Journal: International Journal of Molecular Sciences

Article Title: Cx43 Promotes Endothelial Cell Migration and Angiogenesis via the Tyrosine Phosphatase SHP-2

doi: 10.3390/ijms23010294

Figure Lengend Snippet: Downregulation of Cx43 reduces endothelial cell migration. ( a ) Western blot analysis of Cx43, Cx40, and Cx37 revealed that Cx43 is expressed in both human umbilical vein endothelial cells (HUVEC) and human microvascular endothelial cells (HMEC), whereas Cx40 and Cx37 are not expressed in HMEC. Cx43 (Cx43), Cx40 (Cx), and Cx37 (Cx) expressing HeLa cells were used as the positive controls. CTL: Wildtype HeLa cell lysate. ( b ) Western blot analysis of Cx43 in HMEC transfected with 100 nM siRNA against Cx43 or non-silencing control siRNA (Ctrl) for 48 h. The Cx43 expression was analysed using a polyclonal Cx43 antibody. Immunostaining for GAPDH was used as the loading control. ( c ) Serum-induced cell migration of HMEC transfected with a Cx43-siRNA or a control siRNA (Ctrl) in response to 10% FCS was assessed in a wound assay. Downregulation of Cx43 significantly decreased cell migration, represented as accumulated distance (*** p < 0.001 Ctrl-siRNA vs. Cx43-siRNA, n = 6, 3 independent cell cultures). ( d ) Representative single cell traces of migrated HMEC.

Article Snippet: For immunoprecipitation, the supernatants were incubated over night at 4 °C with a mouse anti-SHP-2 antibody (Santa Cruz) or a rabbit anti-Cx43 antibody (Sigma Aldrich), and were magnetic beads coated with protein A (Cx43) or G (SHP-2) (Miltenyi Biotec, Bergisch Gladbach, Germany).

Techniques: Migration, Western Blot, Expressing, Transfection, Control, Immunostaining

Journal: International Journal of Molecular Sciences

Article Title: Cx43 Promotes Endothelial Cell Migration and Angiogenesis via the Tyrosine Phosphatase SHP-2

doi: 10.3390/ijms23010294

Figure Lengend Snippet: Downregulation of Cx43 impairs vessel sprouting ex vivo. ( a ) Downregulation of Cx43 in the mouse aortae was achieved by transfection with Cx43 siRNA (500 nM) or control (Ctrl) siRNA, as assessed by Western blot 48 h later. ( b ) Representative images of aortic sprouts in matrigel. Scale bar represents 200 µm. ( c ) The vessel sprouting area (** p < 0.01 Ctrl-siRNA vs. Cx43-siRNA, n = 2 aortae with eight aortic segments each at 3 days; n = 3–4 aortae with 6–8 aortic segments each at 6 days) and ( d ) the number of sprout bifurcations (** p < 0.01 Ctrl-siRNA vs. Cx43-siRNA, n = 2 aortae with 8 aortic segments each at 3 days; *** p < 0.001 Ctrl-siRNA vs. Cx43-siRNA n = 3–4 aortae with 6–8 aortic segments each at 6 days) were significantly reduced upon downregulation of Cx43, as assessed 3 days and 6 days after transfection in a matrigel assay.

Article Snippet: For immunoprecipitation, the supernatants were incubated over night at 4 °C with a mouse anti-SHP-2 antibody (Santa Cruz) or a rabbit anti-Cx43 antibody (Sigma Aldrich), and were magnetic beads coated with protein A (Cx43) or G (SHP-2) (Miltenyi Biotec, Bergisch Gladbach, Germany).

Techniques: Ex Vivo, Transfection, Control, Western Blot, Matrigel Assay

Journal: International Journal of Molecular Sciences

Article Title: Cx43 Promotes Endothelial Cell Migration and Angiogenesis via the Tyrosine Phosphatase SHP-2

doi: 10.3390/ijms23010294

Figure Lengend Snippet: Interaction of Cx43 and SHP-2 induces SHP-2 phosphatase activity. ( a ) Upon immunoprecipitation (IP) of SHP-2 from HeLa cells expressing only Cx43, Cx43 was co-immunoprecipitated. As a control, cells transfected with an empty vector (control (CTL)) were used. L: Whole cell lysates. ( b ) SHP-2 was detected upon immunoblotting after immunoprecipitation of Cx43 from HeLa cells expressing only Cx43 in contrast to the control cells (CTL). L: Whole cell lysates. Images originates from the same blot, which was cropped; ( c ) SHP-2 was immunoprecipitated together with Cx43 in HMEC; ( d ) Co-localization of SHP-2 and Cx43 was detected by immunofluorescent staining of HMEC followed by confocal microscopic imaging. Scale bar represents 20 µm; ( e ) SHP-2 phosphatase activity was significantly increased in HeLa cells expressing Cx43 compared to HeLa cells transfected with an empty vector (* p < 0.05, n = 4 independent cell cultures), as assessed by detection of dephosphorylation of a phosphate analogue in SHP-2 immunoprecipitates.

Article Snippet: For immunoprecipitation, the supernatants were incubated over night at 4 °C with a mouse anti-SHP-2 antibody (Santa Cruz) or a rabbit anti-Cx43 antibody (Sigma Aldrich), and were magnetic beads coated with protein A (Cx43) or G (SHP-2) (Miltenyi Biotec, Bergisch Gladbach, Germany).

Techniques: Activity Assay, Immunoprecipitation, Expressing, Control, Transfection, Plasmid Preparation, Western Blot, Staining, Imaging, De-Phosphorylation Assay

Journal: International Journal of Molecular Sciences

Article Title: Cx43 Promotes Endothelial Cell Migration and Angiogenesis via the Tyrosine Phosphatase SHP-2

doi: 10.3390/ijms23010294

Figure Lengend Snippet: Inactivation of SHP-2 reduces endothelial cell migration. ( a ) Overexpression of a dominant negative substrate trapping mutant SHP-2 (SHP-2 CS) in HMEC significantly reduced serum (10%) induced endothelial cell migration, as assessed by the accumulated distance in µm in an in vitro wound assay (* p < 0.05, n = 3 independent cell cultures) compared to SHP-2 WT. Additional knock-down of Cx43 (Cx43 siRNA) did not further decrease migration (ns: not significant, n = 3 independent cell cultures). ( b ) Representative single cell traces of migrated HMEC.

Article Snippet: For immunoprecipitation, the supernatants were incubated over night at 4 °C with a mouse anti-SHP-2 antibody (Santa Cruz) or a rabbit anti-Cx43 antibody (Sigma Aldrich), and were magnetic beads coated with protein A (Cx43) or G (SHP-2) (Miltenyi Biotec, Bergisch Gladbach, Germany).

Techniques: Migration, Over Expression, Dominant Negative Mutation, Mutagenesis, In Vitro, Knockdown

Journal: International Journal of Molecular Sciences

Article Title: Cx43 Promotes Endothelial Cell Migration and Angiogenesis via the Tyrosine Phosphatase SHP-2

doi: 10.3390/ijms23010294

Figure Lengend Snippet: The Cx43 and SHP-2 interaction is vital for endothelial migration. ( a ) Treatment of HMEC overexpressing a constitutively active mutant SHP-2 (SHP-2 EA) with Cx43 siRNA did not rescue the reduced migratory response caused by Cx43 knock-down (Cx43 siRNA) (ns: not significant, n = 3 independent cell cultures) compared to SHP-2 EA cells treated with control siRNA (Ctrl) (* p < 0.05, n = 3 independent cell cultures), as assessed by the accumulated distance in µm in an in vitro wound assay (* p < 0.05, n = 3 independent cell cultures). ( b ) Representative single cell traces of migrated HMEC.

Article Snippet: For immunoprecipitation, the supernatants were incubated over night at 4 °C with a mouse anti-SHP-2 antibody (Santa Cruz) or a rabbit anti-Cx43 antibody (Sigma Aldrich), and were magnetic beads coated with protein A (Cx43) or G (SHP-2) (Miltenyi Biotec, Bergisch Gladbach, Germany).

Techniques: Migration, Mutagenesis, Knockdown, Control, In Vitro

Journal: Korean Circulation Journal

Article Title: Preventive Effects of the Angiotensin-II Receptor Blocker on Atrial Remodeling in an Ischemic Heart Failure Model of Rats

doi: 10.4070/kcj.2013.43.10.686

Figure Lengend Snippet: Even distribution of connexin 43 protein on the intercalated disc was observed in the sham group (A: magnification: ×400). Connexin 43 protein stain was not observed on the intercalated disc in the heart failure group (B: magnification: ×400). A slight connexin 43 protein stain was observed on the intercalated disc in the heart failure-ARB group (C: magnification: ×400). Western blot of connexin 43 protein showed a lower expression in the heart failure-ARB group (D). ARB: angiotensin-II receptor blocker.

Article Snippet: The immunohistochemical stain for connexin 43 (rabbit polyclonal anti-Cx43 antibody, 1/100, Zymed-laboratories, CliniSciences, France) showed a significant amount of plasmalemmal distribution of connexin 43 in the heart failure group.

Techniques: Staining, Western Blot, Expressing

Journal: Nature Communications

Article Title: Physiological synaptic activity and recognition memory require astroglial glutamine

doi: 10.1038/s41467-022-28331-7

Figure Lengend Snippet: a Representative confocal images showing close proximity of Cx43 (cyan) in GFAP-eGFP positive astrocytes (yellow) to presynaptic structures immunolabeled for VGlut1 (magenta). Higher magnification images of a region containing astroglial processes (blue square) are shown in the middle row. Arrowheads denote points of close contact. Masks showing co-localized area of GFAP (yellow) and Cx43 (cyan) as total Cx43 (binary inverse image) and co-localized area of total Cx43 (cyan) and VGlut1(magenta) as presynaptic Cx43 (binary inverse image). b Bar graph (mean ± SEM) showing % Perisynaptic Cx43 normalized to total Cx43 area ( n = 13 fields, 3 independent experiments). c Schematic illustration of co-purification of perisynaptic astroglial processes in crude synaptosomes. d Representative western blots showing an enrichment of Cx43 protein in synaptosomal preparations (Syn) compared to total hippocampal lysates (Hip) in wild type (+/+), but not in glial conditional Cx43 knockout (−/−) mice. GAPDH was used as a loading control. e Representative high magnification electron micrographs showing the presence of Cx43 protein labeled by immunogold particles in astroglial processes near synaptic complexes. f Distribution histogram of distance between Cx43 gold grains and the nearest active zone. Scale bars: a 5 µm, e 0.5 µm (left), 0.3 µm (right). Representative images ( a ) are from replicates described in ( b ); d are from n = 3 replicates; and e are from n = 8 replicates. Source data are provided as a Source data file.

Article Snippet: The following primary antibodies were used: polyclonal chicken anti-GFP (1:500, AB13970, Abcam), monoclonal mouse anti-VGlut1 (1:200, 135511, Synaptic Systems), monoclonal mouse anti-Cx43 (1:500, 610-062, BD Biosciences), polyclonal rabbit anti-Cx43 (1:500, 71-2200, Zymed Laboratories), monoclonal mouse anti-GFAP (1:500, G3893, Sigma), and monoclonal mouse anti-GAPDH-peroxidase (1:1000, G9295, Sigma).

Techniques: Immunolabeling, Copurification, Western Blot, Knock-Out, Labeling

Journal: Nature Communications

Article Title: Physiological synaptic activity and recognition memory require astroglial glutamine

doi: 10.1038/s41467-022-28331-7

Figure Lengend Snippet: Acute hippocampal slices were loaded with ethidium bromide (EtBr) under different experimental conditions. a Sample images of EtBr uptake (magenta) in hippocampal GFAP-immunolabeled astrocytes (green) are shown for Control, Stim (10 Hz, 30 s every 3 min for 20 min) in the absence or presence of the Cx43 HC blocker Gap26 or a Gap26 scramble version (Src). Higher magnifications of the CA1 stratum radiatum subregion are shown in bottom two rows. b Schematic illustrating stimulation of hippocampal Schaffer collaterals and EtBr uptake in neighboring astrocytes. c Quantification of EtBr uptake normalized to 100% control (dotted line) is shown. Stimulation-enhanced EtBr uptake by nearly 2-fold (mean ± SEM; Control, n = 6; Stim, n = 7, p = 0.0002 between Control and Stim, one-sampled t -test). This enhanced uptake was not observed in the presence of Gap26 (Stim + Gap26, n = 4, p < 0.0001 between Stim and Stim+Gap26, one-way ANOVA with Bonferroni’s post hoc test; p = 0.0168 with control, one-sampled t -test) but persisted with Src (Stim + Src, n = 4, p = 0.6857 between Stim and Stim+Src, one-way ANOVA with Bonferroni’s post hoc test; p = 0.0125 with control, one-sampled t -test), while Gap26 alone decreases EtBr uptake from control level ( n = 5, p = 0.014, one-sampled t -test) but not Src ( n = 5, p = 0.4443, one-sampled t -test). Scale bars: a 450 µm (top), 50 µm (middle and bottom). Asterisks indicate statistical significance (*** p < 0.001; * p < 0.05). Source data are provided as a Source data file.

Article Snippet: The following primary antibodies were used: polyclonal chicken anti-GFP (1:500, AB13970, Abcam), monoclonal mouse anti-VGlut1 (1:200, 135511, Synaptic Systems), monoclonal mouse anti-Cx43 (1:500, 610-062, BD Biosciences), polyclonal rabbit anti-Cx43 (1:500, 71-2200, Zymed Laboratories), monoclonal mouse anti-GFAP (1:500, G3893, Sigma), and monoclonal mouse anti-GAPDH-peroxidase (1:1000, G9295, Sigma).

Techniques: Immunolabeling

Journal: Nature Communications

Article Title: Physiological synaptic activity and recognition memory require astroglial glutamine

doi: 10.1038/s41467-022-28331-7

Figure Lengend Snippet: Acute hippocampal slices were loaded with ethidium bromide (EtBr) under stimulated conditions in the absence or presence of various blockers. a , b Sample images of EtBr uptake (magenta) in hippocampal GFAP-immunolabeled astrocytes (green) are shown in ( a ) and quantified in ( b ). Stimulation-enhanced uptake was blocked in the presence of NBQX + CPP (20 μM, n = 6, p = 0.0112 with wild-type control, +/+, n = 11, one-way ANOVA with Bonferroni’s post hoc test), but not LY341495 (20 μM; n = 5, p = 0.2433 with +/+, one-way ANOVA with Bonferroni’s post hoc test), indicating the specific involvement of ionotropic glutamate receptor activity. This stimulation-dependent uptake was also blocked in the presence of BaCl 2 (200 μM; n = 7, p = 0.0004 with +/+, one-way ANOVA with Bonferroni’s post hoc test) and in acute slices prepared from Kir4.1−/− mice ( n = 5, p = 0.0086 with +/+, one-way ANOVA with Bonferroni’s post hoc test), indicating the specific involvement of Kir4.1 activity. c , d Incubation of acute slices with either 2 mM K + or 1 μM glutamate (Glut) alone enhanced basal EtBr uptake (K + + Src, n = 9, 3 experiments, p = 0.0061; Glut+Src, n = 9, 3 experiments, p = 0.0056 with Src, n = 14, 5 experiments; Kruskal–Wallis test with Dunn’s post hoc test) in a Cx43 HC-dependent manner (K + + Gap26, n = 20, 7 experiments, p < 0.0001 with K + + Src; Glut + Gap26, n = 11, 5 experiments, p < 0.0001 with Glut+Src; Kruskal–Wallis test with Dunn’s post hoc test). In Kir4.1−/− hippocampal slices, neither K + nor Glut were able to enhance EtBr uptake (K + Kir4.1−/−, n = 49, 17 experiments, p < 0.0001 with K + +/+, n = 22, 8 experiments; Glut Kir4.1−/−, n = 27, 9 experiments, p = 0.0001 with Glut +/+, n = 23, 8 ex p eriments, Mann–Whitney test). Mean ± SEM in ( b ) and ( d ). Scale bars: a and c , 50 µm. Asterisks indicate statistical significance (*** p < 0.001; ** p < 0.01; * p < 0.05). NBQX = 2,3-Dihydroxy-6-nitro-7-sulfamoyl-benzo[f]chinoxalin-2,3-dion; CPP = (3-((R)-2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid. Source data are provided as a Source data file.

Article Snippet: The following primary antibodies were used: polyclonal chicken anti-GFP (1:500, AB13970, Abcam), monoclonal mouse anti-VGlut1 (1:200, 135511, Synaptic Systems), monoclonal mouse anti-Cx43 (1:500, 610-062, BD Biosciences), polyclonal rabbit anti-Cx43 (1:500, 71-2200, Zymed Laboratories), monoclonal mouse anti-GFAP (1:500, G3893, Sigma), and monoclonal mouse anti-GAPDH-peroxidase (1:1000, G9295, Sigma).

Techniques: Immunolabeling, Activity Assay, Incubation, MANN-WHITNEY

Journal: Nature Communications

Article Title: Physiological synaptic activity and recognition memory require astroglial glutamine

doi: 10.1038/s41467-022-28331-7

Figure Lengend Snippet: a Representative confocal (dark background) and thresholded binary (white background) images of hippocampal CA1 astrocytes dialyzed with RhGln (0.8 mM, 20 min) via the patch pipette under control or stimulated (10 Hz, 30 s every 3 min for 20 min) conditions in acute slices obtained from: wild-type mice (+/+), glial conditional Cx43 knockout mice (−/−) or wild-type mice exposed to Gap26 (+/+ Gap26) or Gap26 scramble (+/+Src) peptides. The binary images were quantified by Sholl analysis ( b ) and total punctate area ( c ). In +/+ mice, repetitive synaptic stimulation strongly increased the punctate RhGln-labeling compared to control as shown by both Sholl analysis (+/+: Ct, n = 12; Stim, n = 9, p < 0.0001, two-way ANOVA in b ) and total punctate area ( p < 0.0001 between Ct and Stim in +/+, one-way ANOVA with Bonferroni’s post hoc test in c ). This was abolished in −/− mice (−/−: Ct, n = 7; Stim, n = 5, p = 0.0826, two-way ANOVA in b ; p > 0.999 between Ct and Stim in −/−, and p = 0.0002 between +/+ Stim and −/− Stim, one-way ANOVA with Bonferroni’s post hoc test in c ) or in the presence of Gap26 (+/+ Gap26: Ct, n = 8; Stim, n = 5, p = 0.0651, two-way ANOVA in b ; p > 0.999 between Ct and Stim in +/+ Gap26, and p < 0.0001 between Stim of +/+ and +/+ Gap26, one-way ANOVA with Bonferroni’s post hoc test in c ), but unchanged in the presence of the scramble Gap26 peptide (+/+Src: Ct, n = 5; Stim, n = 4, p < 0.0001, two-way ANOVA in b ; p = 0.025 between Ct and Stim with +/+ Src, and p < 0.0001 between Stim of +/+ Gap26 and +/+ Src, one-way ANOVA with Bonferroni’s post hoc test in c ). Gap26 alone also inhibited the spread of glutamine, suggesting a basal transfer of glutamine into synaptic structures which is dependent on Cx43 HC activity ( p < 0.0001 between +/+ Gap26 Ct and +/+ Gap26 Stim, two-way ANOVA in b ). Mean ± SEM in ( b ), ( c ). Scale bar: a 20 µm. Asterisks indicate statistical significance (*** p < 0.001; * p < 0.05). Source data are provided as a Source data file.

Article Snippet: The following primary antibodies were used: polyclonal chicken anti-GFP (1:500, AB13970, Abcam), monoclonal mouse anti-VGlut1 (1:200, 135511, Synaptic Systems), monoclonal mouse anti-Cx43 (1:500, 610-062, BD Biosciences), polyclonal rabbit anti-Cx43 (1:500, 71-2200, Zymed Laboratories), monoclonal mouse anti-GFAP (1:500, G3893, Sigma), and monoclonal mouse anti-GAPDH-peroxidase (1:1000, G9295, Sigma).

Techniques: Transferring, Knock-Out, Labeling, Activity Assay

Journal: Nature Communications

Article Title: Physiological synaptic activity and recognition memory require astroglial glutamine

doi: 10.1038/s41467-022-28331-7

Figure Lengend Snippet: a Sample image of the hippocampus of Cx43−/− mice injected intra-hippocampally with rAAV2/9-GFAP-Cx43-GFP virus, showing numerous cells expressing Cx43-GFP in the CA1 area. The blue box is magnified on the right. b RhGln (magenta) was loaded into a Cx43-GFP expressing astrocyte (yellow) via a patch pipette as shown. Arrow head indicates the patched cell. c Sample images after immunostaining showing specific expression of Cx43 (cyan) in Cx43-GFP-positive (yellow) astrocytes (GFAP, magenta). The yellow box is magnified in the bottom row. Solid and dotted white lines outline GFP-positive and -negative astrocytes, respectively. d – g Cx43−/− mice first received either rAAV2/9-GFAP-Cx43-GFP (−/− Cx43 Rescue, d left, e and g ) or rAAV2/9-GFAP-GFP (−/− GFP Control, d right, f and g ) virus. Hippocampal astrocytes were then dialyzed with RhGln under either control or synaptic stimulation (10 Hz, 30 s) conditions for 20 min. Both representative confocal (dark background) and thresholded binary (white background) images are shown in ( d ) for each condition. The binary images were quantified by Sholl analysis ( e , f ) and total punctate area ( g ). The stimulation-induced transfer of RhGln was rescued in Cx43−/− mice ( n = 5) by restoring Cx43 expression selectively in astrocytes via viral infection shown by both Sholl analysis (−/− Cx43 rescue, n = 4, p < 0.0001, two-way ANOVA for e ) and total punctate area ( p = 0.0031 between Ct and Stim with −/− Cx43 Rescue, one-way ANOVA with Bonferroni’s post hoc test for g ) as compared to the GFP control infection (−/− GFP Ct, n = 3; Stim, n = 3, p = 0.9856, two-way ANOVA for f , p > 0.999 between Ct and Stim with −/−GFP Control and p < 0.0001 between Stim of −/− Cx43 Rescue and −/− GFP Control, one-way ANOVA with Bonferroni’s post hoc test for g ). Mean ± SEM in ( e )–( g ). Scale bars: a 200 µm (left) and 50 µm (right); b 50 µm (top) and 20 µm (bottom); c , d 20 µm. Asterisks indicate statistical significance (*** p < 0.001; ** p < 0.01). Representative images a , b from replicates described in ( g ); c are from n = 3 replicates. Source data are provided as a Source data file.

Article Snippet: The following primary antibodies were used: polyclonal chicken anti-GFP (1:500, AB13970, Abcam), monoclonal mouse anti-VGlut1 (1:200, 135511, Synaptic Systems), monoclonal mouse anti-Cx43 (1:500, 610-062, BD Biosciences), polyclonal rabbit anti-Cx43 (1:500, 71-2200, Zymed Laboratories), monoclonal mouse anti-GFAP (1:500, G3893, Sigma), and monoclonal mouse anti-GAPDH-peroxidase (1:1000, G9295, Sigma).

Techniques: Injection, Expressing, Transferring, Immunostaining, Infection

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: The Dome Wall of Bladder Acts as a Pacemaker Site in Detrusor Instability in Rats

doi: 10.12659/MSM.904406

Figure Lengend Snippet: List of primers used in PCR.

Article Snippet: Tissues sections were incubated with primary antibody dilutions made up in 0.2% BSA/0.01 MPBS: goat anti-mouse c-KIT (1: 100, Santa Cruz, Germany), rabbit anti-mouse HCN2 (1: 100, Chemicon, Japan), and rabbit anti-mouse Cx43 (1: 100, Cell Signaling, USA) overnight at 4°C, respectively.

Techniques: Sequencing

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: The Dome Wall of Bladder Acts as a Pacemaker Site in Detrusor Instability in Rats

doi: 10.12659/MSM.904406

Figure Lengend Snippet: The expression of Connexin43. ( A ) Connexin43 mRNAs and ( B ) protein expression in DI group. ( C ) Co-expression of Connexin-43 and c-kit-positive ICCs in the DI detrusor. * P <0.05 vs. other parts in DI group.

Article Snippet: Tissues sections were incubated with primary antibody dilutions made up in 0.2% BSA/0.01 MPBS: goat anti-mouse c-KIT (1: 100, Santa Cruz, Germany), rabbit anti-mouse HCN2 (1: 100, Chemicon, Japan), and rabbit anti-mouse Cx43 (1: 100, Cell Signaling, USA) overnight at 4°C, respectively.

Techniques: Expressing

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: The Dome Wall of Bladder Acts as a Pacemaker Site in Detrusor Instability in Rats

doi: 10.12659/MSM.904406

Figure Lengend Snippet: The expression of Connexin43. ( A ) HCN2 mRNAs and ( B ) protein expression in DI group. ( C ) Co-expression of HCN2 and c-kit-positive ICCs in the DI detrusor. * P <0.05 vs. other parts in DI group.

Article Snippet: Tissues sections were incubated with primary antibody dilutions made up in 0.2% BSA/0.01 MPBS: goat anti-mouse c-KIT (1: 100, Santa Cruz, Germany), rabbit anti-mouse HCN2 (1: 100, Chemicon, Japan), and rabbit anti-mouse Cx43 (1: 100, Cell Signaling, USA) overnight at 4°C, respectively.

Techniques: Expressing