Fig. 2: The excessive content of ROS content and hyperactivation of XBP1 drive adipogenesis.

a The schematic diagram illustrating the pivotal role of XBP1s and its downstream factor FASN during adipogenic differentiation and adipose maturation. Two distinct groups of 3T3-L1 preadipocytes were described: those stimulated solely by Inducer I were referred to as Inducer I treated preadipocytes, and those successively stimulated by Inducer I and Inducer II were termed Inducer II treated preadipocytes. b Changes in cellular morphology and (c) intracellular lipid droplet size during (pre)adipocyte adipogenic development, with blue spheres representing intracellular lipid droplets. The time points after Inducer II treatment are labeled as follows: 2 days after treatment as “day 0,” 6 days as “day 4,” and 9 days as “day 7” accordingly. Scale bar = 30 μm. n = 20 lipid droplets examined over three independent experiments. d Representative fluorescence images reflecting intracellular lipid content (scale bar = 100 μm) and (e) ROS level in (pre)adipocytes (scale bar = 30 μm). DCF Dichlorofluorescein. f Representative immunofluorescence images of XBP1s and (g) FASN expression in indicated (pre)adipocytes. Scale bar = 30 μm or 7.5 μm, respectively. h Western blot analysis (i) and quantification (n = 3 in the “Induce II” group and n = 4 in other groups) of XBP1s expression from indicated groups. All experiments were repeated three times independently, with similar results. Source data are provided as a Source Data file. All data was expressed as mean ± SD, ns no significance. Statistical significance was evaluated by an unpaired two-tailed t-test.