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. 2019 Dec 27;9:20152. doi: 10.1038/s41598-019-56817-w

Figure 6.

Figure 6

Lactate induces browning of 3T3-L1 adipocytes by enhancing PPARγ activity. (A) IP analysis using an anti-PPARγ antibody in day 9 3T3-L1 adipocytes cotransfected with PPARγ, PGC-1α, and RIP140 expression plasmids following 24 h treatment with control bacterial culture medium (con), LKU4-CM, or 5 mM lactate (Cropped blots were used). (B) ChIP assays using the indicated antibodies or normal IgG in day 7 3T3-L1 adipocytes cotransfected with PGC-1α and RIP140 expression plasmids in the presence or absence of LKU4-CM or 5 mM lactate, as indicated. The Ucp1 promoter containing a PPRE was PCR-amplified from immunoprecipitates. con vs LKU4-CM, Lactate; **P < 0.005 (C) Relative Ucp1 promoter activity in HEK293T cells cotransfected with pGL3-Ucp1-Luc and indicated expression plasmids or siRNA in the presence or absence of lactate (5 mM). (D,E) RT-qPCR analysis of Ucp1 (D) and mitochondrial DNA (mtDNA) copy number and citrate synthase (CS) activity (E) in day 8 3T3-L1 adipocytes cotransfected with the indicated expression plasmids or siRNA after 48 h of treatment with vehicle or lactate. (F) Day 8 3T3-L1 adipocytes were incubated with vehicle (ctrl) or lactate for 24 h and the OCR was then measured in the basal condition and in the presence of oligomycin, FCCP, and rotenone/antimycin A at the indicated time point (C–E *P < 0.05, **P < 0.005).