Fig. 7
MMP-9 deficiency enhances LPS-induced IL-6 secretion in RAW264.7 macrophages. RAW264.7 cells were transduced with lentiCRISPRv2 vectors encoding MMP-9-targeting guide RNAs to generate MMP-9 knockout cell lines. Cells were treated with LPS (100 ng/mL) for 24 h. (A) MMP-9 enzymatic activity in the conditioned medium was evaluated by zymography. WT cells treated with PBS served as a negative control; WT cells treated with LPS served as a positive control. (B, C) mRNA levels of Il6 and Tnf were assessed by quantitative PCR from cell lysates, using Rpl19 as an internal control. (D) Secreted IL-6 protein levels in the conditioned medium were quantified by ELISA. Data are expressed as mean ± SEM (n = 3–4 per group). Statistical comparisons were made using unpaired t-tests against LPS-treated WT controls. ns: not significant. m: marker.
