows are showing enriching VAFs in upward direction, blue showing downward direction and black showing no significant change. The green and purple horizontal bars on the right side represent the number of genes (N) in each category in A49910 and in M45481 respectively. ( D ) Sanger sequencing chromatogram showing a G to A transition on STK36 gene (arrow mark) in M45481 (lower panel) but not in A49910 (upper panel). ( E ) Showing mRNA expression patterns of Hh-pathway component genes, STK36 ( a ), GLI1 ( b ), GLI2 ( c ), GLI3 ( d ), Hh-pathway target gene SNAI1 ( e ) and MGMT ( f ) following TMZ-treatment and post-treatment recovery (C5, DMSO treated control; T5, day-5 TMZ-treated; T28, day-28 post-treatment recovery, ✶ p-value
Figure Legend Snippet: (A) Light microscopic images (50 μm bar) at single cellular level. ( a ), day 5 DMSO-treated control cells of A49910, ( b ), day 28 DMSO-treated control cells of A49910. ( c ), day 5 DMSO-treated control cells of M45481, (d), day 28 DMSO-treated control cells of M4548, ( e ), day 5 TMZ-treated cells of A49910, ( f ), day 28 post-TMZ-treated cells of A49910, ( g ), day 5 TMZ-treated cells of M45481, and ( h ), day 28 post-TMZ-treated cells of M45481. ( B ) Light microscopic images (50 μm bar) of the cells showing SA-β-Gal staining following TMZ treatment and post-treatment recovery. ( a ), day 5 DMSO-treated control cells of A49910, ( b ), day 28 DMSO-treated control cells of A49910. ( c ), day 5 DMSO-treated control cells of M45481, ( d ), day 28 DMSO-treated control cells of M4548, ( e ), day 5 TMZ-treated cells of A49910, ( f ), day 28 post-TMZ-treated cells of A49910, ( g ), day 5 TMZ-treated cells of M45481, and ( h ), day 28 post-TMZ-treated cells of M45481. ( C ) VAFs showing biphasic trends where the first phase is the comparison of VAFs between C5 to T5 and second phase is the comparison of VAFs between T5 to T28. Red arrows are showing enriching VAFs in upward direction, blue showing downward direction and black showing no significant change. The green and purple horizontal bars on the right side represent the number of genes (N) in each category in A49910 and in M45481 respectively. ( D ) Sanger sequencing chromatogram showing a G to A transition on STK36 gene (arrow mark) in M45481 (lower panel) but not in A49910 (upper panel). ( E ) Showing mRNA expression patterns of Hh-pathway component genes, STK36 ( a ), GLI1 ( b ), GLI2 ( c ), GLI3 ( d ), Hh-pathway target gene SNAI1 ( e ) and MGMT ( f ) following TMZ-treatment and post-treatment recovery (C5, DMSO treated control; T5, day-5 TMZ-treated; T28, day-28 post-treatment recovery, ✶ p-value

Techniques Used: Staining, Sequencing, Expressing

34) Product Images from "Regulation of Drug Reward by cAMP Response Element-Binding Protein: Evidence for Two Functionally Distinct Subregions of the Ventral Tegmental Area"

Article Title: Regulation of Drug Reward by cAMP Response Element-Binding Protein: Evidence for Two Functionally Distinct Subregions of the Ventral Tegmental Area

Journal: The Journal of Neuroscience

doi: 10.1523/JNEUROSCI.0345-05.2005

A large portion of neurons infected in caudal VTA project to the NAc and are dopaminergic. The number of β-gal-positive neurons did not significantly differ between rostral and caudal VTA. However, more neurons were retrogradely labeled in caudal VTA than in rostral VTA after injection of HS in to the NAc. More neurons in caudal VTA than in rostral VTA also were colabeled with β-gal and HS. Most of the HS- and β-gal-positive neurons in rostral and caudal VTA were also TH positive. B-G , A representative section from caudal VTA, triple labeled for HS ( B ), β-gal ( C ), and TH ( D ). E , Merged image from B and C showing neurons colabeled with HS and β-gal. F , Merged image from B and D showing neurons colabeled with HS and TH. G , Merged image from B-D showing triple-labeled neurons (arrows). Data are expressed as cell numbers per section ± SEM ( n = 3). * p
Figure Legend Snippet: A large portion of neurons infected in caudal VTA project to the NAc and are dopaminergic. The number of β-gal-positive neurons did not significantly differ between rostral and caudal VTA. However, more neurons were retrogradely labeled in caudal VTA than in rostral VTA after injection of HS in to the NAc. More neurons in caudal VTA than in rostral VTA also were colabeled with β-gal and HS. Most of the HS- and β-gal-positive neurons in rostral and caudal VTA were also TH positive. B-G , A representative section from caudal VTA, triple labeled for HS ( B ), β-gal ( C ), and TH ( D ). E , Merged image from B and C showing neurons colabeled with HS and β-gal. F , Merged image from B and D showing neurons colabeled with HS and TH. G , Merged image from B-D showing triple-labeled neurons (arrows). Data are expressed as cell numbers per section ± SEM ( n = 3). * p

Techniques Used: Infection, Labeling, Injection

35) Product Images from "Promyelinating Schwann Cells Express Tst-1/SCIP/Oct-6"

Article Title: Promyelinating Schwann Cells Express Tst-1/SCIP/Oct-6

Journal: The Journal of Neuroscience

doi: 10.1523/JNEUROSCI.18-19-07891.1998

Solution assay of β-gal in adult tst-1/scip/oct-6 +/− sciatic nerves. The heights of the bars represent the mean RLUs per milligram of protein from the distal stumps of nerves 1, 4, 8, 12, 24, and 58 d after transection or after crushing. Note that the RLUs of the crushed nerves is higher than that in the transected nerves during the period of ensheathment and myelination (8–24 d after crushing).
Figure Legend Snippet: Solution assay of β-gal in adult tst-1/scip/oct-6 +/− sciatic nerves. The heights of the bars represent the mean RLUs per milligram of protein from the distal stumps of nerves 1, 4, 8, 12, 24, and 58 d after transection or after crushing. Note that the RLUs of the crushed nerves is higher than that in the transected nerves during the period of ensheathment and myelination (8–24 d after crushing).

Techniques Used:

Solution assay of β-gal in developing tst-1/scip/oct-6 +/− sciatic nerves. The heights of the bars represent the mean RLUs per milligram of protein for P1, P5, P10, P15, P20, P37, and adult mouse sciatic nerves. The RLUs were measured in triplicate for each sample; error bars represent the SEM.
Figure Legend Snippet: Solution assay of β-gal in developing tst-1/scip/oct-6 +/− sciatic nerves. The heights of the bars represent the mean RLUs per milligram of protein for P1, P5, P10, P15, P20, P37, and adult mouse sciatic nerves. The RLUs were measured in triplicate for each sample; error bars represent the SEM.

Techniques Used:

36) Product Images from "Effects of in vivo adventitial expression of recombinant endothelial nitric oxide synthase gene in cerebral arteries"

Article Title: Effects of in vivo adventitial expression of recombinant endothelial nitric oxide synthase gene in cerebral arteries

Journal: Proceedings of the National Academy of Sciences of the United States of America

doi:

β-Gal levels in AdCMVLacZ- and AdCMVNOS-transduced cerebral arteries 24 h after gene transfer. ( A ) β-Gal levels in basilar and middle cerebral arteries of same dogs after in vivo gene transfer (final viral titer in CSF, 10 9 pfu/ml). ( B ) β-Gal levels in basilar arteries of same dogs after ex vivo gene transfer (final viral titers in the incubation media, 10 8 , 10 9 , and 10 10 pfu/ml). Data are expressed as means ± SEM ( n = 4 from 4 dogs). ∗, P
Figure Legend Snippet: β-Gal levels in AdCMVLacZ- and AdCMVNOS-transduced cerebral arteries 24 h after gene transfer. ( A ) β-Gal levels in basilar and middle cerebral arteries of same dogs after in vivo gene transfer (final viral titer in CSF, 10 9 pfu/ml). ( B ) β-Gal levels in basilar arteries of same dogs after ex vivo gene transfer (final viral titers in the incubation media, 10 8 , 10 9 , and 10 10 pfu/ml). Data are expressed as means ± SEM ( n = 4 from 4 dogs). ∗, P

Techniques Used: In Vivo, Ex Vivo, Incubation

Morphological demonstration of transgene expression on canine brain and cerebral arteries 24 h after intracisternal injection of viral vector (final viral titer in CSF, 10 9 pfu/ml). ( A ) β-Gal staining on the dorsal surface of the brain. ( B ) β-Gal staining on the ventral surface of the same brain. Note increased β-gal staining in B compared with A . ( C ) Microscopic view (cross section) of β-gal staining in a basilar artery after being counterstained with nuclear fast red. ( D ) Microscopic view (cross section) of immunohistochemical staining of eNOS in a middle cerebral artery 24 h after in vivo eNOS gene transfer. (Bar = 0.1 mm.)
Figure Legend Snippet: Morphological demonstration of transgene expression on canine brain and cerebral arteries 24 h after intracisternal injection of viral vector (final viral titer in CSF, 10 9 pfu/ml). ( A ) β-Gal staining on the dorsal surface of the brain. ( B ) β-Gal staining on the ventral surface of the same brain. Note increased β-gal staining in B compared with A . ( C ) Microscopic view (cross section) of β-gal staining in a basilar artery after being counterstained with nuclear fast red. ( D ) Microscopic view (cross section) of immunohistochemical staining of eNOS in a middle cerebral artery 24 h after in vivo eNOS gene transfer. (Bar = 0.1 mm.)

Techniques Used: Expressing, Injection, Plasmid Preparation, Staining, Immunohistochemistry, In Vivo

37) Product Images from "Monitoring protein-protein interactions in intact eukaryotic cells by ?-galactosidase complementation"

Article Title: Monitoring protein-protein interactions in intact eukaryotic cells by ?-galactosidase complementation

Journal: Proceedings of the National Academy of Sciences of the United States of America

doi:

Biochemical assay of induction of β-gal activity upon chimera complementation. ( A ) Kinetics of induction of β-gal activity upon treatment with rapamycin. Pure populations of C2C12 cells stably expressing both FKBP12-Δω and FRAP-Δα were plated in 96-well plates and 10 ng/ml rapamycin was added at time zero. Cells were then lysed at different time intervals thereafter, and the β-gal activity in the lysates was quantitated by chemiluminescence. ( B ) Dose response of β-gal activity upon rapamycin treatment. C2C12 cells expressing both FKBP12-Δω and FRAP-Δα were plated in 96-well plates and treated with different concentrations of rapamycin for 3.5 hr. β-gal activity is expressed as luminescence counts per second. Each point represents the average of six replicate samples. Error bars indicate standard deviations from the mean.
Figure Legend Snippet: Biochemical assay of induction of β-gal activity upon chimera complementation. ( A ) Kinetics of induction of β-gal activity upon treatment with rapamycin. Pure populations of C2C12 cells stably expressing both FKBP12-Δω and FRAP-Δα were plated in 96-well plates and 10 ng/ml rapamycin was added at time zero. Cells were then lysed at different time intervals thereafter, and the β-gal activity in the lysates was quantitated by chemiluminescence. ( B ) Dose response of β-gal activity upon rapamycin treatment. C2C12 cells expressing both FKBP12-Δω and FRAP-Δα were plated in 96-well plates and treated with different concentrations of rapamycin for 3.5 hr. β-gal activity is expressed as luminescence counts per second. Each point represents the average of six replicate samples. Error bars indicate standard deviations from the mean.

Techniques Used: Activity Assay, Stable Transfection, Expressing

Experimental design. ( A ) When the Δα and Δω β-gal mutants are fused to proteins that do not dimerize, their association is not favored and β-gal activity is not detected. ( B ) When the Δα and Δω β-gal mutants are fused to proteins that can dimerize, the formation of active β-gal is favored. ( C ) Schematic representation of the FKBP12-Δω-Neo and the FRAP-Δα-Hygro constructs. IRES, internal ribosome entry sequence; LTR, long terminal repeat.
Figure Legend Snippet: Experimental design. ( A ) When the Δα and Δω β-gal mutants are fused to proteins that do not dimerize, their association is not favored and β-gal activity is not detected. ( B ) When the Δα and Δω β-gal mutants are fused to proteins that can dimerize, the formation of active β-gal is favored. ( C ) Schematic representation of the FKBP12-Δω-Neo and the FRAP-Δα-Hygro constructs. IRES, internal ribosome entry sequence; LTR, long terminal repeat.

Techniques Used: Activity Assay, Construct, Sequencing

Histochemical assay of induction of β-gal activity upon chimera complementation. C2C12 cells expressing both FKBP12-Δω and FRAP-Δα were maintained overnight either in the absence ( A and C ) or the presence ( B and D ) of 10 ng/ml rapamycin. β-gal activity was visualized by fluorescence microscopy using Fluor-X-Gal as substrate. ( A and B ) Double-labeled samples showing Hoechst stained nuclei (blue) and β-gal activity using Fluor-X-Gal as substrate viewed with a rhodamine filter set (red). ( C and D ) Triple-labeled samples obtained by imaging with a DeltaVision microscope showing β-gal activity using Fluor-X-Gal as substrate (green), Hoechst stained nuclei (blue), and Cy5-labeled actin filaments (red) to visualize the contour of each cell at higher magnification.
Figure Legend Snippet: Histochemical assay of induction of β-gal activity upon chimera complementation. C2C12 cells expressing both FKBP12-Δω and FRAP-Δα were maintained overnight either in the absence ( A and C ) or the presence ( B and D ) of 10 ng/ml rapamycin. β-gal activity was visualized by fluorescence microscopy using Fluor-X-Gal as substrate. ( A and B ) Double-labeled samples showing Hoechst stained nuclei (blue) and β-gal activity using Fluor-X-Gal as substrate viewed with a rhodamine filter set (red). ( C and D ) Triple-labeled samples obtained by imaging with a DeltaVision microscope showing β-gal activity using Fluor-X-Gal as substrate (green), Hoechst stained nuclei (blue), and Cy5-labeled actin filaments (red) to visualize the contour of each cell at higher magnification.

Techniques Used: Activity Assay, Expressing, Fluorescence, Microscopy, Labeling, Staining, Imaging

FACS analysis of induced β-gal activity upon chimera complementation. The red peaks represent the untreated samples and the blue peaks represent samples treated with 10 ng/ml rapamycin. ( A ) Induction of β-gal activity in a population of C2C12 cells expressing both FKBP12-Δω and FRAP-Δα after 90 min of rapamycin treatment. The majority of the cells respond to rapamycin treatment with an increase in β-gal activity. ( B ) Subpopulation of cells selected on the basis of low β-gal activity in uninduced conditions. ( C ) The same population was maintained overnight in the absence (red peak) or in the presence (blue peak) of rapamycin. The induced and uninduced populations yield essentially nonoverlapping peaks. The vertical axis represents relative cell number and the horizontal axis represents intensity of β-gal fluorescence on a logarithmic scale.
Figure Legend Snippet: FACS analysis of induced β-gal activity upon chimera complementation. The red peaks represent the untreated samples and the blue peaks represent samples treated with 10 ng/ml rapamycin. ( A ) Induction of β-gal activity in a population of C2C12 cells expressing both FKBP12-Δω and FRAP-Δα after 90 min of rapamycin treatment. The majority of the cells respond to rapamycin treatment with an increase in β-gal activity. ( B ) Subpopulation of cells selected on the basis of low β-gal activity in uninduced conditions. ( C ) The same population was maintained overnight in the absence (red peak) or in the presence (blue peak) of rapamycin. The induced and uninduced populations yield essentially nonoverlapping peaks. The vertical axis represents relative cell number and the horizontal axis represents intensity of β-gal fluorescence on a logarithmic scale.

Techniques Used: FACS, Activity Assay, Expressing, Fluorescence

38) Product Images from "S-Gal®, A Novel 1H MRI Reporter for ?-Galactosidase"

Article Title: S-Gal®, A Novel 1H MRI Reporter for ?-Galactosidase

Journal: Magnetic resonance in medicine : official journal of the Society of Magnetic Resonance in Medicine / Society of Magnetic Resonance in Medicine

doi: 10.1002/mrm.22400

MRI detection of beta β-gal activity in stably transfected breast tumors
Figure Legend Snippet: MRI detection of beta β-gal activity in stably transfected breast tumors

Techniques Used: Magnetic Resonance Imaging, Activity Assay, Stable Transfection, Transfection

Relaxation studies with various FAC and β-galactosidase concentrations
Figure Legend Snippet: Relaxation studies with various FAC and β-galactosidase concentrations

Techniques Used:

39) Product Images from "Progenitor Cell Capacity of NeuroD1-Expressing Globose Basal Cells in the Mouse Olfactory Epithelium"

Article Title: Progenitor Cell Capacity of NeuroD1-Expressing Globose Basal Cells in the Mouse Olfactory Epithelium

Journal: The Journal of comparative neurology

doi: 10.1002/cne.22726

NeuroD1 is expressed in the vast majority of OSNs at some point during their differentiation. A–C: Sections from the olfactory epithelium of adult, Δ NeuroD1-Cre × Rosa26-flox-stop-flox-LacZ mice are stained with β-gal and
Figure Legend Snippet: NeuroD1 is expressed in the vast majority of OSNs at some point during their differentiation. A–C: Sections from the olfactory epithelium of adult, Δ NeuroD1-Cre × Rosa26-flox-stop-flox-LacZ mice are stained with β-gal and

Techniques Used: Mouse Assay, Staining

The gustducin(+) solitary chemoreceptor cells apparently do not derive from a NeuroD1-expressing progenitor. Section through a cul-de-sac showing a small patch of olfactory epithelium. A: Anti-β-gal staining marks the cells of the olfactory epithelium
Figure Legend Snippet: The gustducin(+) solitary chemoreceptor cells apparently do not derive from a NeuroD1-expressing progenitor. Section through a cul-de-sac showing a small patch of olfactory epithelium. A: Anti-β-gal staining marks the cells of the olfactory epithelium

Techniques Used: Expressing, Staining

40) Product Images from "Locus Control Region of the Human CD2 Gene in a Lentivirus Vector Confers Position-Independent Transgene Expression"

Article Title: Locus Control Region of the Human CD2 Gene in a Lentivirus Vector Confers Position-Independent Transgene Expression

Journal: Journal of Virology

doi: 10.1128/JVI.75.10.4641-4648.2001

T-cell-specific stimulation of β-Gal expression mediated by the hCD2 LCR in cell pools. HT1080, Jurkat, and primary human T cells were transduced with pHIV/CK-3 or with pHIV/CK-4 at an MOI of 10. Transduced cells were selected with G418. Cell extracts were prepared by freeze-thaw cycles. β-Gal expression in the cell pools was quantified and corrected for the protein concentration of the samples. All assays were carried out at least three times with freshly prepared cell extracts of exponentially growing cells. Standard deviations are indicated.
Figure Legend Snippet: T-cell-specific stimulation of β-Gal expression mediated by the hCD2 LCR in cell pools. HT1080, Jurkat, and primary human T cells were transduced with pHIV/CK-3 or with pHIV/CK-4 at an MOI of 10. Transduced cells were selected with G418. Cell extracts were prepared by freeze-thaw cycles. β-Gal expression in the cell pools was quantified and corrected for the protein concentration of the samples. All assays were carried out at least three times with freshly prepared cell extracts of exponentially growing cells. Standard deviations are indicated.

Techniques Used: Expressing, Transduction, Protein Concentration

LCR-mediated β-Gal gene expression in individual primary T-cell clones. Primary human T cells were transduced with pHIV/CK-3 at an MOI of 10. Transduced cells were selected with G418. The β-Gal activity in individual clones was quantified and corrected for the protein concentration in the samples. The average expression level is indicated by a horizontal line. Each dot represents the average β-Gal activity from three independent assays.
Figure Legend Snippet: LCR-mediated β-Gal gene expression in individual primary T-cell clones. Primary human T cells were transduced with pHIV/CK-3 at an MOI of 10. Transduced cells were selected with G418. The β-Gal activity in individual clones was quantified and corrected for the protein concentration in the samples. The average expression level is indicated by a horizontal line. Each dot represents the average β-Gal activity from three independent assays.

Techniques Used: Expressing, Clone Assay, Transduction, Activity Assay, Protein Concentration

Related Articles

Transduction:

Article Title: Minimum Requirements for Efficient Transduction of Dividing and Nondividing Cells by Feline Immunodeficiency Virus Vectors
Article Snippet: .. Growth-arrested cells were assayed for reporter gene expression 3 days after transduction. β-Gal expression was assayed after the cells were fixed in a solution of 3% formaldehyde and 1.25% glutaraldehyde in PBS and stained for 4 h at 37°C in a solution containing 400 μg of 5-bromo-4-chloro-3-indolyl-β- d -galactopyranoside (X-Gal; Sigma, St. Louis, Mo.)/ml ( ). .. The titer was determined by counting the number of blue foci per well and was reported as LacZ-forming units (LFU) per milliliter of vector stock.

Staining:

Article Title: Minimum Requirements for Efficient Transduction of Dividing and Nondividing Cells by Feline Immunodeficiency Virus Vectors
Article Snippet: .. Growth-arrested cells were assayed for reporter gene expression 3 days after transduction. β-Gal expression was assayed after the cells were fixed in a solution of 3% formaldehyde and 1.25% glutaraldehyde in PBS and stained for 4 h at 37°C in a solution containing 400 μg of 5-bromo-4-chloro-3-indolyl-β- d -galactopyranoside (X-Gal; Sigma, St. Louis, Mo.)/ml ( ). .. The titer was determined by counting the number of blue foci per well and was reported as LacZ-forming units (LFU) per milliliter of vector stock.

Article Title: G-protein stimulatory subunit alpha and Gq/11α G-proteins are both required to maintain quiescent stem-like chondrocytes
Article Snippet: .. β-galactosidase activity was evaluated using X-gal staining, which was performed on either entire bone or unfixed frozen sections using the ‘Stain for Beta-Gal Expression In Tissue’ and ‘Stain for Beta-Gal Expression In Situ’ kits (Millipore) following the manufacturer’s protocol. .. Alizarin red and Alcian blue staining were performed using a modification of McLeod’s method .

Activity Assay:

Article Title: G-protein stimulatory subunit alpha and Gq/11α G-proteins are both required to maintain quiescent stem-like chondrocytes
Article Snippet: .. β-galactosidase activity was evaluated using X-gal staining, which was performed on either entire bone or unfixed frozen sections using the ‘Stain for Beta-Gal Expression In Tissue’ and ‘Stain for Beta-Gal Expression In Situ’ kits (Millipore) following the manufacturer’s protocol. .. Alizarin red and Alcian blue staining were performed using a modification of McLeod’s method .

In Situ:

Article Title: G-protein stimulatory subunit alpha and Gq/11α G-proteins are both required to maintain quiescent stem-like chondrocytes
Article Snippet: .. β-galactosidase activity was evaluated using X-gal staining, which was performed on either entire bone or unfixed frozen sections using the ‘Stain for Beta-Gal Expression In Tissue’ and ‘Stain for Beta-Gal Expression In Situ’ kits (Millipore) following the manufacturer’s protocol. .. Alizarin red and Alcian blue staining were performed using a modification of McLeod’s method .

Expressing:

Article Title: Minimum Requirements for Efficient Transduction of Dividing and Nondividing Cells by Feline Immunodeficiency Virus Vectors
Article Snippet: .. Growth-arrested cells were assayed for reporter gene expression 3 days after transduction. β-Gal expression was assayed after the cells were fixed in a solution of 3% formaldehyde and 1.25% glutaraldehyde in PBS and stained for 4 h at 37°C in a solution containing 400 μg of 5-bromo-4-chloro-3-indolyl-β- d -galactopyranoside (X-Gal; Sigma, St. Louis, Mo.)/ml ( ). .. The titer was determined by counting the number of blue foci per well and was reported as LacZ-forming units (LFU) per milliliter of vector stock.

Article Title: G-protein stimulatory subunit alpha and Gq/11α G-proteins are both required to maintain quiescent stem-like chondrocytes
Article Snippet: .. β-galactosidase activity was evaluated using X-gal staining, which was performed on either entire bone or unfixed frozen sections using the ‘Stain for Beta-Gal Expression In Tissue’ and ‘Stain for Beta-Gal Expression In Situ’ kits (Millipore) following the manufacturer’s protocol. .. Alizarin red and Alcian blue staining were performed using a modification of McLeod’s method .

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  • 86
    Millipore microglial sa β gal activity
    Microglia aged in culture display signs of senescence, including increased senescent-associated β-galactosidase <t>(SA-β-gal)</t> activity and microRNA (miR)-146a expression. Microglial cells were kept in culture for 2 and 16 days in vitro (DIV). Activity of SA-β-gal was determined using a commercial kit. (A) Representative images of 2 and 16 DIV microglia showing SA-β-gal staining. (B) SA-β-gal-positive cells were counted and results expressed in graph bars as mean ± SEM. (C) miR-146a expression was evaluated by Real-Time PCR. Results are expressed in graph bars as mean ± SEM. Cultures, n = 4 per group. t -test, * p
    Microglial Sa β Gal Activity, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Millipore senescence assay senescence associated β galactosidase activity
    Apoptosis and senescence in tumor sections from control and mixed cells. (A) Sections of tumor tissues were subjected to TUNEL assay. Blue colour shows nuclei of cells stained with DAPI. Green foci show TUNEL positive regions. (B) Tumor sections processed for SA-β-gal assay. The part of image is shown below in the respective inset. Greenish blue color indicates active <t>SA-β-galactosidase.</t>
    Senescence Assay Senescence Associated β Galactosidase Activity, supplied by Millipore, 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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    89
    Millipore goat anti β galactosidase
    Subchronic T 3 treatment enhances the number of <t>β-galactosidase</t> immunopositive cells within the neocortex of Shh +/LacZ mice. Shown are representative images of β-galactosidase expressing cells in the cortex after short-duration T 3 treatment over 2 d in Shh +/LacZ mice. T 3 treatment significantly increased the number of cells that were strongly immunopositive for β-galactosidase in layer V of cortex (A). Results are expressed as a percentage of vehicle-treated control and are the mean ± sem (n = 5/group). *, P
    Goat Anti β Galactosidase, supplied by Millipore, used in various techniques. Bioz Stars score: 89/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Millipore senescence associated β galactosidase sa βgal assay senescence associated β galactosidase
    The treatment with IL6 and IL8 induces senescence in MCF-7 cells. a Representative images of MCF-7 cells treated with SCM during 10 days or ( c ) cytokines (50 ng/ml) during 5 days and stained for <t>SA-β-GAL.</t> Scale bar, 10 μm. b Gene expression profile of p16, p21 and p53 in MCF-7 cells stimulated with SCM or ( e ) cytokines, as indicated. The values were normalized to GADPH and relative to control cells ( dotted lines ). Error bars represent SEM. (* p
    Senescence Associated β Galactosidase Sa βgal Assay Senescence Associated β Galactosidase, supplied by Millipore, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Microglia aged in culture display signs of senescence, including increased senescent-associated β-galactosidase (SA-β-gal) activity and microRNA (miR)-146a expression. Microglial cells were kept in culture for 2 and 16 days in vitro (DIV). Activity of SA-β-gal was determined using a commercial kit. (A) Representative images of 2 and 16 DIV microglia showing SA-β-gal staining. (B) SA-β-gal-positive cells were counted and results expressed in graph bars as mean ± SEM. (C) miR-146a expression was evaluated by Real-Time PCR. Results are expressed in graph bars as mean ± SEM. Cultures, n = 4 per group. t -test, * p

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Microglia change from a reactive to an age-like phenotype with the time in culture

    doi: 10.3389/fncel.2014.00152

    Figure Lengend Snippet: Microglia aged in culture display signs of senescence, including increased senescent-associated β-galactosidase (SA-β-gal) activity and microRNA (miR)-146a expression. Microglial cells were kept in culture for 2 and 16 days in vitro (DIV). Activity of SA-β-gal was determined using a commercial kit. (A) Representative images of 2 and 16 DIV microglia showing SA-β-gal staining. (B) SA-β-gal-positive cells were counted and results expressed in graph bars as mean ± SEM. (C) miR-146a expression was evaluated by Real-Time PCR. Results are expressed in graph bars as mean ± SEM. Cultures, n = 4 per group. t -test, * p

    Article Snippet: Microglial SA-β-gal activity was determined using the Cellular senescence assay kit (Millipore), according to the manufacturer instructions.

    Techniques: Activity Assay, Expressing, In Vitro, Staining, Real-time Polymerase Chain Reaction

    Apoptosis and senescence in tumor sections from control and mixed cells. (A) Sections of tumor tissues were subjected to TUNEL assay. Blue colour shows nuclei of cells stained with DAPI. Green foci show TUNEL positive regions. (B) Tumor sections processed for SA-β-gal assay. The part of image is shown below in the respective inset. Greenish blue color indicates active SA-β-galactosidase.

    Journal: PLoS ONE

    Article Title: Molecular Understanding of Growth Inhibitory Effect from Irradiated to Bystander Tumor Cells in Mouse Fibrosarcoma Tumor Model

    doi: 10.1371/journal.pone.0161662

    Figure Lengend Snippet: Apoptosis and senescence in tumor sections from control and mixed cells. (A) Sections of tumor tissues were subjected to TUNEL assay. Blue colour shows nuclei of cells stained with DAPI. Green foci show TUNEL positive regions. (B) Tumor sections processed for SA-β-gal assay. The part of image is shown below in the respective inset. Greenish blue color indicates active SA-β-galactosidase.

    Article Snippet: Senescence assay Senescence associated β-galactosidase activity was assessed in tumor sections using cellular senescence assay kit (Millipore) as per the instructions provided by the manufacturer.

    Techniques: TUNEL Assay, Staining, β-Gal Assay

    Subchronic T 3 treatment enhances the number of β-galactosidase immunopositive cells within the neocortex of Shh +/LacZ mice. Shown are representative images of β-galactosidase expressing cells in the cortex after short-duration T 3 treatment over 2 d in Shh +/LacZ mice. T 3 treatment significantly increased the number of cells that were strongly immunopositive for β-galactosidase in layer V of cortex (A). Results are expressed as a percentage of vehicle-treated control and are the mean ± sem (n = 5/group). *, P

    Journal: Endocrinology

    Article Title: Thyroid Hormone Regulates the Expression of the Sonic Hedgehog Signaling Pathway in the Embryonic and Adult Mammalian Brain

    doi: 10.1210/en.2010-1396

    Figure Lengend Snippet: Subchronic T 3 treatment enhances the number of β-galactosidase immunopositive cells within the neocortex of Shh +/LacZ mice. Shown are representative images of β-galactosidase expressing cells in the cortex after short-duration T 3 treatment over 2 d in Shh +/LacZ mice. T 3 treatment significantly increased the number of cells that were strongly immunopositive for β-galactosidase in layer V of cortex (A). Results are expressed as a percentage of vehicle-treated control and are the mean ± sem (n = 5/group). *, P

    Article Snippet: In brief, sections were incubated with primary antibody cocktails of goat anti-β-galactosidase with mouse anti-NeuN (1:500; Millipore Corp., Bedford, MA) or mouse anti-RIP (1:10; Developmental Studies Hybridoma Bank, Iowa City, IA) along with rabbit anti-NG2 (1:250; Millipore Corp.) or rabbit anti-GFAP (1:250; Millipore Corp.).

    Techniques: Mouse Assay, Expressing

    The treatment with IL6 and IL8 induces senescence in MCF-7 cells. a Representative images of MCF-7 cells treated with SCM during 10 days or ( c ) cytokines (50 ng/ml) during 5 days and stained for SA-β-GAL. Scale bar, 10 μm. b Gene expression profile of p16, p21 and p53 in MCF-7 cells stimulated with SCM or ( e ) cytokines, as indicated. The values were normalized to GADPH and relative to control cells ( dotted lines ). Error bars represent SEM. (* p

    Journal: Cell Communication and Signaling : CCS

    Article Title: Senescence-associated IL-6 and IL-8 cytokines induce a self- and cross-reinforced senescence/inflammatory milieu strengthening tumorigenic capabilities in the MCF-7 breast cancer cell line

    doi: 10.1186/s12964-017-0172-3

    Figure Lengend Snippet: The treatment with IL6 and IL8 induces senescence in MCF-7 cells. a Representative images of MCF-7 cells treated with SCM during 10 days or ( c ) cytokines (50 ng/ml) during 5 days and stained for SA-β-GAL. Scale bar, 10 μm. b Gene expression profile of p16, p21 and p53 in MCF-7 cells stimulated with SCM or ( e ) cytokines, as indicated. The values were normalized to GADPH and relative to control cells ( dotted lines ). Error bars represent SEM. (* p

    Article Snippet: Senescence-associated β-galactosidase (SA-βGAL) assay Senescence-associated β-galactosidase (SA-βGAL) activity was evaluated in senescent- and young fibroblasts after 2 days of culture by using the cellular senescence assay kit (KAA002, Millipore).

    Techniques: Staining, Expressing