Fig. 2: Claudin1 knockdown increases gefitinib sensitivity in NSCLC cells. | Cell Death Discovery

Fig. 2: Claudin1 knockdown increases gefitinib sensitivity in NSCLC cells.

From: Claudin1 decrease induced by 1,25-dihydroxy-vitamin D3 potentiates gefitinib resistance therapy through inhibiting AKT activation-mediated cancer stem-like properties in NSCLC cells

Fig. 2

A PC9/GR and H1975 cells were transfected with siRNA targeting CLDN1 for 24 h and relative mRNA levels were detected by real-time PCR. **P < 0.01. B, C PC9/GR and H1975 cells were transfected with siCtrl or siCLDN1, and viable cells at the indicated time points were counted. **P < 0.01, ***P < 0.001, ****P < 0.0001. D, E siRNA was transfected, after 6 h, various concentrations of gefitinib (0, 3.25, 7.5, 15, 30, 60 μmol/L) were, respectively, co-administered with siRNA for 24 and 48 h. Then the MTT assay was performed to determine cytotoxicity and the IC50 value against gefitinib was calculated (mean ± SD; n = 3; ****P < 0.0001). F, G PC9/GR and H1975 cells were transfected with siRNA, after 6 h, cells were treated with gefitinib (1 μM) for another 48 h and then stained for Edu (Scale bar: 100 μm; original magnification: ×100; representative images from three experiments). Cell proliferation rates were calculated as a percentage of Edu-positive nuclei to total nuclei (mean ± SD; n = 3; ns: not significant, *P < 0.05, **P < 0.01, ***P < 0.001). H, I PC9/GR and H1975 cells were transfected with siRNA, after 6 h, cells were treated with or without gefitinib (1 μM). Treatments were repeated every 3 days. Colony formation was assessed by crystal violet staining. Colony numbers were assessed by using the ImageJ software and clonal formation efficiency was calculated (**P < 0.01, ***P < 0.001, ****P < 0.0001).

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