Fig. 2: Knockdown of ILF3 promotes ferroptosis sensitivity in CRC cells.

A Western blot analysis verifying the transfection efficiency of sh-ILF3 in SW620 and DLD-1 cells. B Cell viability of SW620 and DLD-1 cells in the sh-Con and sh-ILF3 groups following RSL3 treatment at different concentrations, assessed using a CCK-8 assay. C MDA levels in sh-Con and sh-ILF3 groups following RSL3 treatment (10 μM, 24 h). D Mitochondrial morphology in sh-Con and sh-ILF3 groups following RSL3 treatment (10 μM, 24 h). E Cell viability analysis of SW620 and DLD-1 cells treated with RSL3 (10 μM, 24 h) alone or in combination with ferrostatin-1 (1 μM, 24 h), Z-VAD-FMK (5 μM, 24 h), 3-methyladenine (2.5 mM, 24 h), or Necrostatin-1 (5 μM, 24 h) using a CCK-8 assay. F DLD-1 cells from the sh-ILF3 and sh-Con groups were cultured on agarose gel plates and treated with RSL3 (10 μM, 24 h) after seven days of culture. Cells were then stained using a Calcein/PI kit and imaged via confocal microscopy. Red represents PI staining, and green represents Calcein staining. G Schematic diagram illustrating the anticancer effect of RSL3 (50 mg/kg) combined with sh-ILF3#3 in a SW620 subcutaneous tumor model. H Tumor volume was measured every three days. I Tumor weight. J Immunohistochemical staining for ILF3 and Ki-67 and 4-HNE/PI co-staining. Scale bar = 20 μm.