Figure 4.

Etoposide induces mitochondrial unfolded protein response (UPR^mt). HCT116 cells were treated with etoposide (Etop; 10 μM) for various time periods. (a and b) Whole cell lysates were prepared and subjected to western blotting for mentioned mitochondrial UPR proteins. Actin serves as loading control. (c) HCT116 cells treated with etoposide (Etop; 10 μM) for 24 h. Equal amounts of total RNA were subjected to RNA-Seq library preparation followed by sequence reads (80 millions). Expression levels of CHOP, CEBP, ClpP, DnaJ, Hsp10, Hsp60, Hsp70, Hsp90, and HSF-2 are presented. (d) HCT116 cells treated with etoposide (Etop; 10 μM) for 24 h. Total RNAs were isolated from treated and untreated cells and equal amounts of RNA was used for real-time PCR analysis for mentioned UPR genes including the expression levels of CHOP, CEBP, ClpP, DnaJ, Hsp10, Hsp60, Hsp70, Hsp90, and HSF-2. Data are mean±S.D., n=3