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Additional Supporting Information may be found in the online version of this article.

FilenameFormatSizeDescription
hep26625-sup-0001-suppfig1A.tif1521KSupporting Figure 1. Effect of PKC inhibitor treatment on covalent binding and PKC activity. A) Effect PKC inhibitors on covalent binding. Primary hepatocytes were treated with APAP 25mM. After 2 h APAP was removed and cells were treated with different PKC inhibitors [Ro 31-8425 (5μM), Go6983 (10μM) or Go6976 10 μM]. After 2 hours of PKC inhibitor treatment, hepatocytes were washed and scraped with lysis buffer. Western blot analyses were performed with antisera against NAPQI protein adducts in whole cell lysate to assess covalent binding to liver proteins. B) Go6983 and Go6976 treatment (post-treatment) inhibits PKC activity. PKC activity was assessed in hepatocyte lysate by western blotting using an antibody that recognizes proteins phosphorylated by PKC.
hep26625-sup-0002-suppfig1B.tif1521KSupporting Figure 1. Effect of PKC inhibitor treatment on covalent binding and PKC activity. A) Effect PKC inhibitors on covalent binding. Primary hepatocytes were treated with APAP 25mM. After 2 h APAP was removed and cells were treated with different PKC inhibitors [Ro 31-8425 (5μM), Go6983 (10μM) or Go6976 10 μM]. After 2 hours of PKC inhibitor treatment, hepatocytes were washed and scraped with lysis buffer. Western blot analyses were performed with antisera against NAPQI protein adducts in whole cell lysate to assess covalent binding to liver proteins. B) Go6983 and Go6976 treatment (post-treatment) inhibits PKC activity. PKC activity was assessed in hepatocyte lysate by western blotting using an antibody that recognizes proteins phosphorylated by PKC.
hep26625-sup-0003-suppfig2.tif1521KSupporting Figure 2. Effects of silencing PKC-α and PKC-ε in the liver. A) Silencing PKC-α using ASO increase CYP2E1 levels in the liver. Mice were treated with PKC-α or control ASO for 2 weeks. B) PKC-ε knockout mice are not protected from APAP-induced liver injury. Mice were treated with acetaminophen (300 mg/kg; ip). Mice were sacrificed at 24 hours, and ALT levels were measured. N = 5 mice per group.

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