rabbit anti cx43  (Proteintech)

Journal: IBRO neuroscience reports

Article Title: Acupuncture regulates astrocyte neurotoxic polarization to protect blood-brain barrier integrity in delayed thrombolysis through mediating ERK1/2/Cx43 axis.

doi: 10.1016/j.ibneur.2025.04.005

Figure Lengend Snippet: Fig. 2. Transcriptomic analysis reveals dynamic regulation of astrocytes neurotoxic polarization by accupuncture intervention. (A) Principal component analysis showing the separation between groups. The barplots in the right corner displayed the number of differentially expressed genes (DEGs) between the Acu + 6 h rt-PA vs 6 h rt-PA groups. (B) GO enrichment analysis highlighting significant astrocyte-related pathways in the Acu + 6 h rt-PA vs 6 h rt-PA comparison. (C) Electron microscopy images of astrocyte endfeet (Scale bar=2μm). (D) t-SNE clustering of single-cell data from Sham and MCAO/R groups, with 13 distinct cell types an notated. (E) tSNE plot showing higher expression of astrocyte active marker in the MCAO/R groups (F) Dotplot showing the expression of GFAP and C3. (G) Neurotoxic polarization activity scores in astrocytes from the Sham and MCAO/R groups. (H) Differential gene expression in astrocytes showing up-regulation of inflammatory and chemokine genes (Ccl4, Ccl12, Cxcl2, Ccl3, Tnfrsf1a, SerpinA3N) and the gap junction gene Cx43 in the MCAO/R group. (I) Venn diagram showing the intersection of DEGs in astrocytes of MCAO/R vs. Sham and that of Acu + 6 h rt-PA vs 6 h rt-PA groups. (J) Left: the average change in the expression of DEGs in (H). Right: The heatmap showing the expression of represent genes related to inflammatory and chemokine. Genes highlighted in red are the genes of interest. Data was presented as mean ± SD, *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: Membranes were incubated overnight at 4◦C with primary antibodies: C3 (1:1000, 21337–1-AP, Proteintech, China), GFAP (1:2000, 3670, CST, USA), Cx43 (1:1000, 83649S, CST, China), Phospho-Connexin 43 (Ser368) (1:1000, 3511, CST, USA), ERK1/2 (1:1000, 4695, CST, USA), Phospho-ERK1/2(Thr202/Tyr204) (1:2000, 4370, CST, USA), ZO-1 (1:1000, 21773–1-AP, Proteintech, China), β-actin (1:100000, AC026, Abclonal, China), β-tublin (1:5000, 30302ES20, Yeasen, China)and GAPDH (1:200000, 60004–1-Ig, Proteintech, China).

Techniques: Comparison, Electron Microscopy, Expressing, Marker, Activity Assay, Gene Expression

Journal: IBRO neuroscience reports

Article Title: Acupuncture regulates astrocyte neurotoxic polarization to protect blood-brain barrier integrity in delayed thrombolysis through mediating ERK1/2/Cx43 axis.

doi: 10.1016/j.ibneur.2025.04.005

Figure Lengend Snippet: Fig. 4. Cx43 inhibition reduces neurotoxic polarization of astrocytes and inflammatory responses. (A) Representative IF images showed co-localization of Cx43 (red) and GFAP (Green) in the Sham, rt-PA, and Tat-Gap19-treated groups. (B) Quantification of GFAP+ /Cx43+ intensities in the brain cortex of ischemic penumbra from different groups (n = 3, Scale bar=20μm). (C) Western blot analysis of C3 expression in the groups. (D) Quantification of C3 protein expression in the western blot analysis (n = 3). (E) Double immunofluorescence staining for C3 (red) and GFAP (green) in the ischemic penumbra. (F) the number of C3+/GFAP+ cells were quantified in each 1 mm2 area of the ischemic penumbra (n = 3, Scale bar=20μm). (G-I) Secretion of inflammatory cytokines IL-1β, IL-6 and TNF-α (n = 5). Data was presented as mean ± SD, *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: Membranes were incubated overnight at 4◦C with primary antibodies: C3 (1:1000, 21337–1-AP, Proteintech, China), GFAP (1:2000, 3670, CST, USA), Cx43 (1:1000, 83649S, CST, China), Phospho-Connexin 43 (Ser368) (1:1000, 3511, CST, USA), ERK1/2 (1:1000, 4695, CST, USA), Phospho-ERK1/2(Thr202/Tyr204) (1:2000, 4370, CST, USA), ZO-1 (1:1000, 21773–1-AP, Proteintech, China), β-actin (1:100000, AC026, Abclonal, China), β-tublin (1:5000, 30302ES20, Yeasen, China)and GAPDH (1:200000, 60004–1-Ig, Proteintech, China).

Techniques: Inhibition, Western Blot, Expressing, Double Immunofluorescence Staining

Journal: IBRO neuroscience reports

Article Title: Acupuncture regulates astrocyte neurotoxic polarization to protect blood-brain barrier integrity in delayed thrombolysis through mediating ERK1/2/Cx43 axis.

doi: 10.1016/j.ibneur.2025.04.005

Figure Lengend Snippet: Fig. 5. Acupuncture inhibits Cx43 expression in neurotoxic astrocytes. (A) Western blot analysis of Cx43 and p-Cx43 expression in the Sham, model, rt-PA, and rt- PA+Acu groups. (B,C) Quantification of Cx43 and p-Cx43 protein expression in the western blot analysis (n = 3). (D) Correlation analysis between Cx43 and C3 expression, and p-Cx43 and C3 expression. (E) Double immunofluorescence staining for Cx43 (red) and GFAP (green) in the ischemic penumbra (Scale bar=20μm). Data was presented as mean ± SD, **p < 0.01, ***p < 0.001.

Article Snippet: Membranes were incubated overnight at 4◦C with primary antibodies: C3 (1:1000, 21337–1-AP, Proteintech, China), GFAP (1:2000, 3670, CST, USA), Cx43 (1:1000, 83649S, CST, China), Phospho-Connexin 43 (Ser368) (1:1000, 3511, CST, USA), ERK1/2 (1:1000, 4695, CST, USA), Phospho-ERK1/2(Thr202/Tyr204) (1:2000, 4370, CST, USA), ZO-1 (1:1000, 21773–1-AP, Proteintech, China), β-actin (1:100000, AC026, Abclonal, China), β-tublin (1:5000, 30302ES20, Yeasen, China)and GAPDH (1:200000, 60004–1-Ig, Proteintech, China).

Techniques: Expressing, Western Blot, Double Immunofluorescence Staining

Journal: IBRO neuroscience reports

Article Title: Acupuncture regulates astrocyte neurotoxic polarization to protect blood-brain barrier integrity in delayed thrombolysis through mediating ERK1/2/Cx43 axis.

doi: 10.1016/j.ibneur.2025.04.005

Figure Lengend Snippet: Fig. 6. Acupuncture modulates the polarization-related ERK1/2-Cx43 pathway of astrocyte. (A) Gene set enrichment analysis (GSEA) showing significant enrich ment of the ERK1/2 pathway in neurotoxic polarized astrocytes. (B) GSEA showing downregulation of the ERK1/2 pathway after acupuncture intervention in delayed reperfusion. (C) Western blot analysis of p-ERK1/2 and p-Cx43 expression in the rt-PA and C6 + Acu groups. (D, E) Quantification of p-ERK1/2 and p-Cx43 protein expression from western blot analysis (n = 3). (F) Double immunofluorescence staining for p-ERK1/2 (red) and GFAP (green) in astrocytes. (G) Quantification of the intensity of pERK+/GFAP+ cells in each 1 mm2 area of the ischemic penumbra (n = 3, Scale bar=20μm). Data was presented as mean ± SD, *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: Membranes were incubated overnight at 4◦C with primary antibodies: C3 (1:1000, 21337–1-AP, Proteintech, China), GFAP (1:2000, 3670, CST, USA), Cx43 (1:1000, 83649S, CST, China), Phospho-Connexin 43 (Ser368) (1:1000, 3511, CST, USA), ERK1/2 (1:1000, 4695, CST, USA), Phospho-ERK1/2(Thr202/Tyr204) (1:2000, 4370, CST, USA), ZO-1 (1:1000, 21773–1-AP, Proteintech, China), β-actin (1:100000, AC026, Abclonal, China), β-tublin (1:5000, 30302ES20, Yeasen, China)and GAPDH (1:200000, 60004–1-Ig, Proteintech, China).

Techniques: Western Blot, Expressing, Double Immunofluorescence Staining

Journal: IBRO neuroscience reports

Article Title: Acupuncture regulates astrocyte neurotoxic polarization to protect blood-brain barrier integrity in delayed thrombolysis through mediating ERK1/2/Cx43 axis.

doi: 10.1016/j.ibneur.2025.04.005

Figure Lengend Snippet: Fig. 7. Acupuncture suppresses neurotoxic polarization of astrocytes via the ERK1/2-Cx43 pathway. (a) Western blot analysis of C3 expression. (B) Quantification of p-ERK1/2 and p-Cx43 protein expression from western blot analysis (n = 3). (C) Double immunofluorescence staining for Cx43 (red) and GFAP (green). (D) Quantification of C3+/GFAP+ cells in the different groups (n = 3, Scale bar=20μm). Data was presented as mean ± SD, *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: Membranes were incubated overnight at 4◦C with primary antibodies: C3 (1:1000, 21337–1-AP, Proteintech, China), GFAP (1:2000, 3670, CST, USA), Cx43 (1:1000, 83649S, CST, China), Phospho-Connexin 43 (Ser368) (1:1000, 3511, CST, USA), ERK1/2 (1:1000, 4695, CST, USA), Phospho-ERK1/2(Thr202/Tyr204) (1:2000, 4370, CST, USA), ZO-1 (1:1000, 21773–1-AP, Proteintech, China), β-actin (1:100000, AC026, Abclonal, China), β-tublin (1:5000, 30302ES20, Yeasen, China)and GAPDH (1:200000, 60004–1-Ig, Proteintech, China).

Techniques: Western Blot, Expressing, Double Immunofluorescence Staining

Journal: IBRO neuroscience reports

Article Title: Acupuncture regulates astrocyte neurotoxic polarization to protect blood-brain barrier integrity in delayed thrombolysis through mediating ERK1/2/Cx43 axis.

doi: 10.1016/j.ibneur.2025.04.005

Figure Lengend Snippet: Fig. 8. Acupuncture attenuates neuroinflammation and promotes blood-brain barrier recovery via the ERK1/2/Cx43 pathway. (A-C) Concentrations of inflammatory factors IL-1β, IL-6 and TNF-α were measured across experimental groups (n = 6). (D) Representative images of BBB leakage assessed by EB fluorescence. (E) Immunofluorescence staining of tight junction protein (ZO-1, green) and endothelial cell (CD31, red) shows blood-brain barrier recovery. (F) Western blot analysis of ZO-1 expression in different groups. (G) Quantification of ZO-1 protein expression from Western blot analysis (n = 3). Data was presented as mean ± SD, *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: Membranes were incubated overnight at 4◦C with primary antibodies: C3 (1:1000, 21337–1-AP, Proteintech, China), GFAP (1:2000, 3670, CST, USA), Cx43 (1:1000, 83649S, CST, China), Phospho-Connexin 43 (Ser368) (1:1000, 3511, CST, USA), ERK1/2 (1:1000, 4695, CST, USA), Phospho-ERK1/2(Thr202/Tyr204) (1:2000, 4370, CST, USA), ZO-1 (1:1000, 21773–1-AP, Proteintech, China), β-actin (1:100000, AC026, Abclonal, China), β-tublin (1:5000, 30302ES20, Yeasen, China)and GAPDH (1:200000, 60004–1-Ig, Proteintech, China).

Techniques: Fluorescence, Immunofluorescence, Staining, Western Blot, Expressing

Journal: Cell Death & Disease

Article Title: Cytoplasmic connexin43-microtubule interactions promote glioblastoma stem-like cell maintenance and tumorigenicity

doi: 10.1038/s41419-025-07514-2

Figure Lengend Snippet: A VTC-001, VTC-034, and VTC-037 GSCs were cultured in stem cell medium as gliospheres (GSC-sph) or as adherent cells (GSC-adh), or differentiated for 1 to 3 days in medium containing 10% FBS (GSC-diff), and observed by phase-contrast microscopy. Scale bar: 200 μm. B VTC-001, VTC-034, and VTC-037 cultured as GSC-derived gliospheres (sph), adherent GSCs (adh), or differentiated cells after addition of 10% FBS for 1, 2, 3, and 7 days, were lysed and analyzed by immunoblotting using antibodies against CD133, Olig2, and GAPDH as loading control. C VTC-001, VTC-034, and VTC-037 GSCs were differentiated (Diff) or not for 24 h, and cell lysates were analyzed by immunoblotting for SOX2, and GAPDH as loading control (quantification shown on the right; n = 3). A two-tailed unpaired Student’s t -test was used; * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001. Data are represented as mean ± SD. D VTC-001, VTC-034, and VTC-037 GSCs were differentiated (Diff) or not for 24 h, and cell lysates were analyzed by immunoblotting for Notch1, and GAPDH as loading control. E VTC-001, VTC-034, and VTC-037 GSCs were differentiated (Diff) or not for 24 h, and cell lysates were analyzed by immunoblotting for Cx43, and GAPDH as loading control.

Article Snippet: Primary antibodies rabbit anti-Cx43 (MilliporeSigma; C6219), mouse anti-pan Cadherin (MilliporeSigma; C1821), or mouse anti-α-tubulin (MilliporeSigma; T6199) were diluted in blocking buffer and labeling was performed overnight at 4 °C.

Techniques: Cell Culture, Microscopy, Derivative Assay, Western Blot, Control, Two Tailed Test

Journal: Cell Death & Disease

Article Title: Cytoplasmic connexin43-microtubule interactions promote glioblastoma stem-like cell maintenance and tumorigenicity

doi: 10.1038/s41419-025-07514-2

Figure Lengend Snippet: VTC-001, VTC-034, and VTC-037 GSCs ( A ), or normal human astrocytes (NHA; B ) in culture were fixed and immunostained using antibodies against Cx43 (green), and pan-Cadherin (pan-Cad – red) to stain cell borders. Confocal immunofluorescence was used to observe Cx43 subcellular localization in the cytoplasm and at the cell borders. DAPI was used to stain nuclei. Scale bar: 20 μm. C VTC-037 GSC protein lysates were subjected to fractionation using a Triton X-100 solubility assay, and analyzed by immunoblotting for total, soluble (non-junctional), and insoluble (junctional) Cx43. Quantification of insoluble (junctional) Cx43 band intensity relative to soluble (non-junctional) shown on the right ( n = 3). A two-tailed unpaired Student’s t -test was used; ** p ≤ 0.01. Data are represented as mean ± SD. D Stochastic optical reconstruction microscopy (STORM) derived localizations of Cx43 (green) and α-tubulin (magenta) in VTC-001 GSCs or differentiated by addition of 10% FBS for 24 h. Left 6 panels: point-splatting visualization of STORM localizations to better identify complexing (white; scale bar: 5 μm). Right 4 panels: point-clouds of 50 nm spheres representing individual localizations, including zoomed-in regions (scale bar: 1 μm). E Cross-pair correlation functions for Cx43/α-tubulin complexing in VTC-001, VTC-034, and VTC-037 GSCs versus differentiated cell populations. ( n = 10). F VTC-001 and VTC-037 GSCs were differentiated or not, and cell lysates were subjected to co-immunoprecipitation using Cx43 antibody or IgG for negative control, and/or immunoblotted using antibodies against Cx43, β-tubulin, and GAPDH for loading control. Black arrow indicates β-tubulin band. G Densitometry quantification of β-tubulin levels co-immunoprecipitating with Cx43 in differentiated cells relative to GSC counterparts ( n = 3; decreased: 40.3%, 21.1%, and 45.9% in differentiated VTC-001; decreased: 51.4%, 65.6%, and 75.5% in differentiated VTC-037). A two-tailed unpaired Student’s t -test was used; * p ≤ 0.05.

Article Snippet: Primary antibodies rabbit anti-Cx43 (MilliporeSigma; C6219), mouse anti-pan Cadherin (MilliporeSigma; C1821), or mouse anti-α-tubulin (MilliporeSigma; T6199) were diluted in blocking buffer and labeling was performed overnight at 4 °C.

Techniques: Staining, Immunofluorescence, Fractionation, Solubility, Western Blot, Two Tailed Test, Microscopy, Derivative Assay, Immunoprecipitation, Negative Control, Control

Journal: Cell Death & Disease

Article Title: Cytoplasmic connexin43-microtubule interactions promote glioblastoma stem-like cell maintenance and tumorigenicity

doi: 10.1038/s41419-025-07514-2

Figure Lengend Snippet: A Schematic of the JM2 peptide that encompasses the Cx43 tubulin-binding domain, an antennapedia cell penetration domain, and a biotin tag for tracking. Control peptides include JM2-scrambled and antennapedia. B VTC-034 GSCs were treated or not with JM2 at different concentrations for 24 h, lysed and analyzed by blotting using Neutravidin-HRP detection, and immunoblotting for GAPDH as loading control. C VTC-034 GSCs were treated with JM2 at 50 μM for different times, lysed and analyzed by blotting using Neutravidin-HRP, and immunoblotting for GAPDH as loading control. D VTC-034 GSCs were treated or not with JM2-scrambled (JM2-scrbl) or JM2 at 50 μM for 24 h, and fixed. Biotin-tagged JM2-scrambled and JM2 were observed by confocal fluorescence microscopy using fluorophore-conjugated streptavidin (red), and DAPI was used to stain nuclei. Scale bar: 10 μm. E Fluorescence confocal microscopy of VTC-037 GSCs treated or not with JM2 at 50 μM for 24 h before fixing, and probed for Wheat Germ Agglutinin (WGA – green), biotin-tagged JM2 (red), with nuclei identified using DAPI (blue). Zoom and 3D projection on right. Scale bar: 10 μm.

Article Snippet: Primary antibodies rabbit anti-Cx43 (MilliporeSigma; C6219), mouse anti-pan Cadherin (MilliporeSigma; C1821), or mouse anti-α-tubulin (MilliporeSigma; T6199) were diluted in blocking buffer and labeling was performed overnight at 4 °C.

Techniques: Binding Assay, Control, Western Blot, Fluorescence, Microscopy, Staining, Confocal Microscopy

Journal: Cell Death & Disease

Article Title: Cytoplasmic connexin43-microtubule interactions promote glioblastoma stem-like cell maintenance and tumorigenicity

doi: 10.1038/s41419-025-07514-2

Figure Lengend Snippet: A Cellular thermal shift assay in VTC-037 GSC lysates was used to determine JM2 and JM2-scrambled selective target engagement potency for α-tubulin and β-tubulin. VTC-037 GSC lysates were subjected to different concentration of JM2 or JM2-scrambled (JM2-scrbl) at 57 °C, and peptide affinity was analyzed by western blotting using antibodies against α-tubulin, β-tubulin, and β-actin as negative control. B Percentage of stabilized α-tubulin and β-tubulin at 57 °C was represented, and EC50 values for JM2 and JM2-scrambled (JM2-scrbl) were calculated. C STORM derived point-cloud localizations of Cx43 (green) and α-tubulin (magenta) in VTC-037 GSCs following treatment with JM2-scrambled (JM2-scrbl) or JM2 at 50 μM for 24 h. Zoomed out panels (left) scale bar: 6 μm. Zoomed in panels (middle) scale bar: 1 μm. Zoomed in panels (right) scale bar: 1 μm. Sphere size: 50 nm. D Cross-Pair correlation functions for Cx43/α-tubulin interaction in C ( n = 10). E VTC-037 GSCs were treated or not with antennapedia (ant), JM2, or JM2-scrambled (JM2-scrbl) at 50 μM for 24 h. Following cross-linking, cell lysates were subjected to co-immunoprecipitation using α-tubulin antibody, or V5 antibody for negative control, and immunoblotted using antibodies against Cx43, α-tubulin, and GAPDH for loading control. Neutravidin-HRP was used to detect biotin-tagged peptides. HC heavy chain, LC light chain.

Article Snippet: Primary antibodies rabbit anti-Cx43 (MilliporeSigma; C6219), mouse anti-pan Cadherin (MilliporeSigma; C1821), or mouse anti-α-tubulin (MilliporeSigma; T6199) were diluted in blocking buffer and labeling was performed overnight at 4 °C.

Techniques: Thermal Shift Assay, Drug discovery, Concentration Assay, Western Blot, Negative Control, Derivative Assay, Immunoprecipitation, Control

Journal: Cell Death & Disease

Article Title: Cytoplasmic connexin43-microtubule interactions promote glioblastoma stem-like cell maintenance and tumorigenicity

doi: 10.1038/s41419-025-07514-2

Figure Lengend Snippet: VTC-001 and VTC-037 GSCs were treated or not with antennapedia (Ant.) or JM2 peptides at 50 μM for 24 h. Cell lysates were analyzed by immunoblotting using antibodies against Notch1 ( A , B ; quantification shown on right), Cx43 ( C , D ; quantification shown on right), and GAPDH as loading control. Statistical analysis was performed with one-way analysis of variance (ANOVA) with Tukey’s multiple comparisons test. ** p ≤ 0.01, *** p ≤ 0.001, ns not significant. Data are represented as mean ± SD. E VTC-037 GSCs were treated or not with JM2-scrambled (JM2-scrbl) or JM2 peptides at 50 μM for 24 h, RNA was extracted and Notch1 mRNA levels were quantified by RT-qPCR. F VTC-037 GSCs were treated or not with JM2-scrambled (JM2-scrbl) or JM2 peptides at 50 μM for 24 h, RNA was extracted and HES1 and HEY1 mRNA levels were quantified by RT-qPCR. G VTC-001 and VTC-037 GSCs were treated or not with antennapedia (Ant.) or JM2 peptides at 50 μM for 24 h, RNA was extracted and HES1 mRNA levels were quantified by RT-qPCR. Statistical analysis in ( E − G ) was performed by two-tailed unpaired Student’s t -test; * p ≤ 0.05, ** p ≤ 0.01, ns not significant. Data are represented as mean ± SD.

Article Snippet: Primary antibodies rabbit anti-Cx43 (MilliporeSigma; C6219), mouse anti-pan Cadherin (MilliporeSigma; C1821), or mouse anti-α-tubulin (MilliporeSigma; T6199) were diluted in blocking buffer and labeling was performed overnight at 4 °C.

Techniques: Western Blot, Control, Quantitative RT-PCR, Two Tailed Test

Journal: Cell Death & Disease

Article Title: Cytoplasmic connexin43-microtubule interactions promote glioblastoma stem-like cell maintenance and tumorigenicity

doi: 10.1038/s41419-025-07514-2

Figure Lengend Snippet: A Following isolation from mouse flank, patient-derived xenograft (PDX) GBM22 cells were cultured as gliospheres or differentiated after addition of 10% FBS ( + FBS) for 24 h, lysed, and analyzed by immunoblotting using antibodies against Cx43, CD133, Olig2, Notch1, SOX2, and GAPDH as loading control. B PDX GBM22 cells were cultured as single cells in suspension in low attachment 96-well plates, and treated or not with 100 μM of JM2-scrambled (JM2-scrbl) or JM2 peptides every other day for 2 weeks. Gliospheres were observed by phase-contrast microscopy. Scale bar: 100 μm. C Quantification of gliosphere size in ( B ) as assessed by diameter measurement ( n = 5). D Quantification of gliosphere numbers in ( B ) ( n = 6). Statistical analysis in ( C , D ) was performed with one-way analysis of variance (ANOVA) with Tukey’s multiple comparisons test. *** p < 0.001, **** p < 0.0001. Data are represented as mean ± SD. E PDX GBM22 were injected in mouse flank, and after 2 days, vehicle/control, JM2-scrambled (JM2-scrbl) or JM2 at 300 μM were administered intratumorally every day for 14 days (created with BioRender.com). F Tumor volume were determined at day 2 before the first administration, and day 15 before the last administration ( n = 7 for each treatment group). A two-tailed unpaired Student’s t -test was used; * p ≤ 0.05, ** p ≤ 0.01, ns not significant. Data are represented as mean ± SD. G After 16 days, tumors were harvested and tumor sections were analyzed for the presence of biotin-tagged JM2-scrambled (JM2-scrbl) and JM2 using streptavidin-conjugated to Alexa Fluor 647 (magenta), and cell death was assessed using TUNEL staining (green). DAPI staining was used to detect nuclei (Scale bar: 500 μm). Zoomed in image from white squares on right (Scale bar: 125 μm). H Hematoxylin and Eosin staining of tumor sections in ( G ). (Scale bar: 500 μm).

Article Snippet: Primary antibodies rabbit anti-Cx43 (MilliporeSigma; C6219), mouse anti-pan Cadherin (MilliporeSigma; C1821), or mouse anti-α-tubulin (MilliporeSigma; T6199) were diluted in blocking buffer and labeling was performed overnight at 4 °C.

Techniques: Isolation, Derivative Assay, Cell Culture, Western Blot, Control, Suspension, Microscopy, Injection, Two Tailed Test, TUNEL Assay, Staining