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. 2023 Nov 6;19(11):e1011010. doi: 10.1371/journal.pgen.1011010

Fig 6. Prox1a puncta in liquid-liquid phase separates regulate PR differentiation.

Fig 6

(A) Schematic highlighting the retinal layers analysed in the zebrafish larvae. (B) Experimental timeline. Larvae were swum in metronidazole (MTZ) for 24 hours at 3dpf (days post-fertilisation), followed by a 2 min treatment with 1,6-Hexanediol (1,6-HD) or 2,5-HD at 6dpf, before processing at 7dpf. (C-D”) Following photoreceptor (PR) ablation, Tg(lws2:nfsb-mCherry, gfap:eGFP) retinas show Prospero homeobox1a (Prox1a, pink) puncta only in the ONL of 2,5-HD control treated retinas (n = 5, 17.86 ±2.3), but not when liquid-liquid phase separates have been disassembled in the 1,6-HD treated samples (n = 7, 10.94 ±1.681, quantified in H. Arrow indicates nuclear Prox1a staining outside of the photoreceptor layer. (E-G”’) zpr1 cone photoreceptors are observed in both control samples (2,5-HD treated ablated retinas and 1,6-HD treated, non-ablated retinas), but much reduced in the ONL of 1,6-HD treated ablated retinas, quantified in I. DAPI labels cell nuclei (blue). Scale bars: 50 μm.