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. 2023 Jan 6;150(1):dev201373. doi: 10.1242/dev.201373

Fig. 3.

Fig. 3.

Localized expression of dominant-negative Lv-Ets1 (dnLv-Ets1.GFP) in PMCs disrupts expression of downstream terminal differentiation genes, p16 and sm30b. (A) Schematic of the expression patterns of p16 and sm30b genes. (B) Single-color FISH images showing strong p16 expression at the tips of the growing arms and the tips of the body rods (arrowheads) (see Fig. 2B for treatment schedule). (C) Single-color FISH images showing strong sm30b expression throughout the PMC syncytial network but not in the ventral transverse rods (arrowheads). (D) GFP and p16 immunoFISH staining of transgenic embryos with asymmetric dnLv-Ets1.GFP expression in PMCs. The number of embryos expressing dnLv-Ets1.GFP that exhibited reduced p16 expression in the same region (asterisks) was scored. Expression of dnLv-Ets1.GFP in the ventral transverse rods did not affect p16 expression in the growing arms of the 4-armed pluteus (arrowheads). Top: GFP-immunostained cells. Middle: Cy3-labeled p16 RNA transcripts. Bottom: fluorescence merged with Hoechst 33342 counterstain in grayscale. (E) GFP and sm30b immunoFISH staining of transgenic embryos with asymmetric dnLv-Ets1-GFP expression in PMCs. The number of embryos expressing dnLv-Ets1.GFP that exhibited reduced sm30b expression in the same region was scored. Expression of sm30b is also disrupted in PMCs nearby (asterisks). Top: GFP-immunostained cells. Middle: Cy3-labeled sm30b RNA transcripts. Bottom: fluorescence merged with Hoechst 33342 counterstain in grayscale. Scale bars: 50 μm.