Fig. 7
Ca2+-signalling affects PARP activity, PAR generation, and sirtuin activity. A PARP activity assay (green) was performed in rd1 and wild-type (wt) retinal explant cultures, with DAPI (grey) as nuclear counterstain. Untreated (Untr.) rd1 and wt retina were compared to rd1 retina treated with BAPTA-AM, L-cis, CM4620, SN-6, D-cis, TTA-AS, DS5565, and NA-184. The scatter plot shows percentage of PARP activity positive cells in the outer nuclear layer (ONL). Untr. wt: n = 11; Untr. rd1: 24; BAPTA rd1: 10; L-cis rd1: 5; CM4620 rd1: 5; SN-6 rd1: 7; D-cis rd1: 10; TTA-A2 rd1: 6; DS5565 rd1: 6; NA-184 rd1: 8. B PAR staining (black) was performed in rd1 and wt retinal explant cultures. Untreated wt and rd1 retina were compared to drug-treated retina as in A. Untr. wt: n = 6; Untr. rd1: 17; BAPTA rd1: 7; L-cis rd1: 7; CM4620 rd1: 10; SN-6 rd1: 8; D-cis rd1: 8; TTA-A2 rd1: 8; DS5565 rd1: 8; NA-184 rd1: 9. C Sirtuin activity assay (white) was performed in rd1 and wt retinal explant cultures. Untreated rd1 and wt retina were compared to drug-treated retina as in A. Untr. wt: n = 7; Untr. rd1: 21; BAPTA rd1: 9; L-cis rd1: 9; CM4620 rd1: 10; SN-6 rd1: 8; D-cis rd1: 9; TTA-A2 rd1: 8; DS5565 rd1: 8; NA-184 rd1: 9. Statistical testing: one-way ANOVA with Tukey’s multiple comparison post hoc test performed between rd1 explant cultures. Error bars represent SD; ns = p > 0.05; * = p < 0.05; ** = p < 0.01; *** = p < 0.001; **** = p < 0.0001. INL = inner nuclear layer, GCL = ganglion cell layer; scale bar = 50 µm