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

. 2020 Jul 17;3:388. doi: 10.1038/s42003-020-1090-2

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

© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Fig. 6 — a Representative immunofluorescence images for eIF4A1 (red in merged imaged) and p62/SQSTM1 (green in merged imaged) in Atg12^f/f and Atg12^KO MEFs, nuclei were counterstained with Hoechst (blue). Yellow box indicates region of inset panel in the top left corner. Far right panels show the points of colocalization (white) of eIF4A1 in p62/SQSTM1. Bars = 50 μm. b Manders’ coefficient (percent of colocalization) of eIF4A1 in either NBR1 or p62/SQSTM1 in Atg12^f/f and Atg12^KO MEFs was calculated, and boxplot with dotplot overlay representing one field is shown (n = 3 biologically independent replicates). An outlier (Dixon test p = 0.03) was excluded from statistical analysis and the p values between Atg12^f/f and Atg12^KO were calculated by t test. c Representative immunoprecipitation of eIF4A1 and immunoblot for the autophagy cargo receptors p62/SQSTM1 and NBR1. Arrow indicates p62/SQSTM1, asterisk indicates immunoglobulin heavy chain. Immunoprecipitation was performed with three biologically independent replicates. d Protein lysate from Atg12^f/f and Atg12^KO MEFs that were knocked down for p62/SQSTM1 or treated with nontargeting shRNA was captured by cap pulldown, and the ratio of eIF4A1 to eIF4G1 was quantified (mean ± SD, n = 4 biologically independent replicates). e Protein lysate from Atg12^f/f and Atg12^KO MEFs that were knocked down for p62/SQSTM1 or treated with nontargeting shRNA or HEK293Ts that overexpress p62ΔLIR or empty vector control was captured by cap pulldown, and immunoblotted as indicated. f–h Protein lysate from Atg12^f/f or Atg12^KO MEFs stably infected with shRNA to p62/SQSTM1 or nontargeting control was collected. Relative quantification from immunoblots for f p62/SQSTM and g BRCA2, normalized to loading control are shown in the boxplot with dotplot overlay per biological replicate. p values were calculated using ANOVA with Tukey’s post hoc test and significant values are reported. h The change in BRCA2 levels between Atg12^f/f and Atg12^KO in shNT-treated cells or shp62-treated cells is plotted, p = 0.76 between shNT and shp62 for ΔBRCA2, calculated by t test.