Fig. 3

Effects of MT5-MMP deficiency on pro-inflammatory protein levels and IL-1β stability in cortical neural cells. A–C Measurement of MCP-1, IL-1β and IL-6 levels (pg/mL) in primary cultures by ELISA. Note the significant decrease of MCP-1 and IL-1β levels in TgMT5^−/− compared with Tg cells. D–F Measurement of MCP-1, IL-1β and IL-6 levels (pg/mL) in primary cultures by ELISA upon IL-1β treatment (10 ng/mL for 24 h). Black bars represent control (untreated) conditions and grey bars IL-1β treated conditions (10 ng/mL for 24 h). G Immunoblots (top panel) and the corresponding ponceau normalized quantification (lower panels) of IL-1β levels in supernatants and cell lysates after 24 h of incubation with 10 ng/mL IL-1β. Note that the levels of IL-1β were affected in the absence of MT5-MMP. H RT-qPCR analysis of mRNA levels of Mmp2 (upper panel) and Mmp9 (bottom panel) in primary cultures, normalized by Gapdh as housekeeping gene. I Zymogram (upper panel) and the corresponding quantification (bottom panel) of pro-MMP-2 levels in primary neural cultures. J Immunoblot analyses of IL-1β levels after incubation for 24 h at 37 °C in cell conditioned supernatants and lysates with 10 ng/mL IL-1β. Note that none of the the conditioned media modified IL-1β stability after 24 h incubation. Values for A–D, E and H are the mean ± SEM of 6–7 independent cultures by genotype and for F and G 3–6 independent cultures. Values are presented as % of the control. *p < 0.05, **p < 0.01 and ***p < 0.001 between untreated and treated cultures in the same genotype; ^#p < 0.05, ^##p < 0.01 and ^###p < 0.001 between genotypes. ANOVA followed by post hoc Fisher’s LSD test. IB immunoblot, O.D. optical density, A.U. arbitrary units