Fig. 3

iGRPs form LGGs in Rag1^−/− mice a Differentiation of human iNPCs into iOPCs, iGRPs and astrocytes. b Rag1^−/− mice harbor LGGs at 1mpi following CTL, 2041C > T^−/− and 6513T > A^−/− NF1-null iGRP and iOPC brainstem injections, as well as KIAA1549:BRAF-expressing iGRP and iOPC cerebellum injections. No LGGs were observed following NF1-null or KIAA1549:BRAF-expressing hiPSC-astrocyte injections. G, glial only; G/E, mixed embryonal/glial. c Analysis of 2041C > T^−/− NF1-null differentiated cells revealed CD133⁺, SOX2⁺, ABCG1⁺, O4⁺, GFAP⁺ and S100β ⁺ iGRPs (top panel), GFAP⁺, S100b⁺, EAAT⁺ and EAAT2+ astrocytes (middle panel) and O4⁺, MBP⁺, GFAP⁺ and NG2^neg iOPCs ( lower panel). d Representative low-magnification H&E images of 2041C > T^−/− and 6513 T > A^−/− NF1-null and KIAA1549:BRAF-expressing iGRP and iOPC LGGs. Scale bars, 1 mm. e H&E, Ku80, Ki67, GFAP, OLIG2, synaptophysin, BLBP, CD133, ABCG1, PDPN, SOX10 and P16 immunostaining images of representative 2041C > T^−/− NF1-null iGRP- (top) and iOPC- (bottom) LGGs at 1mpi. Scale bars, 100 µm. (f) Summary of relative immunopositivity scoring for GFAP, OLIG2, synaptophysin and Ki67 in LGG-bearing 1mo mice. −, 0%; +, < 25%; ++, 50%; +++, > 75% immunopositive cells. Data are represented as means ± SEM, one-way ANOVA with Bonferroni post-test correction, p values are not significant