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. Author manuscript; available in PMC: 2021 Jun 23.
Published in final edited form as: Cell Rep. 2021 May 18;35(7):109154. doi: 10.1016/j.celrep.2021.109154

Figure 5. G-quadruplex-binding ligands re-suppress activated hTERT in glioblastoma multiforme with mutations in the hTERT promoter.

Figure 5

(A and B) hTERT expression (A) and telomerase activity (B) in glioblastoma multiforme (GBM) cell lines U87MG and LN229 (with −124G > A hTERT promoter mutation) following treatment with G-quadruplex-binding ligands SMH1−4.6 and JD83 (2.5 μM) or DMSO for 24 h.

(C and D) TRF2 ChIP-qPCR spanning the hTERT promoter following treatment with SMH1−4.6 and JD83 (2.5 μM) or DMSO for 24 h in LN229 (C) or U87MG (D) cells.

(E and F) Fold change in repressor histone mark H3K27me3 by ChIP-qPCR spanning the hTERT promoter following treatment with SMH1−4.6 and JD83 (2.5 μM) or DMSO for 24 h in LN229 (E) or U87MG (F) cells. Fold-change shown with respect to total H3 ChIP; respective ChIPs were normalized to 1% input.

(G) TRF2 ChIP-qPCR at the exogenously inserted CCR5-locus-hTERT promoter with either the wild-type or −124G/−146G > A hTERT promoter mutations in HEK293T cells following treatment with SMH1−4.6 and JD83 (2.5 μM) or DMSO-treated (control) for 24 h. Normalized to IgG ChIP in each case.

(H) GABPA ChIP followed by qPCR at the hTERT core-promoter (+38 to −237 bp); in U87MG (−124G > A mutant) cells following treatment with SMH1−4.6 and JD83 (2.5 μM) or DMSO (control) for 24 h. Normalized to IgG in each case. TFB1M and TFB2M are positive control and B-actin is negative control for GABPA ChIP. All error bars represent ± SDs from mean values. p values calculated by paired/unpaired t test or two-way ANOVA (*p < 0.05, **p < 0.01, ***p < 0.005, ****p < 0.0001).