Transcriptional Profiling of VNLG-152R-Treated MDA-MB-231 Cells

MDA-MB-231 cells were treated with 10 μM VNLG-152R for 24 h in triplicates. Total RNA was isolated using RNAeasy Plus mini kit (Qiagen) following manufacturer’s instructions. The RNA preparation was quantified and assessed its quality using Agilent 2100 Bioanalyzer. A RIN value of 8 or above was used for all samples. The sequencing libraries were prepared with the NEB Ultra II Directional RNA library prep kit. Further, the libraries were evaluated for quantity and size distribution using Qubit and Agilent 2100 Bioanalyzer. Sequencing was carried out on an Illumina NovaSeq S2 PE100 bp lane (Maryland Genomics, Institute for Genome Sciences, University of Maryland Baltimore). As a norm, Phred quality score (Q score; to measure the quality of sequencing) more than 90% of the sequencing reads reached Q30 (99.9% base call accuracy). Differential Gene Expression and Gene Set Enrichment analyses (GSEA) were performed to identify canonical cellular pathways modulated by VNLG-152R as reported previously (28 (link)).