Fig. 5

TFRC overexpression partially rescues IGF2BP2 knockdown-mediated attenuation of colorectal cancer cell proliferation and iron metabolism. (A, B) After transfecting TFRC expression plasmids, the mRNA expression of TFRC and downstream factors and cell cycle-related factors were assessed using qRT-PCR. (C, D) Transfection of HCT-116 and SW480 cells expressing shIGF2BP2 with TFRC, and determination of protein levels using western blot. (E, F, G) Cell viability, clonogenicity, and proliferative capacity of shIGF2BP2-expressing HCT-116 and SW480 cells transfected or not transfected with oe-TFRC as assessed using the CCK-8 assay (E), colony formation assay (F), and EdU staining (G), respectively. (H) Assessment of plasmid-transfected HCT-116 and SW480 cells for TFRC via propidium iodide (PI) staining using flow cytometry. (I, J, K) Overexpression of TFRC in HCT-116 and SW480 cells transfected with shIGF2BP2 and determination of reactive oxygen species (ROS) concentration (I), total iron content (J), and labile iron pool (LIP) content (K). Each value represents the mean ± SD for triplicate samples (Student’s t-test). *P < 0.05, **P < 0.01, ***P < 0.001 and **** P < 0.0001