Fig. 6

Depletion of Flw increases the amount of phosphomyosin in S2R + cells, but leads to a less organized contractile network. A-D) S2R + cells fixed and stained for phosphomyosin in the absence of Fog (left panels), or after the perfusion of Fog (right panels) following treatment with (A) control, (B) Flw, (C) MBS, or MYPT-75D RNAi. Yellow arrowheads denote cells with a coalesced phosphomyosin contractile network in the form of peri-nuclear rings, while cyan arrowheads indicate a more diffuse phosphomyosin network. Yellow boxes denote cells with peri-nuclear rings shown at higher magnification, while cyan boxes denote cells with a diffuse phosphomyosin network shown at higher magnification. Scale bars 10 µm. E–H Quantification of the mean (± SEM) (E) Normalized Fluorescence Intensity, (F & G) Coalesce Index, (H) Number of cells with defined rings for cells following treatment with control (yellow circles), Flw (blue circles), MBS (green circles), and MYPT-75D (peach circles). E Depletion of Flw and MYPT-75D led to a statistically significant increase in normalized mean phosphomyosin fluorescence intensity while the depletion of MBS was no different than that of control RNAi treated samples. F The coalescence index indicated that depletion of Flw and MYPT-75D were statistically indistinguishable from one another but were statistically less organized as compared to MBS and control RNAi treated samples. G In comparing the number of defined rings, Flw depletion led to a lower number of cells with rings as compared to all other RNAi conditions. (**p-value = ,***p-value = , ****p-value < 0.0001, one-way ANOVA with Tukey’s post-hoc analysis, N = 3, N = 29–89 image fields)