Figure 5
CNX-011-67 increases both cytosolic and mitochondrial calcium in NIT1 cells and ATP levels in rat islets. Cultured NIT1 cells were loaded with Fluo-3 AM dye for cytosolic calcium measurement (A) and Rhod-2 AM dye for mitochondrial calcium measurement (B) for 60 min. Following a basal measurement, cells were treated with low glucose (2.8 mM), high glucose (16.7 mM) or high glucose (16.7 mM) with 1 μM of CNX-011-67 followed by measurement of Ca2+ levels for 4 min at 6-s intervals. The resulting flux was represented as arbitrary fluorescent units (n = 3). (C) Islet ATP content from sham control and n-STZ animals. (D) Islet IP3 content in normal Wistar rat islets. Islets were preincubated with 2.8 mM glucose for 1 h and later incubated with low glucose (LG, 2.8 mM), high glucose (HG, 11 mM) or high glucose (11 mM) with 1 μM of CNX-011-67 for 60 min for ATP and 5 min for IP3 measurement. ATP (pmol) and IP3 (pg) content are presented per islet. Values represent mean ± S.E.M (n = 4). Statistical analysis was performed by Students t-test (Two-way, unpaired: * P < 0.05 and ** P < 0.01).