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Fig. 5. PAT proteins suppress ApoB-crescent formation. (A) Overexpression of ADRP and TIP47 suppressed the formation of ApoB-crescents in Huh7 cells. To identify transfected cells, EGFP cDNA was introduced together with empty vector, ADRP cDNA, or TIP47 cDNA. Two days after transfection, cells were treated by 10 µM ALLN for 12 hours and labeled by mouse anti-ApoB (6H12) antibody (red, arrowheads). Asterisks in images indicate the nuclei of EGFP-negative cells. The results from three independent experiments were averaged, and the statistical difference from the control that was transfected with empty vector was examined by the Student's t-test (*P<0.05, **P<0.01). Bars, 10 µm. (B) Knockdown of ADRP and TIP47 by RNAi increased the number of ApoB-crescents. Huh7 cells were examined three days after RNAi without any further treatment. Western blotting of cell lysates (10 µg) confirmed that there was a significant decrease in the levels of the targeted proteins. The results from three independent experiments were averaged, and the statistical difference from the control transfected with control RNA was examined by the Student's t-test (*P<0.001).
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