Journal: The Journal of Physiology

Article Title: Bile acids induce necrosis in pancreatic stellate cells dependent on calcium entry and sodium‐driven bile uptake

doi: 10.1113/JP272774

Figure Lengend Snippet: A , mRNA expression of selected genes encoding bile acid transporters in human pancreatic stellate cells (hPSCs) and human hepatocytes. PCR products for the sodium–taurocholate cotransporting polypeptide (NTCP, slc10A1) and sodium independent transporters, OATP4A1 ( slco4A1 ) and OATP1B3 (slco1B3) , are present in human hepatocytes; hPSCs express slc10A1 and slco4A1 , but not slco1B3 . B , immunoblotting shows expression of NTCP in different tissues; lines from the left: mass marker, hPSC, four positive controls (human liver and mouse pancreas, liver and kidney), two negative controls (mouse spleen and lung). C , immunohistofluorescence (IHF) staining for NTCP in fixed mouse tissue sections: the pancreas (upper panel) and the liver (lower panel). From the left: DAPI (blue), NTCP (white), overlay images (white arrows indicate PSCs), and transmitted light images. Insets show corresponding staining in controls without primary antibody. D , double IHF staining for NTCP and the bradykinin receptor B2 (BDKRB2). From the upper left: DAPI (blue), NTCP (green), BDKRB2 (red), fluorescence overlay (PSCs are indicated by white arrows). E and F , the comparison of homological sequences of three Na + ‐dependent mouse transporters (NTCP, ASBT and SOAT) with immunogens used for generation of the antibodies for immunoblotting ( E ) and IHF ( F ). Red letters represent amino acids identical to those in the immunogen sequence; blue letters are similar amino acids, and black letters show amino acids that are different. Only mouse NTCP sequences share high sequence identity with the immunogens.

Article Snippet: Human liver tissue lysate was obtained from Novus Biologicals, Littleton, CO, USA.

Techniques: Expressing, Western Blot, Marker, Immunohistofluorescence, Staining, Fluorescence, Comparison, Sequencing