Mating-induced increase in pMad expression was suppressed by intestinal RNAi ((function (or and BMP signaling (or RNAi powered by test with Holms correction was utilized for panel C. (neuroendocrine cells) experienced no effect on the mating-induced increase in GSC quantity. The number Isoimperatorin of germaria analyzed is definitely Isoimperatorin demonstrated inside the bars in panel C. For statistical analysis, a Wilcoxon rank sum test was utilized for panel C. *** 0.001. Level pub = 50 m in panel A and B. Underlying data can be found in S1 Data. GSC, germline stem cell; NPF, neuropeptide F.(TIF) pbio.2005004.s001.tif (6.8M) GUID:?F6F8F3DA-06E5-46F5-82AA-E271B00E5C2D S2 Fig: Manifestation pattern of Isoimperatorin driver. (ACC) Representative images of adult female brains and midguts immunostained with anti-NPF antibody (green) or anti-GFP antibody (green) and monoclonal nc82 (neuropil marker; magenta) or phalloidin (magenta). (A, B) driver expressed in the brain and VNC but not in the ovary. (C) RNAi driven by the driver did not reduce anti-NPF levels in the brain and VNC. Level pub = 50 m (mind and VNC) and 100 m (ovary). NPF, neuropeptide F; Tkg-GAL4, Tk-gut-GAL4; VNC, ventral nerve wire.(TIF) pbio.2005004.s002.tif (8.8M) GUID:?CA8F7D3A-52BD-4D83-BA48-D5E0B44D37EC S3 Fig: Manifestation pattern of several drivers. Representative images of adult female brains and midguts immunostained with anti-GFP antibody (green), anti-NPF antibody (magenta), or phalloidin (magenta). These drivers were indicated in NPF-positive EECs. Anti-GFP signals were also recognized in the brain, VNC, and oviduct, but not the ovary. Level pub = 50 m (mind and VNC) and 100 m (ovary and midgut). AG, accessory gland; EEC, enteroendocrine cell; NPF, neuropeptide F; OV, oviduct; SP, spermatheca; VNC, ventral nerve wire.(TIF) pbio.2005004.s003.tif (7.2M) GUID:?4F3B5B28-2EA9-429D-B9A4-24BA26425F87 S4 Fig: Gut NPF is not involved in gut remodeling. (A, B) The number of mitotic cells (panel A) or size (panel B) of the posterior midgut in virgin or mated woman flies was not affected by RNAi driven by (NPF-positive EECs). (C) Rate of recurrence of germaria comprising 1, 2, and 3 GSCs (remaining axis) and the average quantity of GSCs per germarium (ideal axis) in virgin (v) and mated (m) female flies. RNAi or RNAi driven by (ECs) or (NPF/Tk/Dh31-positive EECs) experienced no effect on the mating-induced increase in GSC quantity. Dots represent the number of mitotic cells in one middle midgut (panel A) or the diameter of a single posterior midgut (panel B); lines represent the median, and whiskers represent the interquartile range. For statistical analysis, a Wilcoxon rank sum test was used in panel A and C. Student test was used in panel B. *** 0.001 and ** 0.01. Underlying data can be found in S1 Data. Dh31, diuretic hormone 31; EC, enterocyte; EEC, enteroendocrine cell; gce, germ cell-expressed bHLH-PAS; GSC, germline stem cell; Met, Methoprene tolerant; NPF, neuropeptide F; pH3, phospho-histone H3; Tk, Tachykinin; Tkg-GAL4, Tk-gut-GAL4.(TIF) pbio.2005004.s004.tif (719K) GUID:?4926365E-407D-4F21-8D2C-B0A2504FBF48 S5 Fig: SP signaling controls NPF accumulation in midgut EECs. (A, C) Representative images of anti-NPF antibody immunostaining Isoimperatorin in the middle midgut LAMP3 are demonstrated on the remaining. Quantification of anti-NPF transmission intensity in the middle midgut is demonstrated on the right graph. Anti-NPF transmission intensity did not switch after overexpressing (mRNA level did not change in animals. (C) NPF build up was reduced by silencing in the middle midgut did not switch by this manipulation. Dots Isoimperatorin symbolize the relative transmission intensity of anti-NPF in one middle midgut (panel A and C) or relative expression levels of in the middle midgut (panel B and D); lines represent the median, and whiskers represent the interquartile range. For statistical analysis, a Wilcoxon rank sum test with Holms correction was utilized for panel A and C. College student test with Holms correction was utilized for panel B and D. *** 0.001 and ** 0.01; NS, nonsignificant ( 0.05). Level pub = 50 m in panel A and C. Underlying.
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