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. 2005 Oct 10;391(Pt 2):291–300. doi: 10.1042/BJ20050468

Figure 4. Glucose stimulates an increase in ribosome recruitment on to ribosome-associated proinsulin mRNA.

Figure 4

(a) MIN6 cells were preincubated in KRB containing 2 mM glucose for 1 h followed by incubation in KRB containing 2 or 20 mM glucose for a further period of 1 h. Cells were lysed and polysome analysis was carried out using 7–47% sucrose gradients. The gradients were fractionated from left (fraction 1) to right (fraction 20). (ai) Absorbance of the gradients was measured continually at 254 nm to give polysome profiles. (aii) RNA was isolated from each fraction and run on a 1% agarose formaldehyde gel. RNA was transferred on to a nylon membrane and probed for proinsulin (PI), CPH, PC2, actin and S7 mRNAs. The results presented are representative of three separate experiments. (b) Cells were treated and fractionated as described in (a) except that 15 mM EDTA was included in the lysis buffer and the sucrose gradients to dissociate the ribosomes.