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Fig. 4 Rho GTPases regulate microridge organization.
(A) Larvae at 2.5 dpf were treated with 25 µM Blebbistatin (Bleb) for 1 h to inhibit myosin II. 0.5% DMSO was used as a control. The length of microridges in the same cell before and after treatment was measured. Each data point represents the percentage change of the average microridge length in an individual cell after treatment. Data are representative of at least three different experiments. Each experiment was performed on at least 3 larvae, with measurements performed on 3–4 cells per larva. For details of the methods and analysis, refer to the Methods and Materials section. Efficacy of blebbistatin was confirmed by changes in neutrophil morphology and motility after treatment (data not shown). ns, not significant (two-tailed, unpaired t-test). (Right panel) Representative confocal images of GFP-UtrCH labeled microridges before and after treatment. Scale bar, 10 µm. (B) Maximum intensity projection of epithelial cells expressing Lifeact-Ruby alone (i) or with dominant negative RhoA (Dendra2-RhoA T19N) (ii) in zebrafish larvae at 2.5 dpf. Scale bar, 10 µm. Average distance between microridges (C) and cell area (D) in control or Dendra2-RhoA T19N expressing epithelial cells were measured and compared. Each data point represents the value from one cell. Data are collected from more than ten individual larva for each group from 3 separate experiments. **** P<0.0001 (two-tailed, unpaired t-test). (E) Maximum intensity projection of epithelial cells expressing GFP-UtrCH alone (i) or with dominant negative Rac2 (mCherry-2A-Rac2 D57N) (ii) in zebrafish larvae at 2.5 dpf. Scale bar, 10 µm. Average distance between microridges (F) and cell area (G) in control or mCherry-2A-Rac2 D57N expressing epithelial cells were measured and compared. Each data point represents the value from one cell. Data are collected from more than twelve individual larva for each group from 3 separate experiments. For details of the methods and analysis refer to the Methods and Materials section. **** P<0.0001; ns, not significant (two-tailed, unpaired t-test).