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Figure 4


Fig. 4. Analysis of the role of gelsolin in insulin secretion by RNAi in B1 cells. (A) Western blot showing the effect of RNAi treatment on gelsolin protein levels in B1 cells. GFP-positive B1 cells transfected with either pSUPER-GFP gelsolin RNAi or pSUPER-GFP control plasmids were selected by FACS and used for immunoblotting. Equal loading was confirmed with an anti-actin antibody. (B,C) B1 cells were co-transfected with pSUPER-gelsolin RNAi or pSUPER-control RNAi plasmids and an hGH-expressing vector, and growth hormone secretion was measured after stimulation with 16.7 mM glucose (B), 1 mM IBMX (C, top panels), or a combination of 16.7 mM glucose and 1 mM IBMX (C, bottom panels). Data are mean ± s.e.m. of five independent experiments (B, right panel: *P<4x10-4). (D) Growth hormone secretion in B1 cells co-transfected with pSUPER-gelsolin RNAi or pSUPER-control RNAi plasmids and an hGH-expressing vector after a 2 hour pre-incubation with 10 µM latrunculin B. Data are mean ± s.e.m. of two independent experiments.





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