Fig. 4.

Peroxisome proliferator-activated receptor (PPAR) γ activation played a pivotal role in killing bladder cancer cells. (A) Cells were treated for 48 h with 25–400 μM rosiglitazone and cell viability was measured using CCK-8 assay. Mean ± SD, n = 6. (B) Western blotting for PPARγ, glucose-regulated protein (GRP) 78, endoplasmic reticulum resident protein (ERp) 44, and acetylated histone. Cells were treated for 48 h with 50–200 μM rosiglitazone. Actin was used for the loading control. Representative blots are shown. (C) Cells were treated for 48 h with 50–100 μM rosiglitazone and/or 5–20 nM romidepsin and cell viability was measured using CCK-8 assay. Bars represent mean ± SD, n = 6. (D) Cells were treated for 48 h with 100 μM rosiglitazone and/or 20 nM romidepsin and reactive oxygen species production was measured by dihydroethidium (DHE) staining using flow cytometry. 10,000 cells were counted. Bar graphs show the relative DHE fluorescence intensity. Data are expressed as mean ± SD from three independent experiments. *p = 0.0495; N. S., not significant. (E) Cells were treated for 48 h with 100 μM rosiglitazone and/or 20 nM romidepsin. Apoptotic cells were detected by annexin-V assay using flow cytometry. 10,000 cells were counted. Bar graphs show the percentages of apoptotic cells. Data are expressed as mean ± SD from three independent experiments. FITC, fluorescein isothiocyanate; 7-AAD, 7-amino-actinomycin D. *p = 0.0495. (F) Western blotting for PPARγ, GRP78, ERp44, and acetylated histone. Cells were treated for 48 h with 100 μM rosiglitazone and/or 20 nM romidepsin. Actin was used for the loading control. Representative blots are shown.