Increased mitochondrial proline metabolism sustains proliferation and survival of colorectal cancer cells
Fig 7
Selective isoform knockdown reveals the essentiality of both PYCR1 and PYCR2 expression.
A) Representative western blot showing expression of PYCR1 and PYCR2 in HCT116 cells left untreated or transfected for 72 hours with the indicated siRNAs. Actin was used as loading control. The same membrane was blotted with all 3 antibodies PYCR1, PYCR2 and Actin. B) Bar graphs showing number of HCT116 cells 72 hours after transfection with custom siRNAs targeting PYCR1 or PYCR2 and control siRNA (Scr). Untr indicates untreated control cells. C) Representative western blot showing expression of PYCR1 and PYCR2 in RKO cells left untreated or transfected for 72 hours with the indicated siRNAs. Actin was used as loading control. PYCR1 and 2 were assessed in separate nitrocellulose membrane with their respective loading control. D) Bar graphs showing number of RKO cells 72 hours after transfection with custom siRNAs targeting PYCR1 or PYCR2 and control siRNA (Scr). Untr indicates untreated control cells. The bars represent mean ± SD. Data were analyzed using One-way ANOVA with Dunnet multiple comparisons test versus scramble control (n = 3 independent experiments). * p≤0.05, ** p<0.01.