Journal: bioRxiv

Article Title: FGFR1 inhibition improves therapy efficacy and prevents metabolic adaptation associated with temozolomide resistance in glioblastoma

doi: 10.1101/2025.01.11.632515

Figure Lengend Snippet: (A) Heatmap of gene expression fold change from RNA-seq analysis in CCF cells treated with TMZ (100 µM) and FGFR1 (PD166866, 5 µM). (B, C) Volcano plot from RNA-seq analysis for upregulated (red) and downregulated (blue) genes for the indicated conditions. (D, E) Gene Ontology (GO) enrichment analysis for biological processes for the up-regulated (D) and down-regulated (E) genes for the dual treatment TMZ plus FGFR1i compared to DMSO (control). (F) GSEA plots for cell cycle related hallmarks comparing gene sets from TMZ treatment and dual treatment with TMZ and FGFR1i. (G) Volcano plot for upregulated (red) and downregulated (blue) metabolism-related genes. (H) GO plot representing the metabolic pathways with dysregulated genes by the treatment with TMZ and FGFR1i. (I-L) mRNA expression levels assessed by qPCR for genes found dysregulated in the RNA-seq analysis and highlighted in (G).

Article Snippet: Gene Set Enrichment Analysis (GSEA) was analysed using the GSEA v2.0.14 software (GSEA, Broad Institute, Cambridge, MA, USA) for Linux (v4.3.2) with 1000 permutations [ ].

Techniques: Expressing, RNA Sequencing Assay, Control

Journal: Scientific Reports

Article Title: Spautin-1 inhibits mitochondrial complex I and leads to suppression of the unfolded protein response and cell survival during glucose starvation

doi: 10.1038/s41598-022-15673-x

Figure Lengend Snippet: Autophagy inhibitors fail to show preferential cytotoxicity and suppress induction of ATF4 and XBP1s in glucose-starved or 2DG-stressed HT1080 cells. ( a ) HT1080 cells were treated with spautin-1 (10 μM) or SAR405 (1 μM) in HBSS containing HCQ (30 μM) for 4 h. Autophagosomes were visualized with the CYTO-ID Autophagy detection kit 2.0. ( b ) Effects of bafilomycin A1 (Baf), HCQ, and SAR405 on cell viability in GS-stressed HT1080 cells were determined by the CellTiter-Glo luminescent cell viability assay. Data are shown as mean ± SD ( n = 3). ( c ) Effects of spautin-1 (10 μM) and other autophagy inhibitors (Baf; 10 nM, HCQ; 30 μM, SAR405; 10 μM) on nuclear ATF4 and XBP1s induction under 2DG-stressed conditions were visualized using the Operetta CLS. Blue, red, and green fluorescent signals indicate nuclei, ATF4, and XBP1s, respectively. ( d ) Mean intensities of nuclear ATF4 and XBP1s intensities were determined using Harmony high-content analysis software. Data are shown as mean ± SD ( n = 3).

Article Snippet: Fluorescent images (nine fields per well) were acquired using a 20 × water objective lens by Operetta CLS (Perkin Elmer).

Techniques: Cell Viability Assay, High Content Screening, Software

Journal: Scientific Reports

Article Title: Spautin-1 inhibits mitochondrial complex I and leads to suppression of the unfolded protein response and cell survival during glucose starvation

doi: 10.1038/s41598-022-15673-x

Figure Lengend Snippet: USP10 and USP13 silencing has little effect on the UPR and cell viability under GS- or 2DG-stressed conditions. ( a ) Effects of USP10 and USP13 silencing on GRP78 in HT1080 cells were determined by western blotting. RPS3 was used as a loading control. The blot membranes were cut prior to hybridization with antibodies, according to Full range rainbow molecular weight markers. Original blots were presented in Supplementary Fig. . ( b ) Effects of USP10 and USP13 silencing on ATF4 and XBP1s induction in vehicle- or spautin-1-treated HT1080 cells under 2DG-stressed conditions were visualized using the Operetta CLS. ( c , d ) Mean intensities of nuclear ( c ) ATF4 and ( d ) XBP1s in ( b ) were determined using Harmony high-content analysis software. Data are shown as mean ± SD ( n = 3). ( e ) Effects of USP10 and USP13 silencing on cell viability under GS were determined by the CellTiter-Glo luminescent cell viability assay. Data are shown as mean ± SD ( n = 3). ( f ) Effects of USP10 and USP13 silencing on preferential cytotoxicity of spautin-1 under GS were determined by the CellTiter-Glo luminescent cell viability assay. Data are shown as mean ± SD ( n = 3).

Article Snippet: Fluorescent images (nine fields per well) were acquired using a 20 × water objective lens by Operetta CLS (Perkin Elmer).

Techniques: Western Blot, Control, Hybridization, Molecular Weight, High Content Screening, Software, Cell Viability Assay