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. 2022 Mar 15;12(3):455. doi: 10.3390/biom12030455

Figure 3.

Figure 3

Effects of calpastatin, D-cis-diltiazem, Olaparib, and combination treatments on rd1 retinal cell viability. The TUNEL assay labeled dying cells (magenta) in wt and rd1 retinal explant cultures. DAPI (grey) was used as a nuclear counterstain. Untreated wt (a) and rd1 control retina (untr.; b) were compared to retina treated with either 20-µM calpastatin (CAST20, c), 100-µM D-cis-diltiazem (D100, d), 1-µM Olaparib (OLA1, e), or 20-µM calpastatin combined with 1-µM Olaparib (CAST20+OLA1, f). Note the large numbers of dying cells in the rd1 outer nuclear layer (ONL). The scatter plot (g) shows the percentage of TUNEL-positive cells in the ONL. Statistical significance was assessed using one-way ANOVA and Tukey’s multiple comparison post hoc testing performed between the control (rd1 untreated) and 20-μM calpastatin (CAST20), 100-μM D-cis-diltiazem (D100), 1-μM Olaparib (OLA1), and 20-μM calpastatin combined with 1-μM Olaparib (CAST20+OLA1). Except for D-cis-diltiazem, all treatments decreased rd1 retinal degeneration. The combination treatment CAST20+OLA1 did not improve the therapeutic effect seen with the respective single treatments. Untr. wt: 7 explants from 4 different mice; untr. rd1: 27/27; CAST20 rd1: 8/8; D100 rd1: 16/16; OLA1 rd1: 17/17; CAST20+OLA1 rd1: 8/8; error bars represent SD; ns = p > 0.05, ** = p ≤ 0.01, and **** = p ≤ 0.0001. ONL = outer nuclear layer, INL = inner nuclear layer, GCL = ganglion cell layer. Scale bar = 50 µm.