Figure S1. Hemocyte counts in WT and abi... | Rockefeller University Press

Figure S1.

Hemocyte counts in WT and abi5/Df embryos, larval peripheral hemocyte compartments, and LGs, and analysis of Abi subcellular localization. (A) Single confocal sections of peripheral hemocytes from WT and abi5/Df late-third instar larvae. Total peripheral hemocytes were isolated after vortexing larvae with glass beads and stained with DAPI. (B) Number of peripheral hemocytes per larva. n = 12 larvae. (C) Confocal images of primary LG lobes from WT and abi5/Df late-third instar larvae stained with anti-P1 (green) and DAPI (blue). (D and E) Numbers of total DAPI-labeled cells (D) and P1+ plasmatocytes (E) per primary LG lobe. n = 12 lobes. (F) Confocal images of primary LG lobes from WT and abi5/Df late-third instar larvae stained with anti-Antp (green) and DAPI (blue). (G) Numbers of Antp+ posterior signaling center cells in primary LG lobes of WT and abi5/Df late-third instar larvae. n = 10 lobes. (H) Confocal images of whole-mount WT and abi5/Df embryos (stage 17) carrying lz-GAL4 and UAS-mCD8-GFP (dorsal views). Higher magnification views of boxed areas are shown on the right to highlight GFP+ crystal cells clustered around the embryonic proventriculus. (I) Number of GFP+ crystal cells per embryo. n = 10 embryos. (J) Confocal images of peripheral hemocytes from WT and abi5/Df second instar larvae, stained with an anti-Lz antibody (green) and DAPI (blue). Arrowheads indicate Lz+ crystal cells. (K) Percentage of Lz+ crystal cells among all peripheral hemocytes (total DAPI count). n = 10 larvae. (L) Confocal images of whole-mount WT and abi5/Df embryos at stage 17 (lateral view) carrying Srp-GAL4 and UAS-mCD8-GFP, stained with anti-Abi (pseudocolored magenta) and anti-GFP (green) antibodies. (M and N) Confocal images of WT and abi5/Df primary hemocytes carrying HmlΔ-GAL4 and UAS-EGFP (M) or lz-GAL4 and UAS-mCD8-GFP (N), stained with anti-Abi (pseudocolored magenta) and anti-Avl (green) antibodies. Insets show high-magnification views in the cell periphery. Arrowheads indicate Abi+ puncta colocalizing with Avl. Data represent the mean ± SEM. Student’s t test revealed no statistically significant difference (ns). Scale bars: 20 μm (A, C, F, and J); 50 μm (H and L); 5 μm (M and N).

Hemocyte counts in WT and abi ⁵ /Df embryos, larval peripheral hemocyte compartments, and LGs, and analysis of Abi subcellular localization. (A) Single confocal sections of peripheral hemocytes from WT and abi⁵/Df late-third instar larvae. Total peripheral hemocytes were isolated after vortexing larvae with glass beads and stained with DAPI. (B) Number of peripheral hemocytes per larva. n = 12 larvae. (C) Confocal images of primary LG lobes from WT and abi⁵/Df late-third instar larvae stained with anti-P1 (green) and DAPI (blue). (D and E) Numbers of total DAPI-labeled cells (D) and P1⁺ plasmatocytes (E) per primary LG lobe. n = 12 lobes. (F) Confocal images of primary LG lobes from WT and abi⁵/Df late-third instar larvae stained with anti-Antp (green) and DAPI (blue). (G) Numbers of Antp⁺ posterior signaling center cells in primary LG lobes of WT and abi⁵/Df late-third instar larvae. n = 10 lobes. (H) Confocal images of whole-mount WT and abi⁵/Df embryos (stage 17) carrying lz-GAL4 and UAS-mCD8-GFP (dorsal views). Higher magnification views of boxed areas are shown on the right to highlight GFP⁺ crystal cells clustered around the embryonic proventriculus. (I) Number of GFP⁺ crystal cells per embryo. n = 10 embryos. (J) Confocal images of peripheral hemocytes from WT and abi⁵/Df second instar larvae, stained with an anti-Lz antibody (green) and DAPI (blue). Arrowheads indicate Lz⁺ crystal cells. (K) Percentage of Lz⁺ crystal cells among all peripheral hemocytes (total DAPI count). n = 10 larvae. (L) Confocal images of whole-mount WT and abi⁵/Df embryos at stage 17 (lateral view) carrying Srp-GAL4 and UAS-mCD8-GFP, stained with anti-Abi (pseudocolored magenta) and anti-GFP (green) antibodies. (M and N) Confocal images of WT and abi⁵/Df primary hemocytes carrying HmlΔ-GAL4 and UAS-EGFP (M) or lz-GAL4 and UAS-mCD8-GFP (N), stained with anti-Abi (pseudocolored magenta) and anti-Avl (green) antibodies. Insets show high-magnification views in the cell periphery. Arrowheads indicate Abi⁺ puncta colocalizing with Avl. Data represent the mean ± SEM. Student’s t test revealed no statistically significant difference (ns). Scale bars: 20 μm (A, C, F, and J); 50 μm (H and L); 5 μm (M and N).

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