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

. 2019 Nov 19;19(1):368–378. doi: 10.3892/ol.2019.11116

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

Copyright: © Li et al.

This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.

PMC Copyright notice

Figure 3. — Poly (ADP-ribosylation) of HMGB1 facilitates its acetylation. (A) Cell lysates were immunoprecipitated with an HMGB1 antibody, followed by western blot analysis. The acetylation or poly (ADP-ribosylation) levels were measured using antibodies specific for acetylated lysine or poly (ADP-ribosylation). β-actin was used as a loading control. Quantified data are presented (HMGB1/β-actin, PARylation-HMGB1 or Acetylated-HMGB1/HMGB1/β-actin). (a Western blot diagram of RS4:11 and Jurkat cells. (b) Total HMGB1 quantitative data of RS4:11 and Jurkat cells. (c) PARylation-HMGB1 quantitative data of RS4:11 and Jurkat cells. (d) Aceytlation-HMGB1 quantitative data of RS4:11 and Jurkat cells. (B) HMGB1^MT1, HMGB1^MT2 and HMGB1^WT cells were successfully constructed. The lysine residues at amino acids 28, 29, 30, 180, 182, 183, 184 and 185 of HMGB1 in HMGB1^MT1 cells were mutated to alanine, and the glutamate residues at 40, 47 and 179 of HMGB1 in HMGB1^MT2 cells were mutated to alanine. The whole protein of HMGB1^MT1, HMGB1^MT2, HMGB1^WT, HMGB1^NC and Jurkat cells were subjected to immunoprecipitation with anti-HMGB1 antibodies and then subjected to western blotting to detect the expression of lentivirus with anti-Flag antibodies. Tubulin was used as a loading control. (C) Cell lysates were subjected to a pull-down assay with an HMGB1 antibody and immunoblotted with anti-acetylated lysine, anti-poly (ADP-ribosylation) and anti-HMGB1 antibodies. β-actin was used as a loading control. Quantified data are presented (HMGB1/β-actin, PARylation-HMGB1 or Acetylated-HMGB1/HMGB1/β-actin). Data are the mean ± standard deviation of three independent experiments. (a) Western blot diagram of MT1, MT2, WT and NC cells. (b) Total HMGB1 quantitative data of MT1, MT2, WT and NC cells. (c) PARylation-HMGB1 quantitative data of MT1, MT2, WT and NC cells. (d) Aceytlation-HMGB1 quantitative data of MT1, MT2, WT and NC cells. *P<0.05, **P<0.01, Fig. 3Cb-d compared with the untreated group and Fig. 3Ce and f compared with the NC group. HMGB1, high mobility group box protein 1; DNR, daunorubicin; MT, mutant type; WT, wild type; NC, normal control; UT, untreated group.