Hdac1 antibody

RAW-Blue cells (InvioGen), a mouse macrophage cell line engineered with secreted embryonic alkaline phosphatase (SEAP) reporter system, was used to evaluate LPS-induced NFκB activation in the presence or absence of ILA in vitro. ILA was added to RAW-Blue cells for 1 h before being exposed to LPS. After incubating for 4 h, SEAP levels were quantified by developing supernatant with QUANTI-Blue substrate for 1 h and reading absorption at 620 nm as stated in the manufacturer’s instructions (InvioGen). Caco-2 and HT-29 cells were purchased from ATCC and grown to the specifications. To assess IL-8 production in response to LPS, Caco-2 cells were pre-incubated with ILA for 1 h before overnight exposure to LPS. For gene expression experiments, cells were incubated with ILA in the presence of TNF-α for 1 h or pretreated with AhR or Nrf2 inhibitors for 1 h before 1 h exposure to ILA. Experiments were performed for a total of 6–8 biological replicates. ILA, AhR antagonist CH-223191, Nrf2 antagonist SML1833, TNF-α and LPS 0111:B4 were purchased from Sigma. GAPDH, anti-rabbit HRP and anti-biotin HRP antibodies were purchased from Cell Signaling Technology. AhR antibody was purchased from Invitrogen. HDAC1 antibody was from Santa Cruz Biotechnologies.