Figure 3.

KRASmut MIA PaCa-2 cells are more sensitive than KRASwt BxPC-3 to the alteration of the N-glycosylations and the resulting endoplasmic reticulum (ER) stress. (A–D) Protein N-glycosylations were detected in viable MIA PaCa-2 (A,B) and BxPC-3 (C,D) cells after 72 h treatment with 2.5 mM 2-DG −/+ 1 mM mannose (MAN). In particular, tri-/tetra-branched N-glycans were recognized by FITC-conjugated PHA-L. The quantification of the mean fluorescence of independent experiments is displayed in the histograms (A,B), while representative flow cytometric profiles are shown in panels (C,D). (E,F) The expression of the UPR markers was analyzed through Western blot in MIA PaCa-2 (E) and BxPC-3 (F) cells treated with 2.5 mM 2-DG −/+ 1 mM MAN. Actin was used for signal normalization. The images are representative of at least three independent blots. The quantification of the EIF2α phosphorylation and Cleaved Caspase 3, considering all experiments performed, is displayed in the histograms on the bottom. All data represent the mean ± SEM of at least three independent experiments. * p < 0.05, ** p < 0.01 and *** p < 0.001 (One-way ANOVA), untreated vs. treated where not specifically indicated.