Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2018 Sep 11;10(7):1045–1059. doi: 10.1080/19420862.2018.1502025

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 2018 Taylor & Francis Group, LLC

PMC Copyright notice

Figure 6. — Mitochondrial delocalization of endogenous p53 in U2OS cells without apparent effect on cell viability. U2OS cells transiently express MOM/V5-tagged p53 TAD Nbs. The MOM/V5-tagged Fsc Nb2 was used as negative control. (a) Representative epifluorescence images demonstrating the enrichment of MOM/V5-tagged Nbs at the mitochondria, which were labeled via Mitotracker. (b) Endogenous p53 occasionally adopts a mitochondrial-like pattern in the presence of MOM/V5-tagged p53 TAD Nb1 and MOM/V5-tagged p53 TAD Nb8, albeit with much lower efficiency than observed for overexpressed p53. A similar pattern however cannot be observed in the presence of MOM/V5-tagged Fsc Nb2, where p53 is predominantly located in the nucleus. Visualization of the nuclei was achieved with DAPI, whilst p53 and the MOM/V5-tagged Nbs were visualized with DO-1 and an anti-V5 antibody, respectively. An XTT cell viability assay was performed to evaluate whether mitochondrial delocalization of endogenous p53 impacts cell viability. The assay was performed in the absence (c) or presence of 100 µM etoposide (d). Two negative controls were implemented, for which cells were either transiently transfected with the MOM/V5-tagged Fsc Nb2 (C1) or were subjected to a mock-transfection (C2). The graph represents the mean net absorbance of the formazan dye at 475 nm. A significant change in cell viability is only observed for MOM/V5-tagged p53 TAD Nb43-expressing cells, which display a higher proliferation rate and thus cell viability (P < 0.05, one-way ANOVA Dunnett’s multiple comparison test). (c) When cells are additionally treated with 100 µM etoposide, there is no difference in cell viability between cells that express MOM/V5-tagged p53 TAD Nbs and cells that express the MOM/V5-tagged Fsc Nb2 (one-way ANOVA, Dunnett’s multiple comparison test) (d). The overall viability of U2OS cells significantly decreases in the presence of 100 µM etoposide (P-value = 0.0017, two-way ANOVA). Nanobody treatment also seems to significantly impact cell viability (P-value = 0.0039, two-way ANOVA), however this effect is merely caused by MOM/V5-tagged p53 TAD Nb43. The impact of etoposide treatment is not influenced by the presence of MOM/V5-tagged Nbs (P = 0.0505, two-way ANOVA) ((c) and (d)).