Journal: Nature Communications
Article Title: Asymmetric histone inheritance regulates olfactory stem cell fates during regeneration
doi: 10.1038/s41467-026-70987-y
Figure Lengend Snippet: A Scheme of paired-daughter cell preparation for scRNA-seq. B Live imaging of cultured individual HBC to track the division from a mother cell to two daughter cells. C Clustering of HBC paired cells (left panel) after integrating scRNA-seq data of the current study ( GSE286046 ) into published dataset ( GSE99251 and GSE95601 ). Solid lines indicate asymmetric pairs cross different clusters. Dotted lines indicate symmetric pairs in the same clusters. The right panel shows the clusters of OE cells derived from GSE99251 and GSE95601 . D Feature plots showing expression of HBC marker genes Trp63 , Krt14 and Krt5 in collected single cells. E Feature plots showing expression of cell cycle and wound response genes Mki67 , Rps6 and Krt6a of collected single cells. F Statistics of paired information showing symmetric and asymmetric divisions in ( C ): 31.3% (15/48) of HBC pairs show asymmetric divisions. G Expression of H3.3 transcripts ( H3f3a / H3f3b ) in an individual pair. Red lines indicate the asymmetric pairs ( N = 15). Grey lines indicate the symmetric pairs ( N = 33). The asymmetric cutoff for H3f3a and H3f3b is 1.5. ( H ) Expression patterns of differentiation-associated genes (e.g., Krt16 , Sox9 , Ascl3, and Il33 ) (top panel) and self-renewal-associated genes (e.g., Trp63 , Fzd7 , Krt14, and Krt5 ) (bottom panel) in pairs with asymmetric H3.3. I PCA plot using scRNA-seq data of all renewed and activated HBCs. J MA-plot shows differentially expressed genes between renewed and activated HBCs. 85 genes (labeled in red) show higher expression in activated HBCs, 10 genes (labeled in blue) show lower expression in activated HBCs. K Differential expression of Mki67 , Suv39h1 , Krt16, and Krt23 in activated and renewed HBC cells. Note that Krt16 and Krt23 are identified by the current analysis. Gene counts normalized by external ERCC spike-in are used in the violin and box plots. All box plots include the median and data between the 25th and 75th percentile, with the whiskers indicating lower and upper extremes after removing outliers. The black ‘x’ within each box shows the average within each group. A two-tailed t-test was used to calculate P -values. L Asymmetric (31%) and symmetric (69%) SUV39H1 distribution in p63+ telophase HBCs ( N = 29) from culture. p63 shows the same distribution pattern as SUV39H1. Scale bar: 5 µm in ( L ). The cartoon elements are created in BioRender. Chen, X. (2026) https://BioRender.com/yh44iq5 and Adobe Illustrator. Source data are provided as a Source Data file.
Article Snippet: The primary antibodies and dilutions used were as follows: mouse monoclonal anti-p63, 1:100 (D-9, sc-25268; Santa Cruz); rabbit monoclonal anti-Ki67, 1:250 (NB600-1252, NOVUS); rabbit polyclonal anti-RNA polymerase II RPB1 phospho S2, 1:800 (AB5095, Abcam); rabbit polyclonal anti-RNA polymerase II RPB1 phospho S5, 1:800 (AB5131, Abcam); rat monoclonal anti-RNA polymerase II CTD, 1:200 (61082, Active Motif); rabbit polyclonal anti-Histone H4, 1:200 (Ab10158, Abcam), rabbit monoclonal anti-Histone H3, 1:200 (A17562, ABclonal); rabbit monoclonal anti-Histone H3.3, 1:250 (NBP2-67530, NOVUS); ChromoTek Histone-Label Atto488 (for H2A-H2B), 1:400 (tba488, Proteintech); rabbit monoclonal anti-phospho-Histone H3Thr3 (JY325), 1:800 (05-746 R, Millipore Sigma); Goat anti-Olfactory Marker Protein (OMP), 1:500 (544-10001, WAKO).
Techniques: Imaging, Cell Culture, Derivative Assay, Expressing, Marker, Labeling, Quantitative Proteomics, Two Tailed Test
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