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. 2018 Jul 9;67(10):1999–2011. doi: 10.2337/db17-1174

Figure 7.

Figure 7

Rp-8-Br-cAMPS-pAB alters the G-actin (G)/F-actin (F) ratio. INS-1 832/13 cells were incubated overnight in low-serum and low-glucose culture media and were further incubated for 1 h with low-glucose KRBH prior to each experiment. Cells were then pretreated for 20 min with low-glucose KRBH containing the vehicle, 4-Abn-OH (10 μmol/L), or Rp-8-Br-cAMPS-pAB (10 μmol/L). Next, cells were exposed for 5 min to KRBH containing low glucose (1 mmol/L) or high glucose (20 mmol/L) prior to lysis in a buffer that solubilizes G-actin but not F-actin. Centrifugation was used to pellet F-actin while leaving G-actin in the supernatant. Proteins were subjected to 10% SDS-PAGE and Western blot analysis to detect G- and F-actin. Densitometry analysis of three independent experiments is shown as the mean ± SEM. The y-axis scaling indicates the percentage of total actin; *P < 0.05 vs. 1 mmol/L glucose in the presence of vehicle; **P < 0.05 vs. 20 mmol/L glucose in the presence of vehicle.