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. 2022 Aug 24;71(11):2360–2371. doi: 10.2337/db22-0205

Figure 4.

Figure 4

miR-130b/301b targets AMPKα expression in adipose progenitors from iWAT. SVF cells were isolated from inguinal white fat from WT mice at the time of weaning. A: Level of P-ACC following beige adipogenic (adipo-) differentiation (6 days). B: The progenitor cells from iWAT of mice were transfected with miR-130b-3p mimics, miR-301b-3p mimics, or negative control (Con). Total (T-)AMPKα protein level was measured at 2 days after transfection. C: Beige adipogenic differentiation was induced at 2 days after transfection of miR-130b-3p mimics, miR-301b-3p mimics, or negative control. At 3 days following beige differentiation, total AMPKα and P-ACC (S79) levels were shown. N = 3–4. *P < 0.05; **P < 0.01. P < 0.05 by one-tail t test. D: Diagram of AMPKα1 3′-UTR sequence targeted by miR-130b-3p and miR-301b-3p. E: The iWAT progenitors were cotransfected with luciferase reporter plasmid containing the AMPK 3′-UTR (position 1728–2226 [includes the miRNA binding sites]), with mimics of miR-130b-3p, miR-301b-3p, or miR-148a-3p. After 48 h, the firefly/Renilla luciferase (Luc) activity was measured. Mean ± SD, n = 9–12 in each group. Same results were confirmed in repeated experiments. *P < 0.05.