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. 2009 Dec 22;107(2):742–747. doi: 10.1073/pnas.0907967107

Fig. 2.

Fig. 2.

ddlc1 mutants have altered caspase activity and autophagy. (A and B) TUNEL assay to detect DNA fragmentation (arrows) in control (A) and ddlc1 mutant (B) salivary glands 2 h AHE (n = 5 animals/genotype). (C) Average caspase activity (±SEM) of pupal lysates collected 4 h after puparium formation and measured by cleavage of fluorogenic caspase-3 substrate Z-DEVD-AMC (n = 3). WT, wild-type Canton S; WT + C.I., Canton S + caspase inhibitor DEVD-CHO; p35, daGAL4 × uas-p35: Ex, precise excision of the P element; ddlc1, ddlc1/Y. (D) Percent animals with salivary gland cell fragments and intact glands in paraffin sections of pupae 12 h AHE. Genotypes: Control (fkhGAL4/+) (n = 16), p35 (fkhGAL4/uas-p35) (n = 13), ddlc1 mutant (ddlc1/Y) (n = 28), and p35 expression in ddlc1 mutant (ddlc1/Y; fkhGAL4/uas-p35) (n = 24). (E and F) Representative fluorescence microscopy images of salivary glands expressing GFP-LC3 (green) and DNA (blue). (E) Control (P-excision; +/+; fkhGAL4/uas-GFP-LC3) salivary glands have large numbers of puncta 2 h AHE (n = 20 glands). (F) ddlc1 mutant (ddlc1/Y; +/+; fkhGAL4/uas-GFP-LC3) salivary glands have few puncta 2 h AHE (n = 17 glands). (G) Average number of GFP-LC3 puncta (±SEM) in the genotypes shown in E and F and in ddlc1 mutant salivary glands 12 h AHE (n = 23 glands). (H) Average number of GFP-LAMP1 puncta (±SEM) in salivary glands of control (P-excision; +/+; +/tub-GFP-LAMP1) 6 h after puparium formation, 2 h AHE, and in ddlc1 mutant (ddlc1/Y; +/+; +/tub-GFP-LAMP1) 2 h AHE, and 12 h AHE (n = 21 glands).