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. Author manuscript; available in PMC: 2023 Nov 3.
Published in final edited form as: Nature. 2023 May 3;617(7962):798–806. doi: 10.1038/s41586-023-06039-y

Extended Data Figure 1. Related to Figure 1. Characterization of Pi starvation and PXo knockdown.

Extended Data Figure 1.

a, b, pH3 staining of midguts fed 7d normal or PFA food. c, d, pH3 staining of midguts fed 9d CDF containing standard (100%) or 10% Pi. e, RT-qPCR measurement of MFS10 knockdown in the midgut when RNAi (VDRC line) is expressed 5d ubiquitously. The representative results with N=3 technical replicates are presented. p=0.0217. f, g, Midguts with 2d induced H2B-RFP expression in ECs, followed by 9d on normal or PFA food. h, i, j, k, Apoptag staining of midguts expressing Luc RNAi or PXo RNAi in ECs for 7d, with normal food (h, k), PFA, or the last 2d on Bleo. l, m, pH3 staining of midguts expressing Luc RNAi or PXo RNAi in ECs for 7d. n, o, pH3 staining and r, mitosis quantification of midguts with PXo RNAi (VDRC line) expression in ECs for 7d. N=8 (Ctrlw), 9 (PXo-iv). P=0.0193. p, q, pH3 staining of midguts expressing Luc RNAi or PXo RNAi in progenitors for 8d. s, Mitosis of midguts with PXo RNAi (NIG) expression in ECs for 9d. N=12 (Ctrlw), 15 (PXo-iN). P=7.94E-6. t, Mitosis of midguts expressing Luc RNAi or PXo RNAi alone, or together with p35 in progenitors for 9d, with or without the last 2d on Bleo. N=14 (Luc-i Normal, Luc-i p35, PXo-i p35), 13 (PXo-i Normal, Luc-i Bleo, PXo-i Bleo). P values from left to right: 9.97E-8, 3.00E-3, 1.92E-7. u, Mitosis of midguts with or without PXo RNAi (NIG) expression in progenitors for 8d, fed normal food (P=5.78E-5) or 2d Bleo (P=0.0237). From left to right: N=14, 13, 8, 12. v, Mitosis of midguts expressing GFP or p35 alone, or together with PXo RNAi in ECs for 9d. From left to right: N=12, 10, 12, 14; P=3.09E-6, 5.05E-3, 2.07E-7. w, Mitosis of midguts expressing Luc RNAi or PXo RNAi in progenitors, and fed normal, PFA, or Pi food for 7d. From left to right: N=8, 9, 8, 7, 8, 9; P=0.0132, 1.00, 1.40E-3, 1.18E-3, 1.00. x, Mitosis of midguts with or without PXo-HA expression in ECs for 8d, fed normal food or 2d Bleo. From left to right: N=12, 12, 13, 13; P=3.85E-7, 0.456, 3.85E-7. Data are mean ± SEM. P values are from two-tailed Welch’s t test (e), two-tailed Mann-Whitney u test (r, s, t, u, v, x, between Luc-i and PXo-i Normal in w) or one-way ANOVA with Bonferroni’s multiple comparison test (w). Arrowheads highlight pH3+ examples (a-d, l-q). Scale bars, 50 μm.