Figure 5.
Macrophage Nrf2 delays acquisition of early senescence-like features and limits collateral damage. (A) stage 15 Nrf2RNAi macrophages activated JNK signaling (TRE-GFP reporter, green) above control levels. (B) Apoptotic Cleaved Caspase 3 (CC3) in stage 15 control and srp > Nrf2RNAi macrophages. (C) The number of ventrally localized srp > Nrf2RNAi macrophages was indistinguishable from wild type. (D and E) Stage 15 srp > Nrf2RNAi macrophages upregulated the p21/p27 homolog Dacapo (magenta). (F and G) Stage 15 Nrf2RNAi macrophages are characterized by larger nuclei (F) and reduced levels of B-type Lamin Dm0 (G). (H) Levels of hydrogen peroxide (H2DCFDA) were increased in the epithelium upon macrophage Nrf2RNAi. (I and J) Levels of hydrogen peroxide (H2DCFDA) were reduced in the epithelium of H99 embryos but activation of GstD1, ARE-GFP reporter was unchanged. (K and L) DNA oxidation (8oxodG) was significantly increased in the epithelium of srp > Nrf2RNAi embryos. (M) The H99 mutation rescued the increase in the levels of epithelial DNA oxidation (8oxodG) caused by macrophage Nrf2RNAi to subcontrol levels. (N) DNA oxidation (8oxodG) was increased in the epithelium of srp > Nrf2RNAi embryos when compared with embryos carrying the RNAi construct alone (UAS-Nrf2RNAi). (O and P) Levels of the senescence marker p21/p27 Dacapo were significantly increased in the epithelium of srp > Nrf2RNAi embryos. Cell bodies indicated by white dashed outline. Macrophages labeled in green (srp > GFP, D, E) or red (fascin, A); adjacent epithelium is marked by yellow dashed lines (D and E). Data (A–D, F–H, K, and M–O) generated using Nrf2RNAi Flybase ID FBtp0069370 and data (E, L, and P) generated using Nrf2RNAi TRiP HMS02021. ns: not significant, *, P < 0.05; **P < 0.01, ****P < 0.0001 via Mann-Whitney test (A′, B, D′, E, F′, G, H, J, K, O′, P) or unpaired t test (C, I, and N), one-way ANOVA followed by Dunn’s comparison analysis (M). Images collected from 5 controls and 6 srp > Nrf2RNAi embryos (A); 15 control, 15 srp > Nrf2RNAi (B); 5 controls, 6 srp > Nrf2RNAi (D); 5 controls, 7 srp > Nrf2RNAi (E); 6 controls, 8 srp > Nrf2RNAi embryos (F–G).

Macrophage Nrf2 delays acquisition of early senescence-like features and limits collateral damage. (A) stage 15 Nrf2RNAi macrophages activated JNK signaling (TRE-GFP reporter, green) above control levels. (B) Apoptotic Cleaved Caspase 3 (CC3) in stage 15 control and srp > Nrf2RNAi macrophages. (C) The number of ventrally localized srp > Nrf2RNAi macrophages was indistinguishable from wild type. (D and E) Stage 15 srp > Nrf2RNAi macrophages upregulated the p21/p27 homolog Dacapo (magenta). (F and G) Stage 15 Nrf2RNAi macrophages are characterized by larger nuclei (F) and reduced levels of B-type Lamin Dm0 (G). (H) Levels of hydrogen peroxide (H2DCFDA) were increased in the epithelium upon macrophage Nrf2RNAi. (I and J) Levels of hydrogen peroxide (H2DCFDA) were reduced in the epithelium of H99 embryos but activation of GstD1, ARE-GFP reporter was unchanged. (K and L) DNA oxidation (8oxodG) was significantly increased in the epithelium of srp > Nrf2RNAi embryos. (M) The H99 mutation rescued the increase in the levels of epithelial DNA oxidation (8oxodG) caused by macrophage Nrf2RNAi to subcontrol levels. (N) DNA oxidation (8oxodG) was increased in the epithelium of srp > Nrf2RNAi embryos when compared with embryos carrying the RNAi construct alone (UAS-Nrf2RNAi). (O and P) Levels of the senescence marker p21/p27 Dacapo were significantly increased in the epithelium of srp > Nrf2RNAi embryos. Cell bodies indicated by white dashed outline. Macrophages labeled in green (srp > GFP, D, E) or red (fascin, A); adjacent epithelium is marked by yellow dashed lines (D and E). Data (A–D, F–H, K, and M–O) generated using Nrf2RNAi Flybase ID FBtp0069370 and data (E, L, and P) generated using Nrf2RNAi TRiP HMS02021. ns: not significant, *, P < 0.05; **P < 0.01, ****P < 0.0001 via Mann-Whitney test (A′, B, D′, E, F′, G, H, J, K, O′, P) or unpaired t test (C, I, and N), one-way ANOVA followed by Dunn’s comparison analysis (M). Images collected from 5 controls and 6 srp > Nrf2RNAi embryos (A); 15 control, 15 srp > Nrf2RNAi (B); 5 controls, 6 srp > Nrf2RNAi (D); 5 controls, 7 srp > Nrf2RNAi (E); 6 controls, 8 srp > Nrf2RNAi embryos (F–G).

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