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. 2016 Dec 6;12(12):e1006474. doi: 10.1371/journal.pgen.1006474

Fig 6. Knock-down of Six1, Sox13, Pim1 and Rreb1 impairs brown adipogenesis.

Fig 6

(A) Oil-Red-O staining of SVF cells isolated from BAT after in vitro differentiation for 7 days. Before adipogenic induction, cells were transfected with two independent (#1 and #2) locked nucleic acid (LNA) longRNA GapmeRs targeting Six1, Sox13, Pim1, or Rreb1. Scrambled (Scr) or Pparg-targeting LNA GapmeRs served as negative and positive controls, respectively. (B) Expression of brown / mitochondrial marker genes was measured by qRT-PCR on day 7 of differentiation and found to be down-regulated upon knock-down of Six1, Sox13, Pim1, Rreb1 and Pparg. (C) Expression of Ucp1 was down-regulated upon knock-down of the indicated genes. (D) Expression of general adipogenic genes on day 7 of differentiation. (E) mRNA levels of the targeted genes were assayed 24-hour post transfection. Panel (B)-(E) summarize the average of three independent experiments. Error bars represent standard deviation. p-values (paired student’s t-test): * <0.05.