16-18 However, this interplay between Ca and apoptosis has not been studied in the liver in the context of liver regeneration. Therefore, we investigated the role of Ca in the regulation of liver regeneration. Ad, adenovirus; Ad-PV-MITO, parvalbumin–mitochondrial targeting sequence adenovirus; Ad-PV-MITO-GFP, parvalbumin–mitochondrial targeting
Selleck JQ1 sequence–green fluorescent protein adenovirus; AIF, apoptosis-inducing factor; Apaf-1, apoptotic peptidase activating factor 1; ATP, adenosine triphosphate; Bax, B cell lymphoma 2–associated X protein; Bcl-2, B cell lymphoma 2; Bcl-xL, B cell lymphoma extra large; BrdU, bromodeoxyuridine; Camit2+, mitochondrial Ca2+; cDNA, complementary DNA; CT, control; D, day; EGFR, epidermal growth factor receptor; ER, endoplasmic reticulum; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; GFP, green fluorescent protein; IB, immunoblotting; Mcl-1, myeloid cell leukemia 1; MITO-GFP, mitochondrial
targeting sequence–green fluorescent protein; MPO, myeloperoxidase; MTS, mitochondrial targeting sequence; OD, optical density; PCNA, proliferating cell nuclear antigen; PCR, polymerase chain reaction; PH, partial hepatectomy; PI, propidium iodide; PV, parvalbumin; PV-MITO-GFP, parvalbumin–mitochondrial targeting sequence–green fluorescent selleck protein; STA, staurosporine; 99mTc-phytate, phytate labeled with technetium-99m; TNF, tumor necrosis factor. SKHep1 and HEK-293 cell lines were obtained from the American Type Culture Collection (Manassas, VA). Cells were grown at 37°C with 5% carbon dioxide/95% air in Dulbecco’s modified Eagle’s medium supplemented with 1% penicillin-streptomycin and 10% heat-inactivated fetal bovine serum (all learn more from Gibco, Grand Island, NY). The pAc1GFP1-Mito vector, which directs the expression of a GFP-tagged protein to the mitochondrial matrix,
was acquired from Clontech (Mountain View, CA). MitoTracker Red, Rhod-2/AM (fluorescent indicator of mitochondrial Ca2+), the SuperScript first-strand synthesis system for real-time polymerase chain reaction (PCR), PCR SuperMix, Lipofectamine, a caspase-9 detection kit, and antibodies against B cell lymphoma 2–associated X protein (bax), bcl-2, and c-Met were obtained from Invitrogen (Carlsbad, CA). Antibodies against β-actin, anti–γ-tubulin, adenosine triphosphate (ATP), and TNF-α were acquired from Sigma Aldrich (St. Louis, MO). Antibodies against proliferating cell nuclear antigen (PCNA) and epidermal growth factor receptor (EGFR) were obtained from Santa Cruz (Santa Cruz, CA) and Cell Signaling Technology (Boston, MA). Caspase-3 and caspase-8 detection kits were acquired from BD Biosciences (San Jose, CA).