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Fig. 4. siRNA-mediated knockdown of Rip11 reduces Rip11 but not FIP2 or RCP mRNA expression and inhibits insulin-stimulated uptake of 2-deoxyglucose. 3T3-L1 adipocytes were electroporated with scrambled siRNA duplex (S) or two different siRNA duplexes directed towards distinct regions of the Rip11 mRNA sequence (A and B, respectively). (A) The level of expression of Rip11, FIP2 and RCP mRNA was measured 48 hours later using quantitative real-time PCR. The level of expression of each mRNA was determined relative to the level of expression of 
2-microglobulin mRNA, and, for each mRNA, this was normalised to the level of expression in the presence of the scrambled mRNA (therefore, equal to 1 in each case). Each bar is representative of determinations arising from five independent preparations of adipocytes, with the data expressed as the mean ± s.e.m. (B) In a parallel experiment, the adipocytes were treated in the absence or presence of 100 nM insulin, as indicated, and 2-deoxyglucose uptake measured in triplicate, as described in Materials and Methods. The panel is representative of two separate experiments, with each bar indicating the mean ± s.d.
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