Extended Data Fig. 10: ASO-mediated inhibition of HELDR synergistically enhanced anti-GBM activity of EGFR inhibitor erlotinib.
From: An EGFR co-amplified lncRNA HELDR promotes glioblastoma malignancy through KAT7-driven gene programs
a, qRT–PCR analysis of GSC11 cells 24 h after transfection with 100 nM HELDR-targeting ASOs or a control ASO in vitro (n = 3). b, c, IB for indicated proteins in GBM6 and GSC11 cells 72 h after transfection with 100 nM HELDR-targeting ASOs or a control ASO in vitro. d, Cell viability of EGFR-amplified (GSC11, GBM6) and non-amplified (GSC1478, GSC157) GSC/GBM cells after 6 days of indicated ASO treatment (n = 3). e-j, Cell proliferation (n = 3) (e-g), and glioma sphere formation (h-j) with indicated treatment. In d-j, ASO, 100 nM; Erlotinib, 0.8 μM. k, Analysis of coding DEGs from RNA-seq of GSC17 cells 24 h after transfection with 100 nM control or HELDR-targeting ASO in vitro (two biological replicates; threshold: |fold change|> 1.5 and adjusted p < 0.05). Venn diagrams showing common coding DEGs between HELDR knockdown by shRNA (shared by both shRNAs; see Fig. 3n) and ASO treatment. l, BLI images for mice bearing GSC11 brain tumour xenografts with indicated treatments (n = 5 in each group). Treatments: ASO, 4 µg/mouse, twice/week, erlotinib, 50 mg/kg, 5 days on/2 days off; until morbibund. m, qRT–PCR analysis of HELDR expression in tumour area from brain sections (n = 4). n, Representative images of IF staining for brain sections with GBM6 xenograft tumours with indicated treatments. o-q, Quantification of IF staining shown in n (n = 5). r-t, Quantification of IF staining for brain sections with GSC11 xenograft tumours with indicated treatments (n = 5). Scale bar, 100 μm. β-actin is a sample processing control in b and c. All blots, qRT-PCR, and in vitro assays are representative of three independent experiments. One-way ANOVA with Dunnett’s post hoc test in a, two-sided unpaired t-test in d, one-way ANOVA with Tukey’s post hoc test in e-g, m, and o-t, two-sided likelihood ratio test in h-j, two-sided Wald test with Benjamini–Hochberg adjustment for multiple comparisons in k, Data are presented as mean ± s.d. (a, d, e-g, m, and o-t).
