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Figure 2


Fig. 2. SiRNA to PLD1 reduces the formation of cytosolic lipid droplets in NIH 3T3 cells. (A) Immunoblots of PLD1 in untransfected cells and of PLD1, GRP78 or ß-actin in cells transfected with PLD1 siRNA or a control siRNA. (B,C) Effect of PLD1 siRNA or a control siRNA on lipid droplet formation in cells treated with oleic acid for 7 hours. Two days after the transfection with the siRNA, the cells were stained with Oil Red O. (B) Micrographs showing staining. Bar, 10 µm. (C) BioPix software was used to calculate the total area of Oil Red O-stained droplets per cell; values are mean ± s.d. of all cells in ten randomly selected micrographs from each group (P<0.001, Mann-Whitney rank sum test). (D) Effect of PLD1 siRNA (triangles) and control siRNA (squares) on the accumulation of radioactive triglycerides in oleic acid-treated NIH 3T3 cells after incubation with [3H]palmitic acid for the indicated times. Values are mean ± s.d. from cells from three culture dishes. (E) Influence of PLD1 siRNA on the amount of ADRP in the cell. Cells were treated with siRNA as in A and the cell lysate was blotted against an antibody to ADRP.





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