Fig. 2.

Subcellular localization of NCEH. A: Expression of NCEH and HSL in MPMs was determined by immunocytochemistry using affinity-purified anti-NCEH and HSL rabbit IgG. For detection, cells were incubated with Alexa Fluor® 488-conjugated secondary antibody. As a control, the staining of MPMs with preimmune rabbit IgG as the primary antibody did not produce any fluorescence. Data are representative of three independent experiments. B: The localization of NCEH was analyzed by combined immunocytochemical staining for NCEH (green) and PDI (red) in the presence or absence of 100 µg/ml acLDL. For detection, cells were incubated with either Alexa Fluor 488 or Alexa Fluor 555-conjugated secondary antibodies. Images were acquired with a Bio-Rad MRC 1024 microscope and a 60× objective. Data are representative of three independent experiments. C: S100 and microsomal fractions (10 µg each) of MPMs in the presence or absence of 100 µg/ml acLDL were analyzed by Westen blotting with anti-NCEH antibody. Data are representative of three independent experiments.