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Fura-2 fluorescence is being lost over time due to either photobleaching or leak through cell membrane, the latter being much greater at 37��C than at 25��C (Bourassa 2006). Probenecid, which can reduce the leak, significantly alters fatigue kinetics at 37��C (F. Bourassa and J. M. Renaud, unpubl. data). However, both unstimulated and tetanic ratios remain constant over one hr despite a 80% decrease in the fluorescence measured with [http://www.selleckchem.com/products/tenofovir-alafenamide-gs-7340.html GS-7340] an excitation at 380?nm while fibers were at rest (Bourassa 2006). For this study, all experiments were completed within 1?h for each fiber. Statistical analysis For most of the experiments, ANOVAs were used to determine significant differences. Split plot designs were used because fibers were tested at all-time levels. ANOVA [http://www.selleckchem.com/products/BIBF1120.html BIBF 1120 supplier] calculations were made using the Version 9.3 GLM (General Linear Model) procedures of the Statistical Analysis Software (SAS Institute Inc., Cary, NC). When a main effect or an interaction was significant, the least square difference (L.S.D.) was used to locate the significant differences (Steel and Torrie 1980). The Fishers Exact test was used when proportions of fibers were calculated (Figs. 2A, ?A,4).4). For each variable, a 2?��?4 test was first carried out, the four categories being control, glibenclamide, glibenclamide-verapamil and Kir6.2-/- fibers. If the model gave a significant probability, then individual 2?��?2 tests were carried out to determine significant differences versus control, glibenclamide versus glibenclamide-verapamil, and glibenclamide versus [http://en.wikipedia.org/wiki/Adenylyl_cyclase Adenylyl cyclase] Kir6.2-/-. The word ��significant�� refers only to a statistical difference (P?