These authors contributed equally to this work.
Conjugated bile acids activate the sphingosine-1-phosphate receptor 2 in primary rodent hepatocytes†
Article first published online: 30 NOV 2011
Copyright © 2011 American Association for the Study of Liver Diseases
Volume 55, Issue 1, pages 267–276, January 2012
How to Cite
Studer, E., Zhou, X., Zhao, R., Wang, Y., Takabe, K., Nagahashi, M., Pandak, W. M., Dent, P., Spiegel, S., Shi, R., Xu, W., Liu, X., Bohdan, P., Zhang, L., Zhou, H. and Hylemon, P. B. (2012), Conjugated bile acids activate the sphingosine-1-phosphate receptor 2 in primary rodent hepatocytes. Hepatology, 55: 267–276. doi: 10.1002/hep.24681
Potential conflict of interest: Nothing to report.
- Issue published online: 21 DEC 2011
- Article first published online: 30 NOV 2011
- Accepted manuscript online: 19 SEP 2011 09:05AM EST
- Manuscript Accepted: 29 AUG 2011
- Manuscript Received: 26 APR 2011
- National Institutes of Health. Grant Number: R01 DK-057543
Additional Supporting Information may be found in the online version of this article.
|HEP_24681_sm_suppinfofig1.tif||3904K||Online supplemental Fig. 1. Saturation curves of S1P-induced ERK1/2 and AKT activation in primary rat hepatocytes. Primary rat hepatocytes were treated with different amounts of S1P for 20 minutes. The cells were harvested to prepare total cell lysates. The protein levels of phosphorylated ERK1/2 (p-ERK1/2) and AKT (p-AKT) were determined by Western blot analysis and normalized with total ERK1/2 (T-ERK1/2) and total AKT (T-AKT) as described in Materials and Methods. *p<0.05 compared to control group, n=4.|
|HEP_24681_sm_suppinfofig2.tif||6090K||Online supplemental Fig. 2. Expression of S1P1 and S1P2 in hepatocytes. Total RNA was isolated from primary human, rat and mouse hepatocytes. The expression of S1P1 and S1P2 was detected by RT-PCR using specific primers for S1P1 and S1P2. Rat cDNAs of S1P1 and S1P2 were used as positive control (P). The PCR products were purified from DNA agarose gel and further confirmed by DNA sequencing. The representative image of DNA agarose gel was shown.|
|HEP_24681_sm_suppinfofig3.tif||6710K||Online supplemental Fig. 3. Effects of PTX and JTE-013 on activation of ERK1/2 and AKT by S1P in primary rat hepatocytes. (A) Cells were pre-incubated with Pertussis toxin (PTX, 300 ng/ml) for 16 hours or JTE-013 (10 μM) for 30 minutes and then treated with 100 nM S1P for 20 minutes at 37°C. The protein levels of phosphorylated ERK1/2 (p-ERK1/2) and AKT (p-AKT) were determined by Western blot analysis and normalized with total ERK1/2 (T-ERK1/2) and total AKT (T-AKT) as described in Materials and Methods. S1P-induced activation of AKT (A) and ERK1/2 (B) was inhibited by JTE or PTX. *p<0.05 compared to vehicle control; #p<0.05 compared to S1P group, n=4.|
|HEP_24681_sm_suppinfofig4.tif||4934K||Online supplemental Fig.4. Effect of S1P2-shRNA lentivirus on mRNA expression of S1P2 in primary rat hepatocytes. Cells were transduced with control lentivirus or S1P2-shRNA lentivirus for 40 h. Total RNA was isolated. The S1P2 mRNA level was determined by real-time RT-PCR and normalized using ?-actin as internal control. **p<0.01 compared to control lentivirus, n=3.|
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