Fig. 5.

CAR10 sequestered miR-30 and miR-203 to stabilize SNAI1 and SNAI2. a SNAI1 and SNAI2 expression positively correlated with CAR10 expression in LUAD. Linear regression analysis with the data from datasets GSE19188 and GSE32219 was performed using the Pearson correlation coefficient test. (In linear regression, a Pearson score > 0.3 and P < 0.05 were assumed to indicate significance). b CAR10 and SNAI1/2 expression levels in A549 cells transfected with miR-30 or miR-203 mimics for 48 h were detected by qRT-PCR. Data are presented as the mean ± SEM of three independent experiments, two-tailed Student’s t test. c CAR10 and SNAI1/2 expression levels in A549 cells transfected with miR-30 inhibitor or miR-203 inhibitor for 48 h were detected by qRT-PCR. Data are presented as the mean ± SEM of three independent experiments, two-tailed Student’s t test. d The expression of miR-30 and miR-203 in A549 cells with silenced or overexpressed CAR10 was detected by qRT-PCR. Data are presented as the mean ± SEM of three independent experiments, two-tailed Student’s t test. e Luciferase activity in A549 cells cotransfected with miR-30 and miR-203 and luciferase reporters containing CAR10 or mutant transcript. Data are presented as the relative ratio of firefly luciferase activity to Renilla luciferase activity. Data are shown as mean ± SEM; n = 3 independent experiments, two-tailed Student’s t test. f An RNA pull-down assay followed by biotin-labeled CAR10 and CAR10-antisense to detect miR-30 and/or miR-203 endogenously associated with CAR10. Data are shown as mean ± SEM; n = 3 independent experiments, two-tailed Student’s t test. g AGO2-RIP assay was performed in A549 lysates, followed by qRT-PCR to detect CAR10 and miR-30 or miR-203 associated with AGO2. Data were represented as the mean ± SEM; n = 3 independent experiments. *P < 0.05, **P < 0.01, and ***P < 0.001. NS no statistical significance