Figure 6.

Protein O-mannosylation affects the inflammatory response of infected cells. (A) Growth of WT, mutants and complemented strains in hMDM. hMDM were infected at an MOI 1 for 1 h with the indicated strains. Data are presented as the mean +/− SD of four independent experiments performed in duplicate. (B) Production of TNF-α and IL-10 by MHS cells infected by WT, Δpmt and complemented strains. MHS cells were infected at MOI 1:1 for 1 h and TNF-α and IL-10 were evaluated after 5 h or 24 h and 24 or 48 h respectively. Data are representative of two independent experiments performed in triplicate. (C) NK-κB activation in HEK-TLR2 cells incubated with crude extracts from WT and Δpmt M. tuberculosis strains. Activation of NF-κB, evaluated using SEAP activity, following incubation with 500 ng/ml or 50 ng/ml of crude extracts recovered from either bacterial cells or culture medium from the indicated M. tuberculosis strains. PBS 1× or PAM₃CSK₄ (10 ng/ml) were used as negative and positive controls respectively. Data are presented as the mean +/− SD of three independent experiments. (D) NF-κB activation in HEK-TLR2 cells incubated with LprG/LAM complex purified from bacterial cells or culture medium of WT or Δpmt M. tuberculosis strains. HEK-TLR2 cells were incubated with 10 ng/ml or 1 ng/ml of LprG-His and co-purified LAM. SEAP activity was quantified after 24 h. PBS 1× or PAM₃CSK₄ (10 ng/ml) were used as negative and positive controls respectively. Data are presented as the mean +/− SD of at least five independent experiments. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.