Unraveling the role of ChREBP in lung adenocarcinoma: Expression, regulatory networks, and potential functional impact
Fig 2
Impact of dominant negative ChREBP (dnChREBP) overexpression on colony formation ability and migration in NCI-H1975 LUAD cells.
(A) Schematic representation of doxycycline-inducible vectors used for overexpression of dnChREBP. LTR, long terminal repeat; TRE, tetracycline-responsive promoter element; UBC, human ubiquitin C promoter; rtTA3, reverse tetracycline-transactivator 3; IRES, internal ribosomal entry site; PuroR, puromycin resistance gene; WPRE, Woodchuck hepatitis posttranscriptional regulatory element; SIN LTR, self-inactivating long terminal repeat. (B) Amino acid alignment of human ChREBP-α and ChREBP-β. bHLH, basic Helix-Loop-Helix-Leucine; ZIP, leucine zipper. (C) Outline of the process used to generate dnChREBP-overexpressing NCI-H1975 cells. (D-E) Colony formation assay. Macroscopic visualization of the crystal violet-stained colonies is shown (D). Violin plot showing the intensity distribution of stained colonies formed in dnChREBP-overexpressing NCI-H1975 cells compared to control cells (E). A significant difference (p-value < 0.01) is indicated. (F-G) Transwell migration assay. Representative images of migrated control and dnChREBP cells at 20x magnification is shown (F). The bar graph shows the percentage of migrated cells, with data representing the mean ± SD from three independent experiments (G).