Fig. 4.

GATA6 and TSPAN12 play a crucial role in NSCLC progression. (A) qRT-PCR measures GATA6 and TSPAN12 mRNA levels in lung cancer cells transfected with shGATA6 or shTSPAN12 plasmids. (B) Western blot analysis measures GATA6 and TSPAN12 protein levels in lung cancer cells transfected with shGATA6 or shTSPAN12 plasmids. (C) Cell proliferation assay for GATA6 or TSPAN12 knockdown H1299 lung cancer cells. The cell growth rates were measured by cell counting kit 8. The values present mean ± SD as determined by quintuplet assays. (D) Cell migration assay for GATA6 or TSPAN12 knockdown H1299 cells using transwell membranes. The average counts were derived from six random microscopic fields. (E) Proportion of cells in each cell cycle phase in H1299/shGATA6, H1299/shTSPAN12, and H1299/shCont cells were determined by flow cytometry analysis. (F) Western blot analysis of cell cycle G1 phase-related proteins (Cyclin D1, Cyclin D3, CDK4, CDK6, P18, and P21) in H1299/shTSPAN12 cells and control cells. The bands were quantified using ImageJ software and relative values were obtained by normalizing to the value of each corresponding GAPDH. (G) Expression level of GATA6 in 60 paired NSCLC tissues and their matched NATs. The RNA samples were extracted from 30 NSCLC tissues and 30 corresponding NATs. The RNAs were subject to qRT-PCR with a GATA6 probe and the expression was normalized by GAPDH. (H) Expression level of TSPAN12 in 60 paired NSCLC tissues and their matched NATs. The RNA samples were extracted from 30 NSCLC tissues and 30 corresponding NATs. The RNAs were subject to qRT-PCR with a TSPAN12 probe and the expression was normalized by GAPDH.