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. Author manuscript; available in PMC: 2021 Sep 4.
Published in final edited form as: Int Immunopharmacol. 2020 May 22;85:106612. doi: 10.1016/j.intimp.2020.106612

Figure 6. Imperatorin suppressed iNOS by inhibiting ERK-MAPK pathway.

Figure 6.

(A) Human OA chondrocytes were treated with IL-1β in the presence or absence of Imperatorin and the activation of NF-κB, p-38, JNK, MEK1/2, ERK1/2, c-Fos and c-Jun was investigated by Immunoblotting. Immunoblot (B and C) Band intensities of MEK1/2 and ERK1/2 were quantified by ImageJ. (D) Primary human OA chondrocytes were pre-treated with PD98059 (5μM) to inhibit MEK/ERK followed by treatment with IL-1β (1 ng/ml). Culture supernatants were harvested to determine NO level and (E) Chondrocytes were harvested for RNA isolation followed by qPCR analysis for iNOS expression. β-actin was used as endogenous normalization control. (F) Translocation of c-Fos to nucleus was analyzed by immunofluorescence using c-Fos-specific antibody. (G) Translocation of c-Jun to nucleus was analyzed by immunofluorescence using c-Jun-specific antibody. Nuclei were counterstained with DAPI and images were captured by Olympus FV1000 confocal microscope (60X magnification).