Fig. 4: SREBP1c is essential for PHF2 loss-induced lipogenesis and cell proliferation.

a–g Hep3B cells were transfected with the indicated siRNAs. a Proteins were analyzed using western blotting. n = 3 independent experiments. Source data are provided as a Source Data file. b Total FFA levels of cells were measured; mean ± SD (n = 3 independent samples). *P < 0.05. c The expression of lipogenesis- and proliferation-related genes were analyzed by RT-qPCR. mRNA levels were quantified relative to 18S RNA levels; mean ± SD (n = 3 independent experiments). *P < 0.05. d Representative images of Nile Red stained cells. Hep3B cells stained by Nile Red and DAPI were visualized by fluorescence microscopy. n = 3 independent experiments. Scale bar = 60 µm. siP2: siPHF2. e FACS analysis of Nile Red stained cells. Numbers indicate the mean fluorescence intensity. f The colony formation assay of Hep3B cells. Hep3B cells (1 × 10⁴ cells/well) were seeded in six-well plates. Images were acquired after fixation with methanol and staining with 0.5% crystal violet. Scale bar = 4 mm. Colony numbers were counted; mean ± SD (n = 3 independent samples); *P < 0.05. g Representative light micrographs of spheroid formation of Hep3B cells cultured on oxygen-permeable chips for 1 and 5 days after seeding. Scale bar = 200 µm. The average spheroid diameter on Oxy chips was quantified using ImageJ. Mean ± SD (n = 3 independent experiments); *P < 0.05. For the analyses in (b, c, f, g), an unpaired two-tailed Student′s t test was conducted. The exact p values are shown in Supplementary Data 2.