Extended Data Fig. 3: Knockdown efficiency of shFoxk1-1, shFoxk1-2, shFoxk2-1 and shFoxk2-2, and complementation test of FOXK1 and FOXK2.

3T3-L1 adipocytes were transduced to express shRNA targeting Foxk1 and Foxk2. a, Proteins and RNA were prepared and protein levels were determined by western blot using specific antibodies against FOXK1, FOXK2 and β-actin. b, mRNA levels were determined using quantitative real-time PCR, n = 3. Experiment performed once. 3T3-L1 adipocytes were transduced to overexpress FOXK1, FOXK2, shFoxk1 and shFoxk2 in the combinations listed. c, 2-DG uptake was determined under basal and insulin-stimulated conditions (100 nM), n = 4. d, Lactate secreted into medium, n = 4. c, d, Representative experiments out of two are shown. e, Knockdown efficiency and selectivity of shPdk1 and shPdk4 tested in 3T3-L1 adipocytes using quantitative real-time PCR, n = 3. Experiment performed once. f, g, Lactate production in 3T3-L1 adipocytes transduced with shRNA targeting Pdk1 or Pdk4 either alone or in combination with FOXK1 or FOXK2 expression vectors. n = 3. Representative experiments out of two are shown. Data shown as mean ± s.d., n shows biological independent experiments. Unpaired two-sided Student’s t-test; ***P < 0.001, **P < 0.01, *P < 0.05. For gel source data, see Supplementary Fig. 1.