Figure 2

Induction of MMP-12 mRNA and protein by M.F. in U937 cells and in macrophages. M.F.-infected cells were analyzed by ELISA and real time PCR to detect the M.F. effects on MMP-12. A: 48 h following the infection, the U937 supernatants were collected and analyzed with the ELISA assay. U937 treated with 243 M.F. growth media (v/V) were used as controls for both analysis (ELISA and qRT-PCR). B: RNA samples from U937 cells were collected at the established time points (6 h, 18 h and 24 h) following M.F. infection. MMP-12 expression was analyzed by real time qRT-PCR as described in Materials and Methods. C: Macrophage culture supernatants were harvested 48 h the M.F. infection and tested by ELISA. Uninfected cells and 243 media treated cells were used as controls. D: RNA samples from M.F.-infected macrophages were collected at the following time points: 6 h, 18 h and 24 h. MCP-1 expression was analyzed by real time RT-PCR. Macrophages treated with 243 M.F. growth media (v/V) were used as controls. In both ELISA tests, MCP-1 production was normalized with the Calcein viability assay, as described in the Materials and Methods. Bars denote the standard deviation. The p-values were calculated as unequal variance t-test of Mycoplasma infected cells relative to 243 media control: *p ≤ 0.05; **p ≤ 0.005. The histograms shown are representative of data from three different experiments.