1. Volume 135, Issue 3, Pages 524-534 (October 2008)
Undertaker, a Drosophila Junctophilin, Links Draper-Mediated Phagocytosis and Calcium Homeostasis 
Leigh Cuttell, Andrew Vaughan, Elizabeth Silva, Claire J. Escaron, Mark Lavine, Emeline Van Goethem, Jean-Pierre Eid, Magali Quirin, Nathalie C. Franc 
Cell 
Volume 135, Issue 3, Pages (October 2008)
DOI: /j.cell
Copyright © 2008 Elsevier Inc. Terms and Conditions

2. Figure 1
Phagocytosis of Apoptotic Cell Defect in Df(3R)3-4 Homozygous Embryos
(A and B) Wild-type (A) and Df(3R)3-4 homozygous mutant (B) AO-stained stage 13 embryos (anterior to the left, dorsal side to the top). Insets show higher-magnified views of AO-stained corpses.
(C–F) Macrophages of stage 13 embryos stained with CRQ Ab (green) and 7-AAD (red) that brightly stain apoptotic corpses in wild-type (C), Df(3R)3-4 (D), Df(3R)ED5147 (E), and Df(3R)ED5138 (F) homozygous embryos. The arrow in (D) points to an apoptotic cell seen in close proximity to the mutant macrophage that is not engulfed, as it is not fully surrounded by CRQ staining. Circles indicate single macrophages.
Scale bars represent 10 μm.
(G) A graph of the corresponding PIs ± SD. N, number of macrophages scored. p values are indicated using asterisks.
(H) A schematic of the 82F region of the genome indicating the breakpoints of the deficiencies.
Cell  , DOI: ( /j.cell )
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3. Figure 2
Requirement for CG10233/undertaker in Phagocytosis of Apoptotic Cells
(A and B) Phagocytosis of apoptotic cells by mock- (A) or uta RNAi-treated (B) S2 cells. Scale bars represent 200 μm.
(C) A graph summarizing the quantification of these assays with rab5 RNAi-treated S2 cells as a control. Bars represent the mean percentage of engulfing cells ± standard errors from the mean (SEM) of three independent experiments with duplicated wells.
(D–F) Merged confocal images of yw; +; crq::Gal4, UAS::eGFP (wild-type reference) (D), yw; +; Df(3R)3-4, crq::Gal4, UAS::eGFP (mutant reference) (E), and yw: UAS::CG10233; Df(3R)3-4, crq::Gal4, UAS::eGFP homozygous macrophages (rescue) (F). Apoptotic cells are stained with 7-AAD (red), GFP-expressing macrophages appear green, and CRQ Ab is in blue. Scale bars represent 10 μm.
(G) A graph of the mean PIs ± SD of these macrophages.
Cell  , DOI: ( /j.cell )
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4. Figure 3
UTA Encodes a MORN Repeat-Containing Protein and Colocalizes with PM, ER, and Phagosomal Markers
(A) A schematic of the two isoforms encoded by uta with corresponding MORN repeat amino acid sequences.
(B–D) S2 cells costained with UTA (green) (B) and the Na+/K+ ATPase subunit (red) (C) Abs and the corresponding overlay (D), with DAPI in blue.
(E–G) Embryonic macrophages costained with CRQ (red) (E) and UTA (green) (F) Abs and the corresponding overlay (G).
(H) S2 cells costained with both CRQ (green) and UTA (red) Abs, and DAPI (blue). Of note is that we could not distinguish the staining of UTA on the membrane of the apoptotic cell being engulfed from that of UTA on the phagosomal membrane of the engulfing cells.
(I and J) S2 cells costained with UTA (red) and PDI (green) Abs (I) or with UTA (red) and KDEL (green) Abs (J).
Scale bars in (B)–(D) and (H)–(J) represent 10 μm, and in (E–G) represent 5 μm.
Cell  , DOI: ( /j.cell )
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5. Figure 4
Requirement for the Drosophila Ryanodine Receptor-Encoding Gene, rya-r44F, which Genetically Interacts with uta
(A–C) Macrophages of stage 13 wild-type (A), homozygous rya-r44F16 (B), and rya-r44Fk04913 (C) embryos stained with the CRQ Ab (green) and apoptotic corpses detected with 7-AAD (red). Scale bars represent 10 μm.
(D) A graph of the corresponding PIs ± SD, and of double heterozygous macrophages for the uta deficiency and each rya-r44F allele.
(E) A graph of the changes in fluo-3AM fluorescence in mock, dorai, dstim, and uta RNAi-treated S2 cells exposed to 2.5 μM TG in Ca2+-free medium at 30 s and to 2 mM extracellular Ca s later. Results are given as a fold increase of the mean fluo-3 fluorescence ± SEM measured over time.
Cell  , DOI: ( /j.cell )
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6. Figure 5 Requirement for SOCE in Phagocytosis of Apoptotic Cells
(A–D) Phagocytosis of apoptotic cells (green) by S2 cells (blue) in the absence (A) or presence (B) of 2 mM EGTA, or by dstim (C) and dorai (D) RNAi-treated S2 cells. Scale bars represent 200 μm.
(E) A graph summarizing these assays, where bars represent the mean percentage of engulfing cells ± SEM of three independent experiments in duplicates.
(F–H) Wild-type (F), oraik11505 homozygous mutant (G), and oraik11505/+; Df(3R)3-4/+ double heterozygous macrophages stained with CRQ Ab (green) and 7-AAD (red). Scale bars represent 10 μm.
(I) A graph of the mean PIs ± SD.
Cell  , DOI: ( /j.cell )
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7. Figure 6
drCed-6 and drpr Genetically Interact with uta and rya-r44F and Are Required for Phagocytosis and SOCE
(A–C) Macrophages of stage 13 wild-type (A), homozygous dced6KG03411a (B), and drprrec8Δ5 (C) mutant embryos immunostained with the CRQ Ab (green) and apoptotic corpses detected with 7-AAD (red). Scale bars represent 10 μm.
(D) A graph of the mean PIs ± SD for each double or trans-heterozygous combination compared to homozygous gene mutations.
(E) Changes in fluo-3AM fluorescence of mock, dstim, drCed-6, and drpr RNAi-treated cells. Results are given as a fold increase of the mean fluo-3 fluorescence ± SEM measured over time.
Cell  , DOI: ( /j.cell )
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8. Figure 7 drCed-6 and drpr Are Required for Bacterial Phagocytosis
(A) Graphs summarizing pHrodo E. coli or S. aureus engulfment by RNAi-treated cells for uta, dstim, dorai, draper, and drCed-6. Results are given as a percentage of engulfing cells ± SEM.
(B–G) Abdomens of control w flies (B and E), and drCed-6KG03411a (C and F) and drprrec8Δ5 (D and G) homozygous mutant flies injected with pHrodo E. coli (B–D) or S. aureus (E–G). The bright white spots seen in the abdomen correspond to bacteria that were engulfed by plasmatocytes (adult macrophage-equivalent).
Cell  , DOI: ( /j.cell )
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