Supplementary Figures and tables from A Functional Screen Identifies miRs That Induce Radioresistance in Glioblastomas
Year: 2023
Type: supplementary-materials
Abstract: S1A. List of miRNAs enriched independent of IR. S1B. Clonogenic survival assay was performed with 2 Gy IR in 12 different GBM lines. Bar graphs were generated with Mean {plus minus} S.D. of 3 independent experiments. S2A. IR sensitivity of LN229 with individual stable expression of miR-1, miR-125a, miR-150 and miR-425 was determined by clonogenic assay. Curves were generated with Mean {plus minus} S.D. of 3 independent experiments. For statistical analysis the areas under the curves were calculated by approximating them to trapezoids. Comparison was done with one-way ANOVA (p < 0.05) and Tukey's post hoc (p < 0.05) test. S2B. Expression of fused miR-1, miR-125a, miR-150 and miR-425 from pcDNA3.1 vector. S3. A) Illustration shows the construct of GFP-fused miRNA sponges. GFP is fused at the 3' end of 4 identical repeats of miRNA sponges in tandem. The sequence of each miRNA sponge is shown below. (B) Inhibition of GFP expression fused to miRNA sponges was tested by transfecting corresponding miRNA mimics. Comparison was done with one-way ANOVA (p < 0.05) and Tukey's post hoc (p < 0.05) test. S4A. Impact of individual miR-scr, miR-1, miR-125a, miR-150 and miR-425 and combined miR-4x' on apoptosis in LN229 cells. Cells were exposed to 10 Gy γ-IR and the activation of Caspase 3 and 7 was determined with Caspase-Glo 3/7 Assay System. Comparison was done with one-way ANOVA (p < 0.05) and Tukey's post hoc (p < 0.05) test. S4B. Phosphorylation of Chk1 S345 in LN229 cells transfected with indicated miRNA upon exposure to 10 Gy IR. The mean {plus minus} S.D. of 3 independent experiments is demonstrated. Comparison was done with one-way ANOVA (p < 0.05) and Tukey's post hoc (p <0.05) test. S5. Expression of PAI-1 was examined upon stimulation with TGF-β with or without TGF-β inhibitor in indicated GBM cell lines. Bar graphs were generated with Mean {plus minus} S.D. of 3 independent experiments. Comparison was done with one-way ANOVA (p < 0.05) and Tukey's post hoc (p < 0.05) test. Supplementary Table 1. Transcription factors that are predicted by prediction algorithms, MotifScanner and CisGenome, to regulate the expression of all 4 miRNAs within 5000 base pairs upstream of each miRNA. The commons factors predicted for all 4 miRNAs were listed in the table. Supplementary Table 2. Selected genes from our predictions that overlap with the list of factors whose silencing cause radioresistance have been shown in Table 2.
Language: English
Subfield: Cancer Research
Domain: Life Sciences
Open Access status: gold
OpenAlex ID: W4362486496