FIGURE 4.
Elevated p100 levels do not affect CXCL12 expression by HUVEC. A, LZRS- or LZRS-p100-transduced HUVEC were either untreated (C) or incubated with TNF (T; 10 ng/ml), LIGHT (L; 100 ng/ml), or a combination of both cytokines (L+T) for 24 h, and then lysates were immunoblotted using either anti-p100/p52, anti-RelB, or anti-tubulin as a loading control. B, cytosolic (lanes 1–4) and nuclear (lanes 5–8) extracts were made from untransduced HUVEC that were either untreated control (lanes 1 and 5; cont.) or incubated for 24 h with TNF (10 ng/ml) (lanes 2 and 6), and LZRS- (lanes 3 and 7) and LZRS-p100-transduced HUVEC (lanes 4 and 8). Extracts were immunoblotted using anti-p100/p52, anti-tubulin, or anti-histone 3 (H3) as indicated. C, cytosolic (Cyt.) and nuclear (Nuc.) extracts were made from LZRS- and LZRS-p100-transduced HUVEC, and then samples were immunoblotted using either anti-p100/p52, anti-RelB, anti-tubulin, or anti-histone 3 as shown. D, LZRS- (open bar) or LZRS-p100-transduced (filled bar) HUVEC were either untreated (C) or incubated with TNF (T; 10 ng/ml), or LIGHT (L; 100 ng/ml) for 8 h, and then expression of CXCL2 (left) and IL-6 (right) was determined by qRT-PCR. E, LZRS- (open bar) or LZRS-p100-transduced (filled bar) HUVEC were either untreated or incubated with LIGHT (100 ng/ml) for the times indicated, and then CXCL12 expression was determined by qRT-PCR. RQ, relative quantification.