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. 2002 Sep;70(9):4841–4850. doi: 10.1128/IAI.70.9.4841-4850.2002

FIG. 3.

FIG. 3.

Effect of PDTC (50 μM) or TPCK (100 μM) on O. tsutsugamushi-induced MCP-1 mRNA level and NF-κB in HUVEC. (A) MCP-1 mRNA level was analyzed using an RNase protection assay with total RNA samples that were prepared from uninfected cells (Mock), cells infected with O. tsutsugamushi for 3 h (OT), and infected cells in the presence of PDTC (OT + PDTC) or TPCK (OT + TPCK). (B) EMSA was performed to analyze the activation of NF-κB using nuclear extracts prepared from HUVEC treated for 2 h with medium (Mock), L-929 cell lysate (Lysate), or O. tsutsugamushi. Nuclear extracts from cells pretreated with PDTC (OT + PDTC) or TPCK (OT + TPCK) for 1 h before infection with O. tsutsugamushi were also analyzed. (C) In the supershift assay, nuclear extracts from HUVEC were preincubated with antibodies against the p65 subunit of NF-κB.