Extended Data Fig. 3: ACLY localizes to the nucleus with TGFβ stimulation.
(a) Nuclear fractionation of MEFs stimulated with TGFβ for 24 h. Immunoprobed for total ACLY, pSer455-ACLY (pACLY), fibrillarin as the nuclear loading control and αTubulin as the cytosolic control. n = 3 biological replicates per group. (b) Nuclear fractionation of MEFs stimulated with TGFβ over a time course of 0.5 h to 24 h. Immunoblot for total ACLY, Lamin B1 as the nuclear loading control, and fatty acid synthase (FASN) as the non-nuclear loading control. (c) Representative immunofluorescence images of tissue section from PostniCre, R26-tdT mouse that underwent TAC demonstrate that TdTomato+ activated fibroblasts (red) contain ACLY (green) in nuclei (DAPI, blue). Scale bar = 50 μm. (d) Fusion protein constructs utilized in Fig. 4c. (e) Western blot of MEFs transfected with plasmids containing fusion protein constructs. Panel d created with BioRender.com. Full length western blots in Source Data Fig. 6.
