Fig 1: Downregulation of CX26 and CX43 promotes PEC proliferation and growth. PECs were transfected with si-NC-CX26 + si-NC-CX43, si-CX26 + si-NC-CX43, si-NC-CX26 + si-CX43 or si-CX26 + si-CX43. (A) Cell proliferation was assessed by EdU assay (scale bar, 50 µm). (B) Protein expression of Nanog and Oct4 following transfection, as determined by western blot analysis. (C) Cell apoptosis following transfection, as detected by Annexin V-FITC/PI double staining. (D) Cell cycle analysis following transfection, as detected by PI staining. (E) Colony formation in vitro following transfection. PECs were transfected with NC-CX26 + NC-CX43, CX26 + NC-CX43, NC-CX26 + CX43 or CX26 + CX43 and exposed to hypoxia. Downregulation of CX26 and CX43 promotes PEC proliferation and growth. (F) Cell proliferation following transfection was assessed by EdU assay (×200). (G) Protein expression of Nanog and Oct4 following transfection and hypoxia exposure, as determined by western blot analysis. (H) Cell apoptosis following transfection and hypoxia exposure, as detected by Annexin V-FITC/PI double staining. (I) Cell cycle analysis following transfection and hypoxia exposure, as detected by PI single staining. *P<0.05 vs. si-NC-CX26 + si-NC-CX43 group; #P<0.05 vs. NC-CX26 + NC-CX43 + hypoxia group (n=3). PEC, pulmonary epithelial cell; EdU, 5-ethynyl-2′-deoxyuridine; si, small interfering RNA; NC, negative control; CX, connexin; NC, negative control; PI, propidium iodide.
Fig 2: Downregulation of CX26 and CX43 induces epithelial-mesenchymal transition in PECs. (A) mRNA expression of E-cadherin and N-cadherin in human NSCLC PECs following exposure to hypoxia, as evaluated by RT-qPCR. (B) Protein expression of E-cadherin and N-cadherin in human NSCLC PECs following exposure to hypoxia, as assessed by western blot analysis. (C) Semi-quantification of results from panel B. (D) Migration ability of human NSCLC PECs following exposure to hypoxia, as seen by microscopy (×200). Human NSCLC PECs were transfected with si-NC-CX26 + si-NC-CX43, si-CX26 + si-NC-CX43, si-NC-CX26 + si-CX43 or si-CX26 + si-CX43 and (E) mRNA expression of CX26 and CX43 was evaluated by RT-qPCR, (F) protein expression of CX26 and CX43 was assessed by western blot analysis and (G) semi-quantified, (H) mRNA expression of E-cadherin and N-cadherin was evaluated by RT-qPCR, (I) protein expression of E-cadherin and N-cadherin was assessed by western blot analysis and (J) semi-quantified, (K) migration ability was determined (×200). Downregulation of CX26 and CX43 induces epithelial-mesenchymal transition in PECs. Human NSCLC PECs were transfected with NC-CX26 + NC-CX43, CX26 + NC-CX43, NC-CX26 + CX43 or CX26 + CX43 overexpression vectors, and exposed to hypoxia, then (L) mRNA expression of E-cadherin and N-cadherin was evaluated by RT-qPCR, (M) protein expression of E-cadherin and N-cadherin was assessed by western blot analysis and (N) semi-quantified, and (O) migration ability was determined (×200). *P<0.05 vs. normal group; #P<0.05 vs. si-NC + si-NC group; &P<0.05 vs. the NC-CX26 + NC-CX43 + hypoxia group (n=3). NSCLC, non-small cell lung cancer; PECs, pulmonary epithelial cells; RT-qPCR, reverse transcription-quantitative PCR; si, small interfering RNA; NC, negative control; CX, connexin.
Fig 3: GJB2 expression in healthy controls and psoriasis lesions. (a) Immunohistochemical staining of GJB2 protein in psoriasis lesions. (b) Immunohistochemical staining of GJB2 protein in healthy controls. (c) Quantitative analysis of GJB2 mRNA in healthy controls and psoriasis lesions by qPCR.
Fig 4: Clinical specimen data analysis. Over‐survival curve analyses curves based on the expression levels of GJB2(A), GJB3(B), and both high expression (GJB2 and GJB3) cohorts compared with other cohorts(C) of NSCLC patients using the log‐rank test. (D) mRNA expression level in NSCLC tissue and paired adjacent non‐tumors tissues. Immunohistochemistry of GJB2 (E) in normal lung tissue, LUAD, and LUSC. Immunohistochemistry of GJB3 (F) in normal lung tissue, LUAD, and LUSC (the scale bars of E F pictures are 100 μm).
Fig 5: CX26 and CX43 mediate activation of the P53/MDM2 signaling pathway. (A) mRNA and (B) protein expression of P53, MDM2 and P21 in human NSCLC PECs following exposure to hypoxia, as determined by RT-qPCR and western blot analysis, respectively. (C) Semi-quantification of panel B. (D) mRNA and (E) protein expression of P53, MDM2 and P21 in human NSCLC PECs following transfection with si-NC-CX26 + si-NC-CX43, si-CX26 + si-NC-CX43, si-NC-CX26 + si-CX43 or si-CX26 + si-CX43, as tested by RT-qPCR and western blot analysis, respectively. (F) Semi-quantification of panel E. (G) mRNA expression and (H) protein expression of P53, MDM2 and P21 in human NSCLC PECs following MDC treatment and hypoxia exposure, as evaluated by RT-qPCR and western blot analysis, respectively. (I) Semi-quantification of panel H. *P<0.05 vs. normal group; #P<0.05 vs. si-NC-CX26 + si-NC-CX43 group; &P<0.05 vs. the normal + MDC group (n=3). NSCLC, non-small cell cancer; PECs, pulmonary epithelial cells; RT-qPCR, reverse transcription-quantitative PCR; MDM2, murine double minute-2; si, small interfering RNA; NC, negative control; CX, connexin; MDC, monodansylcadaverine.
Supplier Page from Abcam for Anti-GJB2 antibody