Fig. 3: ROS and JNK are required, but not sufficient to cause BM damage. | Nature Communications

Fig. 3: ROS and JNK are required, but not sufficient to cause BM damage.

From: Basement membrane damage by ROS- and JNK-mediated Mmp2 activation drives macrophage recruitment to overgrown tissue

Fig. 3

a–d In ey > hid,p35 eye imaginal discs, RNAi-induced depletion of ROS-producing Duox (b), transgenic overexpression of a secreted catalase hCatS (c) and inactivation of JNK by expression of dominant negative JNK (JNKDN) (d) results in suppression of BM damage compared with undead controls (a). Single slices focusing on the basal side of DP (top); yellow squares are magnified below. Scale bars, 100 μm (top), 50 μm (bottom). e Quantification of Perlecan intensity in a–d is taken from single slices at the basal surface of DP and PE. Fluorescence intensity at PE was used as internal control for normalization. Data represented as mean fluorescence ± SEM analyzed by one-way ANOVA with Holm–Sidak test for multiple comparisons. ****p < 0.0001, n.s. = no statistical significance. Data from n = 26 (ey > hid,p35), 20 (duox RNAi), 25 (hCatS), and 17 (JNKDN) discs analyzed from four independent experiments. f, g Representative examples of wild-type (ey-Gal4) eye imaginal discs incubated ex vivo in Schneider’s media with 0 μM H2O2 (f) or 50 μM H2O2 (g) labeled for Perlecan (red or gray). Scale bars, 100 μm. h–k Representative examples of wing imaginal discs with wild-type (h, i) or hyper-activated JNKK (hepCA) (j, k) incubated ex vivo in Schneider’s medium with 0 μM H2O2 (h, j) or 50 μM H2O2 (i, k) and labeled for Perlecan. Yellow squares in middle panels are magnified at right. Scale bars, 100 μm. l Quantification of Perlecan intensity in f–k is taken from single slices at the basal surface of DP and PE. Fluorescence intensity at PE was used as internal control for normalization. Data represented as mean fluorescence ± SEM analyzed by one-way ANOVA with Holm–Sidak test for multiple comparisons. n.s. = no statistical significance. Data from n = 14 (ey-Gal4 0 μM H2O2), 16 (ey-Gal4 50 μM H2O2), 4 (salEPv > GFP 0 μM H2O2), 7 (salEPv > GFP 50 μM H2O2), 6 (salEPv > hepCA,GFP 0 μM H2O2), and 15 (salEPv > hepCA,GFP 50 μM H2O2) discs analyzed from three independent experiments. Source data are provided as a Source Data file.

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