Fig. 6 | Nature Communications

Fig. 6

From: AWD regulates timed activation of BMP signaling in intestinal stem cells to maintain tissue homeostasis

Fig. 6

AWD/Tkv/MAD promote ISC quiescence and host resistance to acute infection. a Dynamic mitotic activity of ISCs when awd was specifically knocked down with two different RNAi lines was measured as numbers of phospho-Histone H3+ (pH3+) cells per gut during the course of an Ecc15 infection episode of 24 h. b Gut dysplasia was observed at posterior midgut 2 days after Ecc15 oral infection when tkv, mad, or awd were knocked down in ISCs respectively. Numbers of pH3+ cells per gut of these conditions were quantified. c, d Quantification of ISC mitotic activity in response to Ecc15 infection, when awd was knocked in ISCs with and without co-overexpressing Tkv-GFP fusion protein (c) or constitutively active Tkv (TkvQD, d), measured as numbers of pH3+ cells per gut. e Portion of Smurf flies when tkv, mad or awd were knocked down in ISCs respectively, monitored after a prior feeding on PE for 48 h. f Portion of Smurf flies when awd was knocked down in ISCs and EBs (with esgG4, tubG80ts driver) with and without TkvQD co-overexpression, monitored after a prior feeding on PE for 24 h. g Survival rates of flies (same as e) in response to acute intestinal damage were monitored after continuous PE infection. h Survival rates of flies (same as f) in response to acute intestinal damage were monitored after continuous PE infection. Error bars indicate SEM (a n = 10–17 flies, b n = 8–29 flies, c n = 9–15 flies, d n = 10–13 flies, e n = 45, 50, 45 flies for mCherryRNAi samples; n = 45, 50, 47 flies for tkvRNAi samples; n = 60, 50, 51 flies for madRNAi samples; n = 50, 50, 46 flies for awdRNAi-1 samples, f n = 89, 86, 88 flies for mCherryRNAi samples; n = 80, 73, 56 flies for awdRNAi-1 samples; n = 58, 61, 61 flies for awdRNAi-1; UAS::tkvQD samples). P values in g and h were calculated from log rank test (cohort sizes: g n = 40, 53, 42, 81 flies for mCherryRNAi, tkvRNAi, awdRNAi, madRNAi, respectively; h n = 90, 62, 90, 60 flies for mCherryRNAi, awdRNAi, UAS:TkvQD, awdRNAi;UAS:TkvQD, respectively). Other P values from Student’s t-test: ****P < 0.0001; ***P < 0.001; **P < 0.01; *P < 0.05, NS, not significant. One representative image from 4 to 7 flies tested in a single experiment was shown in b. Experiments were reproduced twice (ac, g), or three times (d, h)

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