Supplementary Figure 8: Both nuclear PDC and mitochondrial PDC are required for prostate cancer cell proliferation. | Nature Genetics

Supplementary Figure 8: Both nuclear PDC and mitochondrial PDC are required for prostate cancer cell proliferation.

From: Compartmentalized activities of the pyruvate dehydrogenase complex sustain lipogenesis in prostate cancer

Supplementary Figure 8

a-g) Representative images of confocal microscopy for PDHA1 and the N-terminus mature form of SREBF1 (n = 3, independent cell cultures, 5 fields acquired for each group and scale bar represents 10 μm) (a), western blot analysis of indicated proteins (n = 3) (b), representative images of PDHA1 and lipid droplets in confocal microscopy. Uncropped images are in Supplementary Figure 15. (n = 3, independent cell cultures, 5 fields acquired for each group and scale bar represents 50 μm) (c), evaluation of tumour formation in xenotransplantation experiments (n = 6, independent tumour samples) (d), tumour mass in xenotransplantation experiments (n = 6, independent tumour samples) (e), representative micrographs in histopathological analysis (haematoxylin/eosin staining and indicated proteins) in xenotransplantation experiments (n = 6, independent tumour samples, 5 fields acquired for each group and scale bar represents 50 µm) (f) and quantification of the percentage of Ki-67 positive cells in xenotransplantation experiments (n = 6, independent tumour samples) (g) of shPDHA1 or shRNA control 22Rv1 cells infected with NES-PDHA1, NLS-PDHA1 alone or in combination. (h) Western blot analysis of indicated proteins in the 22Rv1 cells infected with two shPDHA1 and a scramble shRNA control and treated with SB-204990, a selective ACLY inhibitors at the dose of 30 μM which effectively arrest cell growth as shown in panel (i) in this figure or vehicle for 9 days (n = 3, independent cell cultures). Uncropped images are in Supplementary Figure 15. (i,j) the relative cell number quantification of the 22Rv1 cells (i) and PC3 cells (j) infected with two shPDHA1 and a scramble shRNA control and treated with SB-204990, a selective ACLY inhibitors at the dose of 30 μM for 9 days (n = 3, independent cell cultures). Error bars indicate s.e.m. *P < 0.05; **P < 0.01. n.s, not significant.

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