by Karen Reue, Robert D. Cohen, and Michael C. Schotz

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Evidence for Hormone-Sensitive Lipase mRNA Expression in Human Monocyte/Macrophages by Karen Reue, Robert D. Cohen, and Michael C. Schotz Arterioscler Thromb Vasc Biol Volume 17(12):3428-3432 December 1, 1997 Copyright © American Heart Association, Inc. All rights reserved.

Detection of HSL mRNA in human monocyte/macrophages and THP-1 cells by reverse-transcribed polymerase chain reaction (RT-PCR). a, RNA from human adipose tissue, THP-1 cells (two independent RNA preparations), and monocyte/macrophages (RNA preparations from two independent donor pools) was reverse transcribed and PCR amplified, using β-actin primers or HSL primer set 1 (see Table). Detection of HSL mRNA in human monocyte/macrophages and THP-1 cells by reverse-transcribed polymerase chain reaction (RT-PCR). a, RNA from human adipose tissue, THP-1 cells (two independent RNA preparations), and monocyte/macrophages (RNA preparations from two independent donor pools) was reverse transcribed and PCR amplified, using β-actin primers or HSL primer set 1 (see Table). The 426-bp β-actin product was produced in samples derived from adipose tissue, THP-1 cells, and monocyte/macrophages and visualized by ethidium bromide stain (indicated by upper arrow). The 433-bp HSL product was visualized in adipose tissue, THP-1 cells, and monocyte/macrophages after blotting and hybridization to radiolabeled HSL cDNA (indicated by lower arrow). PCR products were not detected in samples containing water instead of cDNA (right lanes). b, RT-PCR performed on THP-1 cell and monocyte/macrophage RNA samples with primers for scavenger receptor and monocyte chemoattractant protein-1 (MCP). Karen Reue et al. Arterioscler Thromb Vasc Biol. 1997;17:3428-3432 Copyright © American Heart Association, Inc. All rights reserved.

Relative levels of HSL mRNA in human adipose tissue and monocyte/macrophages. Relative levels of HSL mRNA in human adipose tissue and monocyte/macrophages. Semiquantitative reverse-transcribed polymerase chain reaction (RT-PCR) analysis of HSL expression levels in adipose tissue, THP-1 cells, and monocyte/macrophages was performed by monitoring the appearance of amplification product at progressive intervals of the amplification reaction. PCR products were detected by ethidium bromide staining after agarose gel electrophoresis. As shown in the upper portion of the figure, all cDNA samples were diluted 1:30 for amplification with β-actin primers, and PCR products were analyzed after 28, 32, 36, and 40 cycles. In the lower portion, adipose tissue cDNA was diluted 1:40, and THP-1 and monocyte/macrophage cDNA were used without dilution for amplification with HSL primer set 1. Karen Reue et al. Arterioscler Thromb Vasc Biol. 1997;17:3428-3432 Copyright © American Heart Association, Inc. All rights reserved.