Fig. 6

Tumor-specific, radiosensitizing function of lncGRS-1 knockdown in mature brain organoids (MBOs). a Schematic of MBO assembly from induced mature astrocytes (iAstrocytes) and induced neurons (i³Neurons). b Single molecule RNA FISH of lncGRS-1 in iAstrocyte MBO (A-MBO) cells following transfection of non-targeting ASO (top) or ASO targeting lncGRS-1 (bottom). Scale bar = 5 μm. c RT-qPCR of lncGRS-1 in A-MBOs seeded with DIPG SF8628 following ASO transfection (n = 2 biological replicates per condition; error bar = SD). d Left, fluorescence viability assay of A-MBOs following transfection of non-targeting ASO or ASO targeting lncGRS-1. Right, apoptosis induction assay of A-MBOs following transfection of ASOs (n = 2 biological replicates per condition; error bar = SD). e Schematic of RFP+ glioma cell seeding and subsequent RFP+ tumor growth over time. f, g Longitudinal fluorescence microscopy of A-MBOs seeded with RFP+ DIPG SF8628 cells treated with non-targeting Ctrl ASO (f) or ASO targeting lncGRS-1 (g). h, i Quantification of RFP+ tumor burden (h) and organoid diameter (i) in longitudinal analysis of A-MBOs seeded with RFP+ DIPG SF8628 cells (n = 6 biological replicates per condition; two-tailed Student’s t test). j Quantification of RFP+ U87 GBM tumor burden in iAstrocyte and i³Neuron MBOs (AN-MBOs) treated with ASOs with or without radiation (n = 5 biological replicates per condition; two-tailed Student’s t test). Boxplots represent 1st quartile, median, and 3rd quartile with whiskers = range