ZFIN Figure: Karampelias et al., 2022, Fig. 3

Fig. 3

CID661578 targets Mnk2b in vivo to promote β-cell regeneration.

a–e, Representative images of Tg(ins:H2BGFP); Tg(ins:flag-NTR) larvae treated with MTZ from 3 to 4 d.p.f. to ablate β-cells, followed by treatment with DMSO (a), 10 µM CID661578 (b), 500 nM cercosporamide (c) or a combination of drugs thereof (d) for 2 d; scale bars, 10 µm. Quantification of the regenerated β-cells is shown in e; n = 15 (control), n = 14 (CID661578), n = 14 (cercosporamide) and n = 14 (CID661578 + cercosporamide). A Kruskal–Wallis test followed by Dunn’s multiple comparisons test was used to assess significance for e (**P = 0.0027, *P = 0.0280 and *** P = 0.0003). Data are presented as mean values ± s.e.m. f–i, Representative maximum projections of mknk2b^+/+ (f), mknk2^+/– (g) and mknk2b^−/− (h) Tg(ins:H2BGFP); Tg(ins:flag-NTR) 6 d.p.f. larvae. Quantification of the β-cell number in the basal state for all genotypes is shown in i; scale bars, 10 µm; n = 7 (mknk2^+/+), n = 23 (mknk2b^+/–) and n = 6 (mknk2b^–/–). Data are presented as mean values ± s.e.m. j–m, Representative maximum projections of mknk2b^+/+ (j), mknk2b^+/– (k) and mknk2b^–/– (l) Tg(ins:H2BGFP); Tg(ins:flag-NTR) 6 d.p.f. larvae following 2 d of β-cell regeneration. Quantification of the β-cell number for all genotypes is shown in m; scale bars, 10 µm; n = 11 (mknk2b^+/+), n = 18 (mknk2b^+/–) and n = 6 (mknk2b^–/–). A Kruskal–Wallis test followed by Dunn’s multiple comparisons test was used to assess significance for m (*P = 0.0198). Data are presented as mean values ± s.e.m. n–r, Single-plane confocal images of Tg(ins:H2BGFP); Tg(ins:flag-NTR) larvae treated with DMSO (n–q) or CID661578.6 (n′–q′) that were uninjected (n and n′) or injected at the one-cell stage with control fabp10a:H2BmCherry (o and o′), tp1:mknk2b (p and p′) or tp1:Hsa.MKNK2 (q and q′) vectors together with transposase mRNA to induce mosaic overexpression of the zebrafish Mnk2b or the human MNK2 in Notch-responsive cells. Quantification results revealed that overexpression of either mknk2b or MKNK2 significantly blocked the effect of CID661578.6 on β-cell regeneration (r); scale bars, 10 µm; n = 15 (control + DMSO), n = 13 (control + CID661578.6), n = 9 (fabp10a:H2BmCherry + DMSO), n = 7 (fabp10a:H2BmCherry + CID661578.6), n = 13 (tp1:mknk2b + DMSO), n = 17 (tp1:mknk2b + CID661578.6), n = 14 (tp1:Hsa.MKNK2 + DMSO) and n = 17 (tp1:Hsa.MKNK2 + CID661578.6). A one-way ANOVA followed by Tukey’s multiple comparisons test was used to assess significance for r (****P < 0.0001 for control + DMSO versus control + CID661578.6, control + CID661578.6 versus tp1:mknk2b + CID661578.6 and control + CID661578.6 versus tp1:Hsa.MKNK2 + CID661578.6). Data are presented as mean values ± s.e.m.

Source data

Expression Data

Expression Detail

Antibody Labeling

Phenotype Data

Phenotype Detail

Acknowledgments

This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Nat. Chem. Biol.