Figure 8. Analysis of LC3 expression levels. A: western blotting analysis for evaluating the effects of p62 or hsp70 RNA interference on LC3 levels in ARPE-19 cells treated with 5 µM MG-132 and 50 nM bafilomycin. Lane 1. control medium for 36 h; lane 2. MG-132 for 24 h + recover 6 h; lane 3. control medium for 24 h + bafilomycin for 6 h; lane 4. MG-132 + nonsilencing RNA for 24 h + recover 6 h; lane 5. MG-132 + p62 siRNA for 24 h + recover 6 h; lane 6. MG-132 + hsp70 siRNA for 24 h + recover 6 h; lane 7. MG-132 + nonsilencing RNA for 24 h + recover in bafilomycin for 6 h; lane 8. MG-132 + p62 siRNA for 24 h + recover in bafilomycin for 6 h; lane 9. MG-132 + hsp70 siRNA for 24 h + recover in bafilomycin for 6 h; lane 10. control medium for 48 h; lane 11. MG-132 for 24 h + recover 24 h; lane 12. control medium for 24 h + bafilomycin for 24 h; lane 13. MG-132 + nonsilencing RNA for 24 h + recover 24 h; lane 14. MG-132 + p62 siRNA for 24 h + recover 24 h; lane 15. MG-132 + hsp70 siRNA for 24 h + recover 24 h; lane 16. MG-132 + nonsilencing RNA for 24 h + recover in bafilomycin for 24 h; lane 17. MG-132 + p62 siRNA for 24 h + recover in bafilomycin for 24 h; lane 18. MG-132 + hsp70 siRNA for 24 h + recover in bafilomycin for 24 h. α-tubulin staining was used to ensure equal loading of proteins. B: The ratio of a lipidated LC3-II to a nonlipidated LC3-I. Experiments were repeated two independent times.