spacer gif spacer gif spacer gif spacer gif spacer gif
QUICK SEARCH:   [advanced]


spacer gif
     Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents    

Right arrow Help viewing high resolution images
Right arrow Return to article
(Downloading may take up to 30 seconds.
If the slide opens in your browser, select File -> Save As to save it.)
Click on image to view larger version.


Figure 1


Fig. 1. ApoB lipidated by MTP activity is necessary for ApoB-crescent formation. (A) Incubation with MTP inhibitors (100 nM BAY13-9952 or WM1159) for 12 hours significantly decreased the number of ApoB-crescents in Huh7 cells. Triple labeling for ApoB (red), LD (green), and nucleus (blue) is shown. ApoB was labeled by a mouse anti-ApoB (6H12) antibody. The same result was obtained in HepG2 cells (data not shown). The number of ApoB-crescent-positive cells was reduced by the MTP inhibitors (left half of the bar graph), and it did not increase even after further treatment with 10 µM ALLN for 12 hours (right half of the bar graph). More than ten pictures were randomly taken, and the ratio of ApoB-crescent-positive cells was quantified. The results from three independent experiments were averaged, and the statistical difference from the control was examined by the Student's t-test (*P<0.001). Bars, 10 µm. (B) Knockdown of MTP or ApoB by RNAi reduced the number of ApoB-crescents in Huh7 cells. The ratio of ApoB-crescent-positive cells was measured three days after RNAi, and after treatment with 10 µM ALLN for the final 12 hours. For western blotting, an equal amount (30 µg) of each cell lysate was electrophoresed. The results from three independent experiments were averaged, and the statistical difference from the control was examined using the Student's t-test (*P<0.001). (C) The ApoB-crescent was increased by ALLN treatment even after protein translation was inhibited using 5 µg/ml cycloheximide (CHX). The results from three independent experiments were averaged, and the statistical difference was examined by the Student's t-test (*P<0.01). (D) Treatment with either 0.4 mM DHA complexed to BSA or 1 µg/ml cyclosporin A for 12 hours considerably increased ApoB-crescent formation. Representative immunofluorescence micrographs using a mouse anti-ApoB (6H12) antibody are presented. The ratio of ApoB-crescent-positive cells is shown in the bar graph. The results from three independent experiments were averaged, and the statistical difference from the controls was examined by a Student's t-test (*P<0.01, **P<0.001).





Right arrow Return to article