Figure 3.

THIO induced DNA damage in glioma cells. (A-C) The correlation of IC₅₀ of THIO and the enrichment scores of telomere/telomerase-related gene signatures including "Telomere Maintenance," "Telomere Pathway" and "TERT Targets" in 11 glioma cell lines in CCLE. (D) Baseline telomerase activity of THIO sensitive cell lines and THIO resistant cell lines. (E-F) Relative expression of hTERT of M059K and DBTRG-05MG cells following treatment with THIO at 5 μM for 72 hours. (G) Western blot was performed to detect the protein expression of γH2AX in 8 glioma cell lines treated with THIO for 72 hours. (H) The correlation of IC₅₀ of THIO and the fold change of γH2AX expression. (I) Immunofluorescence staining of γH2AX in M059K cells treated with THIO at 0, 1, 2.5 and 5 μM for 48 hours. Blue indicated DAPI and green indicated γH2AX. (J) Quantification of immunofluorescence staining of γH2AX for H4 cell lines treated with THIO at 0, 1, 2.5 and 5 μM for 72h. (K) Immunofluorescence staining of γH2AX for 3 PDX tumor models including D09-0500MG, D09-0394MG, and D10-0021MG. Tumor-bearing mice was treated with control or THIO at 2.5 mg/kg every other day. Blue indicated DAPI and green indicated γH2AX. (L) Quantification of immunofluorescence staining of γH2AX for 3 PDX tumor models.

(M) Telomere dysfunction induced foci (TIF) assay for the D2363PXA cell lines. D2363PXA cells were treated with THIO at 1.5 uM for 48h following by fixed and performed TIF analysis. (N) Quantification of number of TIF per cell.