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. 2014 Sep 25;307(11):G1088–G1099. doi: 10.1152/ajpgi.00210.2014

Fig. 4.

Fig. 4.

A–D: Ex4 mitigates autophagy in steatotic HuH7 cells during HIRI. Lean and steatotic hepatocytes were treated with and without Ex4 during normoxia (HIRI−) and hypoxia (HIRI+). Total cell lysates were subjected to electrophoresis followed by Western blotting with anti-LC3 I/II, p62, high-mobility group protein 1 (HMGB1), beclin-1, autophagy-related protein 7 (ATG7), and β-actin antibodies. A and C: HIRI− and HIRI+. Lanes 1 and 3, lean and FFA with no treatment; lanes 2 and 4, lean and FFA with Ex4 treatment. B and D: relative band density. Experiments were repeated 3 times in triplicate. Values are means ± SD. Ex4 treatment resulted in a statistically significant reduction of autophagy markers in steatotic cells exposed to HIRI. In steatotic cells that were not exposed to HIRI, Ex4 treatment resulted in an increase, rather than a decrease, in autophagy markers. Increase in LC3 II and p62 proteins was statistically significant. While the other proteins did not reach statistical significance, the increasing trend appears clear. *P < 0.05.