Additional file 2 of PHGDH drives 5-FU chemoresistance in colorectal cancer through the Hedgehog signaling

Supplementary Material 2: Supplementary Figure 2. PHGDH level supports CRC aggressiveness traits acquisition. A) PHGDH protein levels in PHGDH-silenced LS174T cells. Vinculin immunoblot was performed to ensure equal loading. Bargraph reports quantification of PHGDH level compared to vinculin and show the mean ± SEM of three independent experiments; t-test ** p<0.01, B) Survival fraction of LS174T cells transfected with shCTRL or shPHGDH after 48h of 5-FU treatment. Data are reported as mean ± SEM from three independent experiments; t-test **p<0.01, **** p<0.0001. C) γH2AX levels in transfected and parental LS174T cells. Vinculin and H2AX were used as loading control. D) Invasive abilities of LS174T cells transfected with shCTRL or shPHGDH. Cells were seeded in the upper compartment of 8 μm Transwell system coated with Matrigel. After 16h invaded cells were stained with Crystal Violet and counted. Representative images of invaded CRC cells are shown below the bargraphs (magnification 20×, scale bar: 100 μm). Data are reported as mean ± SEM from three independent experiments; t-test **** p<0.0001.