Figure 3 from Functional RNAi Screening Identifies G2/M and Kinetochore Components as Modulators of TNFα/NF-κB Prosurvival Signaling in Head and Neck Squamous Cell Carcinoma
Year: 2024
Type: other
Abstract: Knockdown of G2/M kinases and kinetochore proteins inhibits NF-κB activity and sensitizes HNSCC cells to TNF-induced cell death. A, Quantification of percentage nuclear RELA in UMSCC1 cells transfected with siRNA targeting PLK1, AURKA, TPR, NUF2, and NDC80 for 72 hours, with TNFα (20 ng/mL) added for the final 30 minutes. DAPI (4′, 6-diamidino-2-phenylindole) was used as a nuclear counterstain. Scale bar, 10 μmol/L. Data represent the percentage nuclear localization of RELA from 15 cells and were analyzed using ImageJ as previously (12). B, Representative immunofluorescence images of the data in A. C, Colony formation assay of UMSCC1 cells transfected with siRNA targeting PLK1, AURKA, TPR, NUF2, and NDC80 for 72 hours, with TNFα (20 ng/mL) added for the final 24 hours. DAPI was used as a nuclear counterstain. D, Cell-cycle analysis of UMSCC1 cells transfected with siRNA targeting PLK1, AURKA, TPR, NUF2, and NDC80 for 72 hours, with TNFα (20 ng/mL) added for the final 24 hours. Statistics are shown in Supplementary Table S5. E, Annexin V analysis of UMSCC1 cells were transfected with siRNA targeting PLK1, AURKA, TPR, NUF2, and NDC80 for 72 hours, with TNFα (20 ng/mL) added for the final 24 hours. Bars represent the means ± SD. All experiments are representative of at least three biological replicates. NS, not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001.
Authors: Ethan L. Morgan, Anthony D. Saleh, Shaleeka Cornelius, Sophie G. Carlson, Tiffany Toni, Hui Cheng, Jun Jeon, Ramya Viswanathan, Xinping Yang, Christopher Silvin, Paul E. Clavijo, Anastasia L. Sowers, James B. Mitchell, Pinar Ormanoglu, Madhu Lal Nag, Scott E. Martin, Zhong Chen, Carter Van Waes
Language: English
Topic: NF-κB Signaling Pathways
Subfield: Cancer Research
Domain: Life Sciences
Open Access status: gold
OpenAlex ID: W4404147351