Fig. 4
Inactivation of Notch signaling reduces cell steatosis in liver cancer cells HepG2 and SK-Hep1. a HepG2 and SK-Hep1 cells were treated with 1 mM serotonin alone and in combination with Notch inhibitors, 2 μM AVG and 1 μM FLI-06 as indicated for 24 h. Fifty micrograms of cell extracts were immunoblotted with anti-cleaved-Notch1V1744, anti-Notch2, anti-Notch3, anti-Jagged1, anti-Hes1, anti-LC3β I/II and anti-GAPDH antibodies. b HepG2 and SK-Hep1 cells were grown on coverslips in 6-well plates and treated with 1 mM serotonin alone and in combination with Notch inhibitors, 2 μM AVG and 1 μM FLI-06 as indicated for 24 h. Cells were further treated with 100 μM OA for another 24 h and stained with Oil Red O stain. Slides were observed under a light microscope using 40X objective and photographed (upper panel). The number of Oil-Red-O stained lipid droplets present in steatotic cells (n = 10) were measured and plotted (lower panel). Data are expressed as the mean ± S.D. *** p < 0.001 compared to control OA; # p < 0.1, # # p < 0.01, # # # p < 0.001 compared to Serot. AVG- avagacestat. Serot. - Serotonin. OA- Oleic acid