Extended Data Fig. 2: Nuclear expression of ASS1 is dependent on WT.
From: ASS1 metabolically contributes to the nuclear and cytosolic p53-mediated DNA damage response
a, MC38 cells, colon cancer cells with mutated p53, were incubated with (+) or without Dox (-) for 2 hrs, and after 48 hrs, fractionated into nuclear (Nuc) and cytoplasmic (Cyto) fractions and immunoblotted for ASS1 and p53. Tubulin and H3 were used as a cytoplasmic or nuclear marker, respectively. (n = 3 biologically independent samples). HepG2 WCL was used as a positive control for CPS1 b, LS-174-T cells, colon cancer cells with wild-type p53, were incubated with (Dox) or without Dox (Ctrl) for 2 hrs, and after 48 hrs, fractionated into nuclear (Nuc) and cytoplasmic (Cyto) fractions and immunoblotted for ASS1 and p53. Tubulin and H3 were used as a cytoplasmic or nuclear marker, respectively. (n = 3 biologically independent samples). c, HCT116 WT cells were incubated with (Dox) or without Dox (Ctrl) for 2 hrs, and after 48 hrs, protein was extracted and immunoblotted for ASS1, IPO7 and p53. β-actin was used as a loading control. (n = 3 biologically independent samples) d, HCT116 WT cells were incubated with (+) or without Dox (-) for 2 hrs, and fractionated after 48 hrs. IgG and beads only controls were also immunoblotted for IPO7. (n = 3 independent experiments).
