Fig. 2

NPM3 promoted WAT browning by stabilizing PRDM16 mRNA. Relative mRNA expressions of PGC-1α, CIDEA and UCP1 after browning induction in 3T3-L1 preadipocytes (3T3-L1 diff) transfected with or without NPM3 overexpression plasmids (NPM3-OE diff) (A) or siRNAs (siNPM3 diff) (B) (n = 3), data were represented as mean ± SEM and analyzed by student t test; *P < 0.05, **P < 0.01, ***P < 0.001; C Glucose consumption in 3T3-L1 preadipocytes after transfected with NPM3 overexpressed plasmids or siNPM3 (n = 3), data were represented as mean ± SEM and analyzed by student t test, *P < 0.05, **P < 0.01; D Relative mRNA expressions of PGC-1α, CIDEA and UCP1 in iWAT with or without CL-316,243/siNPM3 treatment (n = 5), data were represented as mean ± SEM and analyzed by one-way ANOVA followed by Tukey’s test, *P < 0.05, **P < 0.01, ***P < 0.001; E Representative HE and UCP1 immunohistochemical staining in iWAT with or without CL-316,243/siNPM3 treatment, scale bar = 100 μm, the boxed region was shown at higher magnification at right (scale bar = 50 μm); F Western blot to confirm immunoprecipitation (IP) of NPM3; 10% IP cell lysate was used as the input; G Enrichment of selected targets was compared based on NPM3 IP vs. IgG control (n = 3), data were represented as mean ± SEM and analyzed by student t test, *P < 0.05, ***P < 0.001; H qRT-PCR was used to determine the remaining PRDM16 mRNA level compared with the starting time after Actinomycin D treatment, aP2 mRNA was used as a control (n = 3), data were represented as mean ± SEM and analyzed by student t test, *P < 0.05, ***P < 0.001; I Thermogenic marker genes were examined in 3T3-L1 cells transfected with or without NPM3 or siPRDM16 followed by browning induction (n = 3), data were represented as mean ± SEM and analyzed by student t test, *P < 0.05, ***P < 0.001