Fig. 2.

CRISPR/Cas9-mediated UCP1 knockout in human adipocytes. UCP1-deficient cells were generated by co-expressing Cas9 and specific sgRNAs targeting UCP1 in SGBS preadipocytes. (A) The cells were differentiated into mature adipocytes. The knockout of UCP1 and the protein levels of adipogenic markers adiponectin (AdipoQ), peroxisome proliferator-activated receptor gamma (PPARG), and perlipin A (PLIN) were confirmed by Western Blot. NC, non-targeting control. (B) Oxygen consumption rates were determined in a plate-based respirometer. ATP: oligomycin-sensitive OCR; leak: oligomycin resistant OCR (C) Basal glucose uptake was determined by ¹⁴C-deoxy-Glucose incorporation. (D) Extracellular acidification rates were determined as in (B). (B-D) Data obtained from both negative controls and UCP1 KO cell types were merged. Data of 3–4 independent experiments are shown as mean + SEM. **, p < 0.01 (Student's t-test).