Figure 6.

Anthrax LT treatment inhibits c-Jun gene transcription via inactivation of JNKs. A, untreated (M) and LT- and U0126-treated Hep1c1c7 cells were lysed with radioimmune precipitation assay buffer. c-Jun protein in the cell lysates was immunoprecipitated with c-Jun antibody and protein G–conjugated magnetic beads. The levels of phosphorylated and total c-Jun were assessed by Western blotting. Data shown are representative of at least two independent experiments. B, quantitative PCR analysis of c-Jun mRNA expression in Hepa1c1c7 cells treated with LT, U0126, SP600125 (SP), or U0126 + SP600125 for 0, 1, 2, 3, and 4 h. The relative amounts of c-Jun mRNA were normalized by the levels of β-actin mRNA. Shown is one representative experiment, with error bars depicting the standard deviation derived from the results from triplicates for each experimental condition. C, Hepa1c1c7 cells were left untreated or treated with LT, U0126, or U0126+SP600125 for 1, 2, 3, and 4 h. The levels of c-Jun and β-actin were measured by Western blotting. Data shown are representative of at least two independent experiments. The intensities of protein bands were quantified using the Image Studio software of the Odyssey system and normalized by the amounts of β-actin; relative amounts are shown below each lane.