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. 2022 Nov 19;13:7090. doi: 10.1038/s41467-022-34766-9

Fig. 4. E379K and R212Q impair the adipogenic capacity of PPARγ2.

Fig. 4

a Experimental outline showing the timing of the transduction of PPARγ−/− MEF-CAR cells with adenovirus containing HA-tagged PPARγ2-WT or mutants, and treatment of the transduced cells with the differentiation cocktails (2h-day 3: insulin, dexamethasone, isobutylmethylxanthine, and rosiglitazone, day 3–7: insulin). b Western blot assessing the expression of WT and mutant PPARγ2 at the timepoints 8 h, day 5 and day 7 after adenoviral transduction. The membrane was probed with antibodies against HA-tagged PPARγ and Tubulin (internal control). Three independent experiments were performed, and similar results were obtained. c Oil-red-O staining of lipid droplets at day 7 of differentiation. Three independent experiments were performed and similar results were obtained. Scale bar, 50 µm. d Western blot 7 days after adenoviral transduction. The membrane was probed with antibodies against HA-tagged PPARγ, FABP4, LPL, and Tubulin (internal control). After correction for tubulin and PPARγ levels, relative protein levels for FABP4 were 77% (E379K) and 79% (R212Q) of WT levels, and for LPL 33% (E379K) and 28% (R212Q). Three independent experiments were performed and similar results were obtained. Source data for panel b and d are provided in the Source Data file.