Effect of BXL-01-0029 onto TNFα-treated Hfsmc.

A, B, Western blot analysis was performed to assess JNK (A) and NF-kB (B) activation after 12 min stimulation with TNFα (10 ng/ml), with or without BXL-01-0029 (10−8 M). TNFα induced JNK and NF-kB phosphorylation; BXL-01-0029 impaired JNK activation and prevented NF-kB phosphorylation (upper A and B); total JNK and NF-kB were used as loading control (middle A and B). Lower A and B report densitometric analysis (*P<0.05 vs. control, #P<0.05 vs. TNFα-treated cells) and results are expressed as ratio phosphorylated/total protein arbitrary units vs. TNFα-treated cells, taken as 1 (mean±S.E.). C, Bio-Plex protein analysis assessed ERK1/2 activation after 12 min stimulation with TNFα (10 ng/ml), with or without BXL-01-0029 (10−8 M). TNFα and BXL-01-0029 induced ERK1/2 phosphorylation (*P<0.05 vs. control); their combination enhanced this effect (#P<0.05 vs. TNFα-treated cells). Results are expressed as fold of relative p- to tot-ERK1/2 expression vs. TNFα-treated cells, taken as 1 (mean±S.E.). Data were obtained from two experiments with different cell preparations. D, TNFα-induced CXCL10 secretion, taken as 100%, was significantly reduced by BXL-01-0029 (10−8 M) after 24 h incubation; *P<0.05 vs. TNFα-induced secretion. Results (mean±S.E.) are derived from five experiments, using distinct cell preparations.