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. 2016 Dec 20;6:39553. doi: 10.1038/srep39553

Figure 2. PI16 expression is regulated by ERK5 and modulated by inflammatory cytokines.

Figure 2

To investigate whether ERK5 modulates PI16 expression, ESS was applied to HCAECs for 24 h in the presence of an ERK5 inhibitor BIX02189 (ERK5i; 10 μM). There was a significant reduction in PI16 mRNA (A) and protein (B) expression in HCAECs exposed to laminar shear when compared to the vehicle control (n = 3; *P < 0.05 compared to vehicle control, **P < 0.01 compared to untreated control). We also investigated the regulation of PI16 expression in the presence of inflammatory cytokines, TNFα and IL-1α, which significantly reduced PI16 mRNA (C) and protein (D) expression in HCAECs exposed to ESS for 24 h when compared with the vehicle controls (n = 3; *P < 0.05, **P < 0.01, ***P < 0.001). To exclude the possibility that cytokine treatment affected the adaptation of HCAECs to ESS, a further ‘chronic’ experiment was performed, HCAEC were exposed to ESS for 24 h prior to the first dose of TNFα. 3 doses of TNFα were added to the flow systems at 16 h intervals. TNFα treatment significantly reduced PI16 mRNA (E) and protein (F) expression (n = 3; *P < 0.05, ***P < 0.001) compared to vehicle controls.