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. Author manuscript; available in PMC: 2014 Mar 1.
Published in final edited form as: Nat Med. 2013 Aug 25;19(9):1141–1146. doi: 10.1038/nm.3287

Figure 4. TXNIP effects on MafA and insulin production.

Figure 4

TXNIP overexpressing INS-TXNIP cells were analyzed for (a) MafA mRNA by qRT-PCR, (b) MafA protein by immunoblotting and (c) MafA occupancy of the insulin promoter by ChIP and compared to INS-LacZ control cells; IgG control (right panel). (d) Human islets were transfected with CMV-hTXNIP or LacZ and the effects on MafA mRNA expression were assessed by qRT-PCR. Insulin mRNA as assessed by qRT-PCR in (e) INS-TXNIP cells and (f) human islets transfected with TXNIP. (g) Insulin content and (h) insulin secretion were assessed by ELISA in INS-TXNIP and INS-LacZ control cells. (Control LacZ insulin content: 8.1 and secretion: 1.7ng ml−1 per μgDNA) (i) Islet insulin content was measured by ELISA in primary islets of TXNIP-deficient HcB-19 and control C3H mice. (j) Control C3H and TXNIP-deficient HcB-19 mouse islets were transfected with scrambled control or precursor miR-204 and 2 days later insulin content was measured by ELISA and normalized for DNA content. Bars represent means ±SEM of 3 independent experiments; *P<0.05, **P<0.01.