SR141716 down-regulates <i>pFLJ</i> expression and inhibits fat deposition in cultured intramuscular adipocytes.

<p>(A and B) qPCR analysis of pFLJ expression (A) and measurement of total triglyceride (B) in cultured adipocytes 24 and 48 hours after SR141716 treatment. The qPCR values are shown as expression fold changes after normalization against the control 18s rRNA. Data are presented as means ± standard error. Gene ID was as shown. Cells were stained with Oil-Red O to determine lipid accumulation (total triglyceride). *: P<0.05, **: P<0.01.</p

List of genes encoding transcription factors and signaling molecules differentially expressed in <i>longissimus dorsi</i> muscle in Jinhua and Landrace pigs at d90.

<p>List of genes encoding transcription factors and signaling molecules differentially expressed in <i>longissimus dorsi</i> muscle in Jinhua and Landrace pigs at d90.</p

pFLJ functions as a positive regulator of fat deposition in intramuscular adipocytes.

<p>(A) Cell images to verify transfection efficiency. Cells were transfected with pSilencer TM 4.1-CMV neo plasmids carrying the sequences fs1, fs2 and fs3. Transfection efficiency was assessed by expression of the reporter gene <i>EGFP</i> (green color) harbored by the plasmid. (B) qPCR analysis of <i>pFLJ</i> expression in cultured adipocytes 24 hours after siRNA treatment. fs1, fs2 and fs3: pFLJ specific siRNAs; ns: negative control siRNA. (C) qPCR analysis of <i>FAS</i>, <i>ACC</i>, <i>ATGL</i> and <i>HSL</i> in cultured adipocytes treated with fs1 siRNA. The qPCR values are shown as expression fold changes after normalization against the control 18s rRNA. Data are presented as means ± standard error. *: P<0.05, **: P<0.01 (C and D) Measurement of total triglyceride (as before) in the cultured adipocytes or free glycerol (the free glycerol release was normalized to total cellular protein and expressed relative to the control group) in the culture medium 36 hours after treatment with fs1 siRNA. ^ab means every two columns with different letters are significantly different (P<0.05).</p

List of representative genes for adipogenesis and myogenesis differentially expressed in <i>longissimus dorsi</i> muscle in Jinhua and Landrace (L) pigs at d90.

<p>List of representative genes for adipogenesis and myogenesis differentially expressed in <i>longissimus dorsi</i> muscle in Jinhua and Landrace (L) pigs at d90.</p

List of representative genes for adipogenesis and myogenesis differentially expressed in <i>longissimus dorsi</i> muscle in Jinhua and Landrace (L) pigs at d30.

<p>List of representative genes for adipogenesis and myogenesis differentially expressed in <i>longissimus dorsi</i> muscle in Jinhua and Landrace (L) pigs at d30.</p

Summary of the number of genes up- or down-regulated in <i>longissimus dorsi</i> muscles in Jinhua or Landrace pigs at age of d90 and d150.¹

1<p>Number of genes was obtained by comparing the expression profiles between d30 and d90 or d30 and d150 in each breed. Details are listed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0053181#pone.0053181.s002" target="_blank">Tables S2</a> (d90-up in Jinhua pigs), S3 (d90-down in Jinhua pigs), S4 (d150-up in Jinhua pigs), S5 (d150-down in Jinhua pigs), S6 (d90-up in Landrace pigs), S6 (d90-down in Landrace pigs), S7 (d150-up in Landrace pigs), and S8 (d150-down in Landrace pigs).</p

The Jinhua breed has a higher IMF content than the Landrace breed.

<p>(A) Oil Red O staining of <i>longissimus dorsi</i> muscles in Jinhua and Landrace pigs, respectively. Oil Red O stained IMF displayed a red color. (B) Comparison of IMF contents in <i>longissimus dorsi</i> muscles in Jinhua and Landrace pigs at the age of d30, d60, d90, d120 and d150, respectively. Pigs were slaughtered at around the age of d30, d90 and d150 (nine individuals per stage) and d60 and d120 (three individuals per stage) for each breed. Data are presented as means ± standard error. **P<0.01. Scale bars, 100 µm.</p