Scramble rna

Differentiated THP-1 cells were transfected with 30 nM small interfering RNA (siRNA) for LXRα (Santa Cruz, Dallas, TX, USA, #sc- 38828), LXRβ (Santa Cruz, #sc-45316), SREBP1 (Santa Cruz, Dallas, TX, USA, #sc-36557), SREBP2 (Santa Cruz, Dallas, TX, USA, #sc-36559) or Scramble RNA (Santa Cruz, Dallas, TX, USA, #sc-37007) using the Lipofectamine RNAiMAX Transfection Reagent according to the manufacturer’s instructions (Thermofisher, Carlsbad, CA, USA, #13778075). The knockdown efficiency was confirmed by qPCR, 24 h post-transfection (Figure S5A–C). The transfected cells either remained untreated or were treated with 20 µg/mL oxLDL for 24 has described earlier. The cells were rested for 4 days and restimulated with 10 µg/mL of Pam3cys. Additionally, the concentrations of IL-6 and TNFα in the supernatant were measured using ELISA.

STAT1 siRNA and scramble RNA were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Transfection of siRNA was performed using Lipofectamine RNAimax (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instructions. Plasmids including eGFP-STAT1, eGFP-STAT1Y701F, eGFP-STAT1S727A, Flag-STAT1β were purchased from Addgene (Cambridge, MA, USA). Flag-tagged, constitutive-active STAT1 (or STAT1C) cloned into the backbone of pcDNA3.1 was a gift from Dr. Toru Ouchi (Roswell Park Cancer Institute, University at Buffalo, NY, USA). The constitutively active MEK-1 (HA-ca-MEK) vector was a gift from Dr. Nathalie Rivard (Université de Sherbrooke, Québec, CA). The proteasome inhibitor N-carbobenzoxyl-L-leucinyl-L-norleucinal (MG132) was purchased from Calbiochem (La Jolla, CA, USA) and 5-Aza-2′-deoxycytidine (5-Aza) was purchased from Sigma (St Louis, MO, USA). U0126, a Mitogen-activated protein kinase 1 (MEK1) inhibitor was purchased from Sigma (St Louis, MO, USA).