Fig. 6

Compromised pronephric structure and ultrafiltration in ELMO1 crispants, but not in PDX1 morphants, can be beneficially reversed upon treatment with pancaspase inhibitor, zVAD-fmk.

Upon treatment with zVAD-fmk (300 μM), pronephric structural defects in ELMO1 crispants at 48 hpf, are significantly restored to normalcy (A,B,D,E); n = 47 for each condition). The pancaspase inhibitor causes no adversity in the control embryos at 48 hpf (D). However, no significant restoration in pronephric structure is observed with zVAD-fmk treatment of PDX1 morphants, at the same age (C,F); n = 47 for both conditions). The white line in image (F) represents the scale bar corresponding to 200 μm. The significant restoration of the glomerular width and neck via zVAD-fmk treatment in ELMO1 crispants as well as the lack of rescue in PDX1 morphants are quantified in graphs (G,H). Altered ultrafiltration in ELMO1 crispants is also improved significantly upon treatment with zVAD-fmk as seen in graph (I) but not in PDX1 morphants, as observed in (J) The n values for each sample set used for the functional assay are as follows: Co. CRISPR (22), Co. CRISPR with zVAD-fmk (23), ELMO1 CRISPR (22), ELMO1 CRISPR with zVAD-fmk (17), Co. Mo. (23), Co. Mo. with zVAD-fmk (23), PDX1 Mo. (26) and PDX1 Mo. with zVAD-fmk (22).