Fig. 2

Ex vivo exposure to NFA or LPA enhances the engraftment efficiency of zebrafish muscle cells in vivo (A) Outline of experimental strategy. ZeMPCs were incubated with lipids for 4 h at 28.5°C, followed by washing out of the media and drugs, harvesting the cells, and splitting into three cell doses (25, 75, or 200 cells/recipient) for transplantation. Treated cells were transplanted into each side of five pre-irradiated casper recipient fish or five non-irradiated prkdc-mutant recipient zebrafish (4–8 months old), followed by imaging of the recipient fish at 7 dpt. (B) Engraftment efficiency as assessed across different compound exposure times. LPA enhances the engraftment efficiency of ZeMPCs treated for 4 h, while NFA enhances the engraftment efficiency after 2 h and 4 h of exposure. (C) The potency of engrafting cells (1/cell frequency) from (B) was estimated with ELDA. Error bars represent the upper and lower confidence intervals (see Table S3 for p values). (D and E) Engraftment efficiency of LPA-treated (D) or NFA-treated (E) ZePMCs assessed across different compound concentrations, as indicated. (F and G) The potency of engrafting cells (1/cell frequency) from (D) and (E) was estimated with ELDA. Error bars represent the upper and lower confidence intervals (see Tables S4 and S5 for p values). DMSO concentration for all groups including experimental controls and as vehicle for NFA and LPA was 0.1% (n = 10 per cell number dose and 30 per treatment; ∗∗p < 0.01, ∗p < 0.05; ns, not significant; limiting dilution assay).