Fig. 4: TRIP13 expression regulates GBM cell sensitivity to Osimertinib.

A Integrating multiple layers of genomic data to identify TRIP13 as a potential regulator of intrinsic EGFR-TKI resistance in GBM. Venn diagrams showing the overlap of Osimertinib-resistance (Osim-R-Up/Dn) genes, Gefitinib-resistance (Gefi-R-Up/Dn) genes, GBM-dysregulated genes (GBM-Up/Dn), and EGFR co-expressed genes (GBM-EGFR-Pos/Neg-Cor). Heatmap visualization of six common overlapping genes. The color bar denotes the Log2 (Fold-Change (FC)) of differentially expressed genes in GSE172002, TCGA-GBMLGG rnaseq datasets and pearson correlation (r) of EGFR co-expressed genes in CPTAC-GBM TMT-based proteomics dataset. Barplot represents log10 (Intensity Score (IS)) of EGFR-binding partner identified in our previously reported CoIP-MS dataset. B LN-229 and LN-18 cells with or without TRIP13 overexpression were treated with Osimertinib (5 μM) for the indicated times, and cell growth was determined by CCK-8 cell survival assay. The results were shown as mean ± SD, n = 6; *p < 0.05. C SF-539 and U87MG cells with or without TRIP13 knockdown were treated with Osimertinib (5 μM) for the indicated times, and cell growth was determined by CCK-8 cell survival assay. The results was shown as mean ± SD, n = 6; *p < 0.05, **p < 0.01. D LN-229 cells with or without TRIP13 overexpression were vehicle-treated or treated with Osimertinib (5 μM) for 24 h. Cell lysates were analyzed by immunoblotting (IB) using the indicated antibodies. E U87MG cells with or without TRIP13 knockdown were vehicle-treated or treated with Osimertinib (5 μM) for 24 h. Cell lysates were analyzed by IB using the indicated antibodies. F LN-229 cells with or without TRIP13 overexpression were treated the same as in D, and cell morphology was examined by phase-contrast microscopy (scale bar = 20 μm). The numbers of vacuolated and non-vacuolated cells were counted manually, and the ratio of vacuolated cells was calculated and shown as mean ± SD, n = 6, **p < 0.01. G U87MG cells with or without TRIP13 knockdown were treated the same as in E, and cell morphology was examined by phase-contrast microscopy (scale bar = 20 μm). The numbers of vacuolated and non-vacuolated cells were counted manually, and the ratio of vacuolated cells was calculated and shown as mean ± SD. n = 6, **p < 0.01.Data represent three independent experiments with similar results.