Fig. 5: Dynamics of immune cell infiltration over time.
From: Single-cell RNA sequencing reveals evolution of immune landscape during glioblastoma progression
a, UMAPs of normal brain, Early and Late stage GBMs. b,c, Relative number of EGFR+ (b) and the indicated cell types (c) from scRNA-seq data and flow cytometry of the GBM samples used for scRNA-seq. d, Cluster composition of microglia and macrophages from scRNA-seq. e, Flow cytometry of EGFR+ and CD45+ cells from GBMs. Data are mean ± s.e.m. of biologically independent replicates. Control brain day 0 n = 3, GBM n = 6, 6 and 3 for days 10–15, 20–25 and >30, respectively. At >30 days relative to day 0, *P = 0.0001, **P = 0.0285, unpaired t test two-tailed. f, Longitudinal flow cytometry of the indicated GBM cell types. Data are mean ± s.e.m. of biologically independent replicates. Left panel, control brain day 0 n = 3, GBM n = 8, 8 and 9 for days 10–15, 20–25 and >30, respectively, *P = 0.0122, **P = 0.0236, ***P = 0.0488, ****P = 0.0006, *****P = 0.0079, unpaired t test two-tailed. Right panel, day 0 n = 3, GBM n = 7, 5 and 3 for days 10–15, 20–25 and >30, respectively. At >30 days relative to day 0, all P > 0.05 and considered not significant, unpaired t test two-tailed. g,h, Pseudotime trajectory (seed cluster 12) (g) and DEGs (h) of oligodendrocyte clusters 9 and 12. i,j, Assessment of GBM BBB integrity. Representative photomicrographs of GBM brains post EB and NaF administration and quantification (i). Data are mean ± s.e.m. of biologically independent replicates. EB, n = 3, 5, 4, 5 and 4, for days 0, 10, 20, 25 and 30+, respectively and NaF, n = 2, 5, 4, and 3 for days 0, 10, 20, and 30+, respectively. *P = 0.0209, **P = 0.0012, ***P = 0.0089 when compared with day 0, unpaired t test two-tailed. Representative gadolinium-enhanced T1-weighted MRIs and quantification of a developing GBM imaged at 22, 26 and 32 days post-tumor initiation (j). Tumor volumes (in mm3) were measured using T2 weighted images. Scale bar, 2 mm; data represent mean ± s.d. of technical replicates. *P < 0.0001, **P = 0.0071, ***P = 0.0029, ****P = 0.0002 when compared with 22-day voxel intensities, unpaired t test two-tailed.
