Extended Data Fig. 8: Proliferation capacity, location and the lineage-related gene expression patterns of grafted cells.
From: Astrocyte specification in the mouse septum is shaped by both developmental origin and local signals
a) Heatmap analysis of snRNAseq data indicating cell cycle genes enriched in the first few days after birth, including Mki67, Top2a and Cdk1. b) Location of Zic4-transplanted astrocytes in the septum at P30. Different colors indicate different samples. The four grey dots in each plot represent septal region indicators: the left grey dot indicates the left wall edge of the septum, the right grey dot indicates the right wall edge of the septum, the grey dot in the MS indicates the center of the MS, the top grey dot marks the midline. These grey dots were averaged from different samples used for analysis. N = 7-10 transplants, a total of 39-40 cells were analyzed. c, d) Immunostaining for tdTomato, Nkx2.1 and Sox9 in homotopic (c) and heterotopic (d) Zic4-transplanted samples 5 days post-transplantation. White boxes indicate high magnification images shown in the right panels. White arrows indicate transplanted cells positive for tdTomato and Sox9, but negative for Nkx2.1. Scale bar: 100 µm (left), 50 µm (right). e) Quantification of the percentage of Nkx2.1-postive transplanted astrocytes in the entire astrocyte population 5-day post-transplantation, LS to MS (9.44%) and LS to LS (9.22%), N = 4 transplants per each condition. f, g) Immunostaining for tdTomato, Ki67 and Zic in homotopic (f) and heterotopic (g) Zic4-transplanted samples 5 days post-transplantation. White boxes indicate high magnification images shown in the right panels. White arrows indicate transplanted cells positive for tdTomato, but negative for both Ki67 and Zic. Scale bar: 100 µm (left), 50 µm (right). h) Quantification of Zic-positive transplanted cells in the transplanted population 5-day post-transplantation, LS to MS (40.9%) and LS to LS (37.5%), N = 4 transplants for each condition, i) Quantification of the percentage of Ki67-positive transplanted cells in the entire transplanted population, LS to MS (8.88%) and LS to LS (6.80%), N = 4 transplants. j) Representative images showing low Zic expression in transplanted cells in both conditions. White arrows indicate grafted cells with low Zic expression. Scale bar: 10 µm. Box plots in E, H and I show the median (center), 25th and 75th percentiles (box), and min to max (whiskers).
