Fig. 6

Effects of MEK6 on adipogenic differentiation and fatty acid metabolism of hAMSCs. a qRT-PCR was used to verify MEK6 expression at indicated time points during adipogenic differentiation of hAMSCs. b MEK6 was silenced using two pairs of siRNAs. c and d qRT-PCR (c) and western blot (d) were performed to analyze adipogenic markers in each group of cells on day 6 after adipogenic induction. e and f Oil Red O staining and quantification were used to verify generation of lipids in each group of cells on day 10 after adipogenic induction. Scale bars: 100 µm. g and h hAMSCs were transduced with lentivirus and overexpression efficiency was detected by fluorescence observation (g) and qRT-PCR (h). Scale bars: 200 µm. i and j qRT-PCR (i) and western blot (j) were used to analyze expression of adipogenic markers in cells on day 6 after adipogenic induction. k and l Generation of lipids in cells on day 10 after adipogenic induction were detected by Oil Red O staining (k) and quantification of Oil Red O staining (l). Scale bars: 100 µm. m Hierarchical clustered heatmap of concentrations of 39 types of FFAs in each group of cells. Each color block represented the concentration value of each FFA in each sample, and the values pre-standardized beforehand. n = 3. n and o Mass spectrometry showed that the relative contents of all significantly down-regulated FFAs in cells with MEK6 knockdown (N) or overexpress (O) and in control cells. p Expression levels of MEK6 in BMI < 25 and BMI ≥ 25 adipose tissue samples by qRT-PCR. q Expression correlation between LPL or PLIN1 and MEK6 in BMI ≥ 25 samples. r Expression correlation between MEK6-AS1 and MEK6 in BMI ≥ 25 samples.GAPDH was used as internal controls for qRT-PCR and western blot. The quantitative data were normalized to GAPDH, n = 3. Data are shown as the mean ± SD. Statistically significant differences were considered as follows: ∗P < 0.05, ∗  ∗P < 0.01, ∗  ∗  ∗P < 0.001, ∗  ∗  ∗  ∗P < 0.0001