Fig. 4. B4GalT5 knockdown promotes commitment of C3H10T1/2 cells to adipocyte lineage probably through BMP signaling.

a–c The effect of B4GalT5 knockdown on adipogenic commitment and subsequent differentiation was assessed by oil red O staining on day 6 after MDI induction. Mock, blank control; NC, negative control, treated with negative scramble siRNA; T5i, experimental group, treated with B4GalT5 siRNA; BMP4, positive control. a Oil red O staining of SVF cells. b Oil red O staining of adipocyte-progenitor cells. c Oil red O staining of C3H10T1/2 cells. d Western blotting of PPARγ, C/EBPα, and 422/aP2 on day 6 after induction was performed to analyze adipogenesis, protein level of B4GalT5 was also shown. Western blot analysis of the BMP signaling pathways on day 0 using the antibodies indicated above, protein levels of B4GalT5 and Pref1 were also shown, with Hsp90 as a loading control. e On day 6 post-induction, cells were stained with oil red O to assess adipogenesis. f Knockdown effect of smad4 and p38 MAPK were confirmed by western blotting on day 0, and western blotting of PPARγ, C/EBPα, and 422/aP2 on day 6 post-induction. Each data is representative of at least three independent experiments with identical results. Each data is representative of at least three independent experiments with identical results. *P < 0.05, **P < 0.01, ***P < 0.001