Fig. 6: MOF regulates fat storage by regulating the Glut4 transcription network.

a, b, d, f Left: representative images of neutral lipid staining (BODIPY-493; green). Scale bars: 5 µm. Right: Boxplots showing quantification of overall lipid droplet area per droplet (µm²) in logarithmic scale with the boxes depicting interquartile range. Number of biological replicates per condition: n = 3. Total number of droplets used for quantification are indicated in the figure. Dashed horizontal lines mark the mean area of controls. a Control, Mof-iAdipo or Mg149-treated (50 nM) control cells at baseline (“-”, upper panel) or after 15 min of insulin/glucose challenge (“+”, lower panel). Statistical analysis was performed using the raw data and one-way ANOVA followed by Kruskal–Wallis comparison test, ****p = 10⁻¹⁶. b Control and Mof-iAdipo cells treated with Ex-527 (200 nM) after insulin/glucose challenge. Statistical analysis was performed using the raw data and one-way ANOVA followed by Kruskal–Wallis comparison test, ***p = 0.006. See Supplementary Fig. 7a for baseline images. c Scatter-plot depicting the H4K16ac MFI in control and Mof-iKO iAdipo with (open circles) or without (filled circles) insulin and glucose challenge. Each dot represents a single iAdipo. Statistical analysis was performed using the raw data and two-sided one-way ANOVA followed by Kruskal–Wallis comparison test, **p = 0.014, ****p = 10⁻¹⁶. Number of biological replicates n = 4. d Control and Mof-iAdipo cells at baseline, following overexpression of Glut4, or following ectopic expression of wild-type Mof (wt-Mof) or Mof catalytic mutant (Mof-E350Q) challenged with insulin/glucose (+Ins/Gluc). Statistical analysis was performed using the raw data and two-sided one-way ANOVA followed by Dunn’s multiple comparison test, ***p = 0.001. Note that the data for untreated wild type (n = 1299) and Mof-iKO (n = 4644) is identical to the data depicted in (a). e Heatmap showing the H4K16ac levels from wild type and Mof-iKO upon ectopic expression of Glut4, wt-Mof or Mof-E350Q with or without glucose and insulin treatment. Statistical analysis was performed using the raw data and two-sided two-way ANOVA followed by Sidak multiple comparison post test, ****p = 10⁻¹⁶. f Control and Mof-iAdipo cells treated with chloroquine at baseline or upon insulin/glucose challenge. Statistical analysis was performed using the raw data and two-sided one-way ANOVA followed by Dunn’s multiple comparison test, ***p = 0.0001, ****p = 10⁻¹⁶. g RT-qPCR analyses of visceral WAT. Violin plots show the mRNA expression of Glut4, Pparg, Pcg1α and Mef2c as an average of biological replicates (Mof^+/+ n = 5; Mof^+/− n = 5 for Glut4, Pparg, Pcg1α and Mof^+/+ n = 3; Mof^+/− n = 3 for Mef2c). Statistical analysis was performed by two-sided Mann–Whitney test, *p = 0.026. Dotted lines show the quartiles and solid lines depict the medians. h ChIP-qPCR analyses of MOF at promoters of Pparg, Glut4, Pcg1α and Mef2c promoters in visceral WAT. The data are expressed as fold change over H3. Violin plots show the average of biological replicates (Mof^+/+ n = 6 and Mof^+/− n = 5). Statistical analysis was performed by two-sided Mann–Whitney test, *p = 0.015. Dotted lines show the quartiles and dashed lines depict the medians. i ChIP-qPCR analyses of MOF (left) and H4K16ac (right) at promoters of Pparg in the presence or absence of Glu/Ins in wild-type and Mof-iKO adipocytes. Dotted lines inside the violin plots show the quartiles and dashed lines depict the medians.