Figure 4: mab21-l3 acts upstream of MCC and ionocyte master regulators. | Nature Communications

Figure 4: mab21-l3 acts upstream of MCC and ionocyte master regulators.

From: mab21-l3 regulates cell fate specification of multiciliate cells and ionocytes

Figure 4

(a–d) mab21-l3 MO decreases foxi1, foxj1 and multicilin expressions. (a–c) Control MO, mab21-l3 MO or mab21-l3 5mis MO (a,c, 30 ng each; b, 40 ng each) was co-injected with fluorescein dextran (100 ng) into the left ventral blastomere at the four-cell stage, and embryos at stage 13 were subjected to whole-mount in situ hybridization. Ventral views of embryos are shown in left six panels, and fluorescence views reveal distribution of injected materials. foxi1 (a), foxj1 (b) and multicilin (c) expressions were decreased in mab21-l3 MO-injected embryos (foxi1, 100%, n=17; foxj1, 81%, n=32; multicilin, 95%, n=21), but not affected in those injected with control MO (foxi1, 100%, n=21; foxj1, 91%, n=35; multicilin, 94%, n=31) or mab21-l3 5mis MO (foxi1, 100%, n=14; foxj1, 100%, n=11; multicilin, 80%, n=30). The middle 12 panels show higher magnification of boxed areas in left 6 panels. The right graphs show the ratio of the foxi1-, foxj1- or multicilin-expressing cell number per field in the MO-injected side to that in the uninjected side. Shown is the average from embryos in two or three independent experiments. n, the number of embryos scored. (d) The indicated MO (60 ng each) was injected into two ventral blastomeres at the four-cell stage, and embryos at stage 13 were subjected to real-time qPCR analysis. Shown is the average of three or seven independent experiments. (e) Mab21-l3 MO enhances cell proliferation. Left panels, the indicated MO (80 ng each) was injected into two left blastomeres at the four-cell stage, and embryos were fixed at stage 13 for phospho-histone H3 staining. Dorsal is upwards. A, anterior; P, posterior. The right graph shows the ratio of the number of phospho-histone H3-positive cells in the injected side to that in the uninjected side. Shown is the average from embryos (control MO, n=15; mab21-l3 MO, n=19) in two independent experiments. (a–e) Error bars represent s.d. *P<0.05, **P<0.01 by Tukey’s test (a–d) or unpaired Welch’s t-test (e). Scale bars, 500 μm (a–c, left 6 panels; e) and 50 μm (a–c, middle 12 panels).

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