Skip to main content
. Author manuscript; available in PMC: 2020 May 1.
Published in final edited form as: Eur Spine J. 2019 Mar 7;28(5):993–1004. doi: 10.1007/s00586-019-05910-9

Fig. 4. Reduced serum concentration executes disc cellular autophagy.

Fig. 4

(A) Western blotting for light chain 3 (LC3), high mobility group box 1 (HMGB1), and p62/sequestosome 1 (p62/SQSTM1) in rabbit and human total protein extracts after 48-h culture in Dulbecco’s modified Eagle’s medium (DMEM) with 0% and 10% fetal bovine serum (FBS). Actin was used as a loading control. Immunoblots shown are representative of four experiments with similar results (n = 4). (B) Changes in protein expression of LC3-II, HMGB1, and P62/SQSTM1 relative to actin in species analysis. Data are mean ± SD (n = 4). Two-way analysis of variance (ANOVA) with the Tukey-Kramer post-hoc test was used. *P < 0.05. **P < 0.01. (C) Time-course Western blotting for LC3, HMGB1, p62/SQSTM1, and actin in total protein extracts after 0, 12-, 24-, and 48-h culture in DMEM with 0%, 1%, or 10% FBS. Immunoblots shown are representative of four experiments with similar results (n = 4). (D) Changes in protein expression of LC3-II, HMGB1, and p62/SQSTM1 relative to actin in time-course analysis. Data are mean ± SD (n = 4). Two-way ANOVA with the Tukey-Kramer post-hoc test was used. *P < 0.05. **P < 0.01. (E) Western blotting for LC3 and actin to assess LC3 turnover using 15-μM chloroquine in total protein extracts after 48-h culture in DMEM with 0%, 1%, or 10% FBS. Immunoblots shown are representative of four experiments with similar results (n = 4). (F) Changes in protein expression of LC3-II relative to actin in LC3 turnover assay. Data are mean ± SD (n = 4). Two-way ANOVA with the Tukey-Kramer post-hoc test was used. *P < 0.05. **P < 0.01.