Fig. 4: miR-6236 promotes adipocyte insulin signaling by inhibiting Pten. | Nature Communications

Fig. 4: miR-6236 promotes adipocyte insulin signaling by inhibiting Pten.

From: Myeloid-derived miR-6236 potentiates adipocyte insulin signaling and prevents hyperglycemia during obesity

Fig. 4

a Dual luciferase reporter assay of scrambled control siRNA or miR-6236 binding to wild-type or miR-6236 binding sites-deleted (Mutant) Pten 3′ UTR cloned in pmirGLO plasmid downstream of firefly luciferase (n = 6; WT, p < 0.0001). b PTEN Western blot (n = 2) and quantification (n = 4, normalized data from two independent experiments) in 3T3-L1 cells (Ctl. vs. 6236, p = 0.040; Ctl. vs. siPten, p = 0.0042; Ctl. vs. siPten + 6236, p = 0.0061). c Insulin-stimulated glucose uptake in differentiated 3T3-L1 cells (n = 6) (Ctl. vs. 6236, p = 0.0070; Ctl. vs. siPten, p = 0.0012; Ctl. vs. siPten + 6236, p = 0.0022). d Oil Red O-stained 3T3-L1 cells and quantification (n = 4; scale bar 200 µm; Ctl. vs. 6236, p = 0.0082; Ctl. vs. siPten, p = 0.0019; Ctl. vs. siPten + 6236, p = 0.0050). e PTEN Western blot and quantification in eWAT of DIO wild-type (+/+) and knockout (−/−) male mice (n = 5 mice/group; p = 0.0033). f PTEN Western blot and quantification in wild-type DIO male mice injected i.p. with EVs from knockout or wild-type metabolically-activated BMDMs (n = 5 mice/group). Ctl. scrambled control siRNA, 6236 miR-6236 mimic, WT wild-type, siPten Pten siRNA, i.p. intraperitoneally. Data representative of ≥2 experiments. Data presented as mean ± SEM; ns not significant, *p < 0.05, **p < 0.01, ****p < 0.0001, two-tailed Student’s t test.

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