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. 2024 Dec 10;12:1442192. doi: 10.3389/fcell.2024.1442192

FIGURE 4.

FIGURE 4

Snap29 is required for the polarized transports toward both the rhabdomeres and basolateral membrane. (A) Immunostaining of Snap29RNAiKK108034-, Snap29RNAiGD7222-, Snap29RNAiHMC02467-, or Snap29RNAiJF01883-expressing retina by eyeless–CoinFLP–longGMR–Gal4 using anti-Na+/K+-ATPase-α (green) and anti-Rh1 (blue) antibodies. RFP (red) and asterisks represent the cells expressing RNAi constructs. (B) The ratio of integrated fluorescence density for Rh1 staining of the cytoplasm compared with that of whole cells was plotted. White bars indicate wild-type cells (w), and gray bars indicate the cells expressing RNAi constructs (Ri). Error bars indicate the SD with four retinas. Significance according to two-tailed unpaired Student’s t-test: *p < 0.05. (C) The ratio of integrated fluorescence density for Na+/K+-ATPase-α staining of the cytoplasm in the cells expressing RNAi constructs compared with that in wild-type cells was plotted. White bars indicate wild-type cells (w), and gray bars indicate the cells expressing RNAi constructs (Ri). Error bars indicate the SD with four retinas. Significance according to two-tailed unpaired Student’s t-test: *p < 0.05. (D) Electron micrographs of the ommatidia or photoreceptors expressing Snap29RNAi. Used RNAi lines are indicated. (E) Electron micrographs of the vesicles accumulated in the cytoplasm or Golgi stacks in the cells expressing Snap29RNAi. Used RNAi lines are indicated. Arrowheads and arrows indicate unusual irregular-shaped vesicles and normal vesicles, respectively. (F) Immunostaining of the retina expressing Snap29RNAiJF01883 using anti-Snap29 (green) and Rab6 (blue) antibodies. RFP (red) and asterisks represent the cells expressing Snap29RNAiJF01883. Arrows indicate Snap29 localization on Golgi stacks. Scale bar: 5 μm (A), 2 μm [(D) in ommatidia], 1 μm [(D) in photoreceptors], 500 nm (E), and 5 μm (F).