Figure 6: Transfection of macrophages with the PERK-K618A mutant attenuated E‾/B48 lipoprotein-induced PERK and eIF-2a phosphorylation and cholesterol ester accumulation. Raw 264.7 cells were transfected with a plasmid vector expressing PERK-K618A (K618A) or an empty pEGFP vector. Cells were incubated at 37C for 12 hrs with 20 g/ml of E+/B48 or E‾/B48 lipoproteins or culture medium alone (control). (A-D) Total protein levels of PERK and eIF- 2a and the phosphorylated forms of PERK (PERK-p) and eIF-2a (eIF2a-p) were determined by western blot analysis and expressed relative to -tubulin protein levels. (E and F) Cellular cholesterol contents were determined. Values represent the mean SEM of five separate experiments. *P<0.05 as compared with the control, and P<0.05 as compared with cells transfected with empty pEGFP vector and treated with E‾/B48 lipoproteins.
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