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. 2023 Feb 6;60:102628. doi: 10.1016/j.redox.2023.102628

Fig. 6.

Fig. 6

MARS2 is regulated by ZEB1 in response to Wnt signaling. a. Transcriptional expression of MARS2 was checked by qRT-PCR after TGF-β treatment in A549 cells (n = 9). b. Design of ChIP primers flanking ZEB1 binding sites near the promoter region on MARS2 ORF. c. ChIP assay using ZEB1 ChIP primers on MARS2 gene (n = 6). si-Cont indicates si-control RNA. d. ZEB1 regulation on MARS2 was investigated using anti-ZEB1 siRNA by western blot analysis in A549 cells (n = 3). e. Transcriptional expressions of MARS2 and ZEB1 were analyzed by qRT-PCR upon ZEB1 knockdown using anti-ZEB1 siRNA in A549 cells (n = 9). f. Transcriptional expressions of MARS2 were analyzed by qRT-PCR with canonical Wnt activator 1-Azakenpaullone (Aza) treatment (10 μM for 24 h) in A549 and H1299 cells, respectively (n = 9). g. Transcriptional expressions of MARS2 were analyzed by qRT-PCR with canonical Wnt inhibitor IWP-2 treatment (30 μM for 24 h) in A549 and H1299 cells, respectively (n = 9). h. Wnt regulation on MARS2 expression was investigated with Aza and anti-ZEB1 siRNA treatments in A549 cells by immune-fluorescence microscopy (n = 3). i. Comparison of MARS2 expression levels in 4 normal lung cells (IMR90, MRC-5, primary small airway epithelial cells, and WI-38) and 12 lung cancer cell lines (A549, Calu-1, Calu-3, ChaGo-k1, EKVX, HOP92, H322, H322M, H460, H520, H522, H1299) (n = 3). All the quantitative data in graphs are marked as the mean ± S. D from at least three independent samples. Statistical analyses of results were performed with Student's t-test. (**, P < 0.01, ***, P < 0.001).