Sc 390967

The cells were lysed in lysis buffer (Beyotime Institute of Biotechnology, Shanghai, China) and centrifuged at 10,000 x g for 10 min at 4°C, and placed on ice for 30 min to obtain the supernatant. The protein concentration was determined using a BCA kit (Beyotime Institute of Biotechnology). After being separated by 10% SDS-PAGE (40 µg protein was used), the proteins were transferred onto nitrocellulose membranes (EMD Millipore, Bedford, MA, USA) and blocked for 1 h. The membranes were incubated overnight at 4°C with anti-EGR3 (1:500, sc-390967, Santa Cruz Biotechnology, Inc., Dallas, TX, USA), anti-IL-6 (1:1,000, ab9324, Abcam, Cambridge, USA), anti-IL-8 (1:1,000, ab18672, Abcam), anti-IL-1β (1:1,000, ab156791, Abcam) and anti-β-actin (1:1,000, sc-517582, Santa Cruz Biotechnology, Inc.) and incubated with secondary antibodies (1:1,000, HRP-labeled goat anti-mouse IgG, A0216, Beyotime Institute of Biotechnology) for 2 h at room temperature. Proteins were visualized with an ECL kit (GE Healthcare, Chicago, IL, USA). The Gray value was detected by ImageJ 1.48 software.