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. 2017 Aug 16;22(1):101–110. doi: 10.1111/jcmm.13297

Figure 4.

Figure 4

Effects of Sirt1 on CD38 deficiency‐mediated inhibition of adipocyte differentiation. (A) The protein expressions of Sirt1 and PPARγ. The protein levels of Sirt1 and PPARγ were significantly up‐regulated or down‐regulated, respectively, after stimulation of the adipose differentiation process in CD38−/− MEFs and tubulin was loaded as an internal control. (B–C) Semi‐quantitative analysis of PPARγ and Sirt1 was performed by calculating the density of Western blot bands. (D–F) The mRNA expressions of PPARγ from wild‐type and CD38−/− MEFs treated with various reagents. The mRNA levels of PPARγ were determined by qRT‐PCR from wild‐type and CD38−/− MEFs with EX527 (a Sirt1‐specific inhibitor, 25 μM), Nicotinamide (a Sirt1 non‐specific inhibitor, 10 mM), cADPR (a CD38‐derived metabolite, 0.5 μM) and NAD + (a CD38 substrate, 1.0 mM) for 24 hrs after induction of adipocyte differentiation (D). The relative mRNA expressions of PPARγ in wild‐type (E) or CD38−/− (F) MEFs were compared with control groups when the cells were treated with various stimuli. (G) The mRNA expressions of aP2 in MEFs. The mRNA expressions of aP2 were determined by qRT‐PCR after treatments with nicotinamide, a Sirt1 inhibitor, in WT and CD38−/− MEFs. (H–I) The protein expressions of PPARγ. The protein levels of PPARγ were determined by Western blot analysis with or without resveratrol (50 μM) or nicotinamide (10 mM) in 3T3‐L1 adipocytes, and tubulin was loaded as an internal control. Semi‐quantitative analysis of PPARγ (I) was performed by calculating the density of Western blot bands. (J) The mRNA expressions of PPARγ, aP2 and FASN. The mRNA levels of PPARγ, aP2 and FASN from 3T3‐L1 adipocytes were evaluated by RT‐PCR at day 12 after the initiation of adipogenesis. The values are expressed as the mean ± S.E. from three independent experiments. n = 3. *P < 0.05, **P < 0.01 and ***P < 0.001. vs WT group. # P < 0.05, ## P < 0.01 vs CD38−/− group. [Correction added on 11 September 2017, after first online publication: An incorrect version of Figure 4 was published previously and has been corrected in this version.]