Gapdh antibody

ZFL cells were transfected in six-well plates with miR-145 mimic (final concentration 100 nM), miR-145 hairpin inhibitor (final concentration 100 nM) or miR-control (final concentration 100 nM). After transfection, cells were cultured for 48 hours. Intermediate samples were collected and analyzed by western blot to assess GrnA and MET expression. The lysates were hybridized with the following primary antibodies: the polyclonal anti-GrnA antibody (1:1000), which was produced using the 4MAPS peptide EWEDHKQKKPETQRTTTRPTG (corresponding to residues 244–264 of GrnA) to immunize BALB/c mice (LTK Biolab Incorporation, Taiwan); Met antibody (1:1000, sc-10; Santa Cruz Biotechnologies, USA); and GAPDH antibody (1:1000; AnaSpec, USA).

For immunohistochemistry, 10% neutral buffered formalin-fixed liver tissues were sectioned and hybridized with a PCNA antibody (PC10; Abcam, Cambridge, UK). The liver samples were collected and analyzed by Western blot to assess GrnA, HGF, and c-met expression. The lysates were hybridized with the following primary antibodies: polyclonal anti-GrnA antibody (1:1000), which was produced by immunizing BALB/c mice with 4MAPS peptide EWEDHKQKKPETQRTTTRPTG (corresponding to residues 244–264 of GrnA) (LTK Biolab Incorporation, New Taipei City, Taiwan), HGF antibody (1:1000; Aviva Systems Biology, San Diego, CA, USA), c-met antibody (1:1000, sc-10; Santa Cruz Biotechnologies, Santa Cruz, CA, USA), and GAPDH antibody (1:1000; AnaSpec, Fremont, CA, USA).