Supplementary Figure S8 from Heterogeneity in NECTIN4 Expression Across Molecular Subtypes of Urothelial Cancer Mediates Sensitivity to Enfortumab Vedotin
Year: 2023
Type: supplementary-materials
Abstract: (A) Validation of knockdown by cell surface staining of NECTIN4 protein expression in HT-1376 cells using CRISPRi/dCas9 and five unique sgRNAs targeting different regions of NECTIN4 gene and promoter. The sgNECTIN4-a guide was used for subsequent experiments. (B) Confirmation of sgNECTIN4 CRISPRi/dCas9-mediated NECTIN4 mRNA knockdown in HT-1376 cells by quantitative PCR, **p<0.01. All sgRNAs decreased NECTIN4 mRNA expression by >90%. (C) Phase-contrast, brightfield microscopy of HT-1376 cells in Control and NECTIN4 knockdown cells to show the morphology of cells. Scale bar = 400 mm. (D) Validation of knockdown by cell surface staining of NECTIN4 protein expression in HT-1197 cells using CRISPRi/dCas9. (E) EV dose-response WST proliferation assay in HT-1197 Control (black) and NECTIN4 (KD) knockdown (purple) cells.
Language: English
Subfield: Surgery
Field: Medicine
Domain: Health Sciences
Open Access status: gold
OpenAlex ID: W4361890315