Figure 5. LAM loss in fat bodies causes midgut hyperplasia and reduces survival.

A–J. Compared to control young midguts (A, B), midguts from young Lam mutant flies (E, F), flies with fat body-specific Lam depletion (G, H), or old control flies (C, D) exhibited hyperplasia. esg-GFP, green; pH3, red. Fat body-specific LAM expression in young Lam mutant flies significantly reduced hyperplasia (I, J). Images show the face on view (A, C, E, G and I) or cross-section view (B, D, F, H and J, revealing the basally localized esg-GFP+ ISCs and inappropriately differentiated EBs) of midguts. Arrows and arrowheads indicate the luminal and muscular surfaces, respectively, as revealed by phalloidin staining of actin (red). DAPI (blue), nuclei. All flies were kept in conventional condition. Scale bars, 20 μm.

K. Midgut hyperplasia based on esg-GFP. The percent of esg-GFP+ cells (% intestinal hyperplasia) was grouped into four classes and plotted. *p<0.05, **p<0.01, ***p<0.001, Wilcoxon two-sample test.

L. Midgut cell proliferation based on pH3 staining. The average number of pH3+ cells was counted from the whole midgut and plotted. Error bars, SEM. Student’s t-tests, **p<0.01, ***p<0.001.

M. Survival rate of Lam-mutant and flies depleted of LAM in fat body. Error bars, SEM based on three independent experiments performed with 200 larvae per experiment. Student’s t-tests, **p<0.05, **p<0.01, ***p<0.001.

Con or Axe, conventional or axenic conditions. n, numbers of midguts analyzed.

See also Figure S5.