Fig. 5: Mmp2, not Mmp1, is required for BM damage, hemocyte recruitment and AiP. | Nature Communications

Fig. 5: Mmp2, not Mmp1, is required for BM damage, hemocyte recruitment and AiP.

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

Fig. 5

For all panels, scale bars 100 μm. ****p < 0.0001, n.s. = no statistical significance. a–c Single slices focused on the basal side of DP show uniform Perlecan labeling in Mmp2 RNAi discs (c), while Mmp1 RNAi discs (b) still show irregularities as in mock (RFP) RNAi discs (a). d Quantification of Perlecan intensity in (a–c) taken from single slices at the basal surface of DP and normalized to PE, represented as mean fluorescence ± SEM analyzed by one-way ANOVA with Holm–Sidak test for multiple comparisons. n = 26 (mock RNAi), 14 (Mmp1 RNAi), 16 (Mmp2 RNAi) discs analyzed from three independent experiments. e–g Recruitment and activation of hemocytes (NimC in gray) to ey > hid,p35 discs is severely impaired upon knockdown of Mmp2 (g), while Mmp1 RNAi does not affect hemocyte recruitment (f) similar to mock RNAi discs (e). Yellow squares are magnified below. Yellow arrowheads (bottom) indicate activated morphology of hemocytes. h, i Quantification of hemocyte recruitment (h) and activation (i) in ey > hid,p35 discs compared with Mmp1 and Mmp2 knockdown. Hemocyte activation was measured by the percentage of total hemocytes that had one or more cellular protrusions. Total number and percentage of activated hemocytes were analyzed by one-way ANOVA with Tukey’s multiple comparisons test. n = 20 discs per genotype analyzed in three independent experiments. j–l Mmp2 knockdown suppresses adult overgrowth. Representative images of adult heads of ey > hid,p35 animals that are expressing either mock (RFP) RNAi (j), Mmp1 RNAi (k), or Mmp2 RNAi (l). Mmp2 RNAi also suppresses larval imaginal disc overgrowth, as seen in c) and (g, where the disc morphology is restored, in contrast to ey > hid,p35 discs in a and e. m Quantification of the suppression of adult ey > hid,p35 overgrowth by Mmp2 RNAi. Progeny was scored as wild type (WT) (black bars) or overgrown (red bars). Suppression is measured by a shift in percentage to WT from overgrown animals that is significantly different as determined by two-sided Fisher’s exact test. n = 100 to 150 flies counted per genotype from four independent experiments. Source data are provided as a Source Data file.

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