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Figure 6.
(A) Schematic overview of the treatments with MEK inhibitor CI1040, PI3K inhibitor LY294002, and anti-inflammatory corticosteroid dexamethasone. Embryos were continuously treated from 48 hr post fertilization (hpf) until 5 days post fertilization (dpf), when either whole-mount in situ hybridization (WISH), confocal imaging, or quantitative reverse transcription PCR (RT-qPCR) was performed. (B,C) WISH staining for the expression of the c-myb and l-plastin markers was scored as low, mid, and high. Measurements originate from at least three distinct experiments. Number on bars: number of embryos. NS, not significant; *p < 0.05, **p < 0.01, ***p < 0.001, chi-squared test. (D) Representative images of 5 dpf Shp2wt and Shp2D61G zebrafish embryos in Tg(mpx:GFP, mpeg:mCherry) background without and with dexamethasone treatment. Scale bars, 150 μm. (E,F) Number of mpx:GFP and mpeg:mCherry-positive cells per embryo. Number on bars: number of embryos. Error bars represent SEM. (G) Expression of tnfa, gcsfb and il1b genes determined by RT-qPCR in Shp2wt and Shp2D61G/D61G zebrafish embryos without and with dexamethasone treatment at 5 dpf, normalized to ef1a expression. Standard deviation of four samples in duplicates for each condition are shown. (E,F,G) *p < 0.05, **p < 0.01, ***p < 0.001. ANOVA complemented by Tukey’s HSD.
Anti-inflammatory treatment of zebrafish Shp2D61G embryos ameliorates the juvenile myelomonocytic leukemia (JMML)-like myeloproliferative neoplasm (MPN) phenotype.