Western blot
After treatment, equal amounts of cells were lysed with RIPA for 60min.
The lysates were quantified by BCA kit (Beyotime, China). The samples
were then separated with SDS-polyacrylamide gels and transferred into
nitrocellulose membranes. After being blocked and cut into proper
sections, membranes were incubated with different primary antibodies
against GAPDH (Abcam, #ab181602), MMP9 (Abcam, #ab76003), MMP13
(Abcam, #ab39012), COX2 (CST, #12282), INOS (Abcam, #ab3523), P53
(Abcam, #ab90363), P21 (Abcam, #ab215971), NF-κB P65 (CST, #8242),
P-NF-κB P65 (Ser536) (CST, #3033), IκB (CST, #4812), and P-IκB (Ser32)
(CST, #2859) at 4 °C overnight. The membranes were then incubated with
luminesced secondary antibodies and detected with the BioRad System.
GAPDH was used as control.