Figure 5
miR-27a counteracts mitoxantrone-induced ICD through the same UPR pathway. (a) Immunoblot analysis of PERK, p-PERK, eIF2α, p-eIF2α and GRP78 in time-course experiments of HCT116 CRTL, miR27a_KD and miR27a_OE cells treated with 1 μM MTX; phosphorylation was referred to equivalent amounts of the non-phosphorylated forms of the proteins in all cells and then to β-actin used as a loading control. Quantification of the bands in basal and in the kinetic conditions is reported. (b) CRT cell surface expression in HCT116 CRTL, miR27a_KD and miR27a_OE cells untreated or treated for 12 h with 20 mM LY-294002, alone or in combination with 1 μM MTX for the last 6 h. Densitometric analysis was carried out after normalization to E-cadherin. *P⩽0.05; ** P⩽0.01 (two-tailed Student’s t-test) versus HCT116 CRTL cells; #P⩽0.05; ##P⩽0.01 (two-tailed Student’s t-test) versus MTX-untreated cells. The values are representative of five experiments
