Figure 1
From: Optogenetic regulation of insulin secretion in pancreatic β-cells
Expression of bPAC confers light-induced modulation of [cAMP]i in MIN6 β-cells. (A) Fluorescence micrographs of AdbPAC-transduced cells (MOI = 100) stained for bPAC (antibody against the myc tag) and nuclear DNA (DAPI). Expressed mCherry (epifluorescence) is also shown. Bar: 100 µm. (B) Western blot showing the expression of bPAC (myc tag antibody) and β-actin. [cAMP]i levels are shown for various (C) MOI and (D) durations of pulsed (red bars) or continuous illumination (blue bars). Cells in (C) were illuminated for 10 min continuously. *p < 0.05 vs. dark, #p < 0.05 vs. cells infected at MOI of 10–50. Cells in (D) were infected at MOI = 100. *p < 0.05 vs. illumination for 5–20 min. In (C,D): NT: Non-transduced cells. (E) [cAMP]i levels after 10 min of illumination. Times after termination of the illumination are shown. *p < 0.05 vs. samples at 12–20 min. In (C–E): Dark: AdbPAC-infected cells (MOI = 100) without illumination. Results are shown as mean ± SD from at least three independent experiments, with each measurement carried out in triplicates. Some blots were edited for better (clear cut) representation. The entire blots are included in Supplementary Materials.
