Cell annotation, trajectory analysis and GSEA of two GBM cell subsets with distinct differentiation patterns. (A) All 13 clusters of cells in GBMs were annotated by singleR and CellMarker according to the composition of the marker genes. (B) Trajectory analysis revealed two subsets of GBM cells with distinct differentiation patterns. GBM CSCs were mainly located in the root, whereas GBM cells were located in either branch. Branch I GBM cells were defined by the type I GBM cell subset (434 GBM cells). Branch II GBM cells were defined by the type II GBM cell subset (444 GBM cells). (C and D) GSEA of type I and II GBM cell subsets was performed to identify related molecular mechanisms and pathways. An FDR ≤ 0.05 was considered statistically significant.