Figure 4.
MALT1 inhibition suppresses IL-1β–induced thromboinflammatory gene expression. (A) MALT1 silencing suppresses IL-1β–induced NF-κB activation. HUVECs were transfected with scRNA or MALT1 siRNA (200 nM). After 48 hours, the transfected cells were stimulated with IL-1β for varying times. At the end of treatment, cell lysates were harvested, and the phosphorylation of the P65 subunit of NF-κB was analyzed by immunoblot analysis. The blot was also probed for MALT1 to confirm MALT1 knockdown and GAPDH as a loading control. The left panel shows a representative immunoblot, and the middle and right panels show quantification of MALT1 knockdown and p65 phosphorylation by densitometric analysis of immunoblots. (B) MALT1 silencing suppresses IL-1β–induced expression of TF and VCAM1. HUVECs transfected with scRNA or MALT1 siRNA were treated with a control vehicle (CV) or IL-1β for 6 hours. The cell lysates were probed for VCAM1 and TF expression by immunoblot analysis. The left panel shows a representative immunoblot, and the middle and right panels show quantification of VCAM1 and TF concentrations by densitometric analysis of immunoblots. The concentrations of VCAM1 and TF measured in IL-1β–stimulated cells were taken as 100%, and other values were shown relative to these values. (C-D) (C) MALT1 inhibitor attenuates IL-1β–induced NF-κB activation and (D) expression of VCAM1 and TF. HUVEC were treated with MALT1-specific inhibitor mepazine (20 µM) for 1 hourand then the cells were treated with a control vehcile (CV) or IL-1β for 30 minutes (C) or 6 hours (D). The cell lysates were probed for the phosphorylation of p65 (C) or the expression of VCAM1 and TF (D). The left panel shows a representative immunoblot, and the middle and right panels show densitometric quantification of immunoblot band intensities. The concentrations of p65 were normalized to GAPDH (C). VCAM1 and TF concentrations measured in IL-1β–stimulated cells were taken as 100%, and other values were shown relative to these values. Data are the mean ± standard deviation of 3 independent experiments. ***P < .001.

MALT1 inhibition suppresses IL-1β–induced thromboinflammatory gene expression. (A) MALT1 silencing suppresses IL-1β–induced NF-κB activation. HUVECs were transfected with scRNA or MALT1 siRNA (200 nM). After 48 hours, the transfected cells were stimulated with IL-1β for varying times. At the end of treatment, cell lysates were harvested, and the phosphorylation of the P65 subunit of NF-κB was analyzed by immunoblot analysis. The blot was also probed for MALT1 to confirm MALT1 knockdown and GAPDH as a loading control. The left panel shows a representative immunoblot, and the middle and right panels show quantification of MALT1 knockdown and p65 phosphorylation by densitometric analysis of immunoblots. (B) MALT1 silencing suppresses IL-1β–induced expression of TF and VCAM1. HUVECs transfected with scRNA or MALT1 siRNA were treated with a control vehicle (CV) or IL-1β for 6 hours. The cell lysates were probed for VCAM1 and TF expression by immunoblot analysis. The left panel shows a representative immunoblot, and the middle and right panels show quantification of VCAM1 and TF concentrations by densitometric analysis of immunoblots. The concentrations of VCAM1 and TF measured in IL-1β–stimulated cells were taken as 100%, and other values were shown relative to these values. (C-D) (C) MALT1 inhibitor attenuates IL-1β–induced NF-κB activation and (D) expression of VCAM1 and TF. HUVEC were treated with MALT1-specific inhibitor mepazine (20 µM) for 1 hourand then the cells were treated with a control vehcile (CV) or IL-1β for 30 minutes (C) or 6 hours (D). The cell lysates were probed for the phosphorylation of p65 (C) or the expression of VCAM1 and TF (D). The left panel shows a representative immunoblot, and the middle and right panels show densitometric quantification of immunoblot band intensities. The concentrations of p65 were normalized to GAPDH (C). VCAM1 and TF concentrations measured in IL-1β–stimulated cells were taken as 100%, and other values were shown relative to these values. Data are the mean ± standard deviation of 3 independent experiments. ***P < .001.

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