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. 2017 Mar 8;7:118. doi: 10.1038/s41598-017-00118-7

Figure 3.

Figure 3

ARF1 and Asrij negatively regulate Toll pathway- mediated immune response. (A,B) Quantification of Toll pathway-governed antimicrobial peptide expression by qRT-PCR analysis shows that Drosomycin and Metchnikowin are upregulated and Defensin is downregulated in arf1 knockdown (A) and asrij null mutant (B) larvae. (C–F) Quantification for the total percentile of flies expressing the Toll pathway reporters - Drosomycin-GFP and Defensin-GFP in flies with e33cGal4-mediated arf1 knockdown (C,D) or asrij null (E) or e33cGal4-mediated asrij knockdown (F) respectively. (G) Images showing increased colocalization of Cactus and Ubiquitin in arf1 or asrij knockdown circulating larval hemocytes as compared to respective controls, also indicated by adjacent co-localization plots. (H) Images showing higher Dorsal specific signal in the entire hemocyte as well as in DAPI stained region (nucleus) for the asrij null (arj9/arj9) and arf1 knockdown (HmlGal4, UASGFP; UAS arf1rnai) larvae as compared to the respective controls (w1118 and HmlGal4, UASGFP). White dotted line indicates nuclear area under consideration. Arrowheads mark nuclei with higher Dorsal signal (I,J) Quantification of the fluorescence intensity for Dorsal staining in the entire cell as well as in the DAPI stained area of the cell for asrij null (I) and arf1 knockdown hemocytes (n = 10) (J). (K) Model indicating the suggested role of the ARF1-Asrij endocytic axis in regulating the Toll pathway. Error bars indicate standard error of mean. ** indicates P < 0.01 and *** indicates P-value < 0.001. Scale Bar: (G,H) 10 μm.