Journal: Biomedicines

Article Title: Role of UPR Sensor Activation in Cell Death–Survival Decision of Colon Cancer Cells Stressed by DPE Treatment

doi: 10.3390/biomedicines9091262

Figure Lengend Snippet: BiP is upregulated in the fraction of cells resistant to the DPE treatment. ( A ) RKO and SW480 were treated with DPE (50 or 100 μM) or left untreated as controls. After 48 h, the expression of BiP in living cells or dead cells was evaluated via Western blot analysis. β-Actin was used as the loading control. The histograms represent the means plus S.D. from the densitometric analysis of the ratio between BiP and β-actin. ( B ) The cell viability levels of RKO and SW480 cell lines pre-treated with HA15 (10 μM) and then treated with DPE (50 μM) or left untreated as control (CT) were measured via trypan blue exclusion assay after 48 h of culture. ( C ) RKO and SW480 were pre-treated with HA15 (10 μM) and then treated with DPE (50 μM) or left untreated as control. After 48 h, the expression levels of BiP, CHOP, and Mcl-1 were evaluated via Western blot analysis. β-Actin was used as the loading control. The histograms represent the means plus S.D. from the densitometric analysis of the ratio between the protein and β-actin. Note: ** p < 0.01, * p < 0.05.

Article Snippet: Some experimental cells were plated in 12-well plates, as reported above, then the day after were pre-treated with HA15 (BiP/GRP78 inhibitor) (Sigma-Aldrich, MO, USA, SML2118), 4μ8C (IRE1 RNAse inhibitor) (Sigma-Aldrich, Saint Louis, MO, USA, SML0949), ceapin-A7 (ceapin) (ATF6a signaling blocker) (Sigma-Aldrich, MO, USA, SML2330), or GSK2606414 (GSK) (PERK inhibitor) (Selleckem, Houston, TX, USA, S7307).

Techniques: Expressing, Western Blot, Trypan Blue Exclusion Assay