Journal: Communications Biology
Article Title: Stem cells resume asymmetric division upon niche re-entry through reactivating the centrosome orientation checkpoint
doi: 10.1038/s42003-026-09812-7
Figure Lengend Snippet: A The table shows the division orientation of dedifferentiated cells after reattachment to the hub in the steady-state conditions (upper table) (listed in Table ) and during the recovery period (lower table) after partial depletion of GSCs (listed in Table ). Dedifferentiated cells that did not divide after reattachment are not shown. Orientation was judged as “oriented” when one of the two spindle poles was closely associated with the hub during mitosis. B Schematic of centrosome orientation checkpoint (COC). When centrosomes are positioned perpendicularly towards the hub-GSC interface, COC is cleared and GSCs are allowed to enter mitosis. When centrosomes are mispositioned, COC blocks GSCs from progression to mitotic entry and GSCs are arrested in the late G2 phase. C , D Representative immunofluorescence images of Wild type ( yw , 6 x heat-shock treated and recovered for 3 days) and hs-bam (6 x heat-shock treated and recovered for 3 days) following 4.5 h of ex-vivo colcemid treatment. The testes were stained with phospho-histone-3 (PH3, red), Vasa (blue), FasIII (green), and Hts (green) antibodies. CySC=cyst stem cells. E , F Box plots show 25–75% (box), minimum to maximum (whiskers) with all data points. In E , each data point indicates percentage of the PH3 positive cells in scored testis after 4.5-h colcemid treatment. “n” indicates the number of scored testes. In F , changes in GSC number after forced differentiation of GSCs are shown. From pre-heat shock (pre-hs), post-heat shock (post-hs), to 3- and 5-day recovery points (3 d rec, 5 d rec) without or with expression of Baz RNAi are shown. nosGal4 driver was used to express RNAi construct against Baz. “n” indicates the number of scored testes. Šídák’s multiple tests calculated p -values in E , F . G A representative immunofluorescence image of a Baz-GFP Flytrap testis with centrosome staining (anti-γTub, or GTU-88). FasIII and γTub (red), Vasa (blue), Baz-GFP (green). Asterisks in all images indicate the approximate location of the hub. All scale bars represent 10 μm.
Article Snippet: The primary antibodies used were as follows: rat anti-Vasa (1:20; developed by A. Spradling and D. Williams, obtained from Developmental Studies Hybridoma Bank (DSHB); mouse anti-hu-li tai shao (Hts) (1:20, 1B1; DSHB); mouse-anti-Fasciclin III (FasIII) (1:40, 7G10; DSHB); mouse anti-γ-Tubulin (1:400, GTU-88; Sigma-Aldrich); rabbit anti-phosphorylated (Thr3) histone H3 (1:200; clone JY325, Upstate).
Techniques: Immunofluorescence, Ex Vivo, Staining, Expressing, Construct
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