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Figure 1. Analysis of four different hPRLR mRNA isoforms (L, I, S1<sub>a</sub>, and S1<sub>b</sub>) in human breast adipose tissue (Breast AT), sc abdominal adipose tissue (WAT), and ovary using RT-PCR, followed by Southern blot analysis. A, Detection of L- (856 bp) and I-PRLR (284 bp) by RT-PCR using the hPRLRa- and hPRLRb-primer, followed by Southern blot analysis using a <sup>32</sup>P-labeled probe (nucleotides 1148–1391) that recognizes both L- and I-PRLR. B, Detection of S1<sub>a</sub>- (460 bp) and S1<sub>b</sub>-PRLR (306 bp) by RT-PCR using the hP23- and hP44-primer (23 ), followed by Southern blot analysis using a <sup>32</sup>P-labeled probe (nucleotides 1049–1391) that recognizes both S1<sub>a</sub>- and S1<sub>b</sub>-PRLR. From: Identification of Functional Prolactin (PRL) Receptor Gene Expression: PRL Inhibits Lipoprotein Lipase Activity in Human White Adipose Tissue J Clin Endocrinol Metab. 2003;88(4): doi: /jc J Clin Endocrinol Metab | Copyright © 2003 by The Endocrine Society
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Figure 2. Immunoblot analysis of hPRLR protein expression in human breast adipose tissue (Breast AT), sc abdominal adipose tissue (WAT), and the ovary. A, A hPRLR antibody that recognizes an epitope in the extracellular domain of the hPRLR was used (7770–6010; Biogenesis). Positions of the M<sub>r</sub> 90,000 band (L-PRLR), M<sub>r</sub> 50,000 band (I-PRLR), approximately M<sub>r</sub> 40,000 band, and approximately M<sub>r</sub> 35,000 band (possibly the S1<sub>a</sub>- and S1<sub>b</sub>-PRLR) are indicated. B, An antibody that recognizes an epitope specific to the human I-PRLR was used (34–48000; Zymed Laboratories, Inc. Corp.). Position of the M<sub>r</sub> 50,000 band (I-PRLR) is indicated. From: Identification of Functional Prolactin (PRL) Receptor Gene Expression: PRL Inhibits Lipoprotein Lipase Activity in Human White Adipose Tissue J Clin Endocrinol Metab. 2003;88(4): doi: /jc J Clin Endocrinol Metab | Copyright © 2003 by The Endocrine Society
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Figure 3. The direct effects of PRL on LPL activity in human adipose tissue cultured in vitro. Human adipose tissue was incubated for 6 d. A, During the last 24 h, the following hormonal treatments were studied: control (no hormone added), hPRL (500 ng/ml), and hGH (100 ng/ml). B, During the last 24 h, the following hormonal treatments were studied: control (no hormone added), cortisol (1 μm), cortisol plus hPRL (1 μm cortisol, and 500 ng/ml hPRL), and cortisol plus hGH (1 μm cortisol and 100 ng/ml GH). For each incubation experiment, adipose tissue from one subject was used, and in each hormonal treatment, there were duplicate samples. The LPL activity was analyzed as previously described (24, 25 ). The average LPL activity in the control was set at 100%. Results are expressed as the mean ± sem (each bar represents eight individual cultures derived from four patients), and bars with different superscripts are significantly different from each other (P < 0.05), using one-way ANOVA, followed by Student-Newman-Keuls multiple range test. From: Identification of Functional Prolactin (PRL) Receptor Gene Expression: PRL Inhibits Lipoprotein Lipase Activity in Human White Adipose Tissue J Clin Endocrinol Metab. 2003;88(4): doi: /jc J Clin Endocrinol Metab | Copyright © 2003 by The Endocrine Society
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