Figure 2 from Nuclear Focal Adhesion Kinase Protects against Cisplatin Stress in Ovarian Carcinoma
Year: 2024
Type: other
Abstract: Nuclear FAK accumulation occurs upon cisplatin treatment of human and murine ovarian tumor cells. A, Nuclear and cytoplasmic cell fractionation of human OVCAR4 cells treated with DMSO (control) or cisplatin (1 μmol/L) for 12 hours followed by immunoblotting for FAK pY397, total FAK, β-tubulin (cytoplasmic marker), and histone H3 (nuclear marker). B, Schematic of GFP fusion to FAK denoting N-terminal band 4.1, ezrin, radixin, moesin homology (FERM), kinase, and focal adhesion targeting (FAT) domains. FAK FERM arginine R177 and R178 to alanine mutational changes prevent FAK nuclear localization. C, Stable reexpression of GFP-FAK-WT (WT) and GFP-FAK-NLS− (NLS−) in KMF FAK KO murine ovarian tumor cells as analyzed by FAK, pY397 FAK, and β-actin immunoblotting. D, KMF FAK-WT and KMF FAK-NLS− cells treated with DMSO (control) or cisplatin (20 μmol/L) for 1 hour and immunoblotting for FAK pY397 and total FAK. E, Confocal immunofluorescence imaging was used to visualize GFP-FAK and DAPI stain in FAK-WT and FAK-NLS− cells treated with DMSO (control) or cisplatin (20 μmol/L) for 12 hours. Shown are representative images of GFP-FAK (images shown at midline of Z-stack), nucleus, and merged images. Scale bar, 10 μm. F, Nuclear and cytoplasmic fractionation of FAK-WT and FAK-NLS− cells treated with cisplatin (20 μmol/L) for 0, 12, or 24 hours followed by immunoblotting for FAK, β-tubulin, and histone H3. G, Representative images of GFP-FAK-WT and FAK-NLS− expressing (images shown at midline of Z-stack) and DAPI-stained cells treated with leptomycin B (50 nmol/L) for 6 hours. Scale bar, 10 μm. Cyto, cytoplasmic.
Language: English
Subfield: Reproductive Medicine
Field: Medicine
Domain: Health Sciences
Open Access status: gold
OpenAlex ID: W4405648186