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. 2019 Oct 10;52(6):e12699. doi: 10.1111/cpr.12699

Figure 2.

Figure 2

IL‐7 enhanced the sensitivity of NSCLC cells to cisplatin by IL‐7/IL‐7R pathway. A, The mRNA level of IL‐7R in A549/DDP and A549 cells was analysed by qRT‐PCR. B, The levels of IL‐7R in A549/DDP and A549 cells were analysed by Western blot. Densitometry plot of results from the blots is showed. The relative expression levels were normalized to GAPDH. C, Immunofluorescence analysed the protein levels of IL‐7R in A549/DDP and A549 cells. D, E, The A549 and A549/DDP cells were treated with DMSO, cisplatin, IL‐7 or the blocking antibody against the IL‐7 receptor (Ab) alone or combined for 24, 48 or 72 h. The concentration of cisplatin for A549 and A549/DDP cells was 1 and 10 μmol/L, respectively. Cell proliferation analysis using CCK‐8 assay was performed to assess the cell viability after indicated treatment. F, G, The A549 and A549/DDP cells were treated with DMSO, DDP, IL‐7 or the blocking antibody against the IL‐7 receptor alone or combined for 48 h. EdU proliferation assays were performed of A549 and A549/DDP cells after indicated treatment, and the percentage of EdU‐positive cells was quantified. H‐K, Colony‐forming assay was performed to analyse the colony formation efficiency of A549 and A549/DDP cells after indicated treatment. The average numbers of colony were counted. L‐O, The A549 and A549/DDP cells were treated with indicated treatment for 48 h, and the cell apoptosis was measured by flow cytometry. P, Q, Cell apoptosis was evaluated using HOECHST 33342 assay. Representative images of different groups are showed. Scale bars, 20 µm. R, The levels of JAK3, p‐JAK3, STAT5, p‐STAT5, caspase‐3, Bcl‐2 and Bax in A549 and A549/DDP cells after indicated treatment for 48 h were analysed by Western blot. Data represent three independent experiments (average and s.e.m of triplicate samples). The lowercase letters (a, b, c, d) represent statistically significant (P < .05). To compare whether there is a significant difference, same letters marked were considered to have no significant difference between the two groups and different letters marked were considered to have significant difference between the two groups