Figure 2
Individualization defects in agoTRiPDf testes and Protamine-GFP (green) accumulation in agoTRiPDf spermatids. (a-d) Investment cones visualized by fluorescent microscopy using Texas Red-X phalloidin staining (red) and nuclei with DAPI in control (a,c) and agoTRiPDf testes (b,d). Scattered investment cones with variable size are present in the agoTRiPDf cysts (b,d arrows). (e) The average number of individualization complexes (ICs) in control and agoTRiPDf testis. Error bars indicate mean ± s.e.m. Statistical significance was determined by Welch two-sample t-test. (f-g) Anti-cleaved Caspase 3 (green) highlight the caspase activity in the control and agoTRiPDf cysts. Morphologically normal cystic bulges and waste bags are present and cleaved Caspase3 accumulates in control testes (f). In agoTRiPDf testes (g), the cystic bulges and waste bags are severely damaged. (h–l’) Nuclei are visualized with Protamine-GFP (green) and DAPI (blue), F-actin with Phalloidin-Texas Red (red). Nuclear bundles are slightly disoriented in agoTRiPDf cyst compared to wild type (h,j, i,k). DAPI and Protamine-GFP positive hypercondensed nuclei are present in agoTRiPDf testes (i arrows, l,l’). The chart was created in Microsoft Excel 2016 MSO ver. 16.0.4266.1001 and processed in Adobe Illustrator CS6 ver.
