Figure 2. NLRP12 is important for NF-κB activation, cytokine/chemokine production, HDAC expression and MAPK activation in human macrophages following K. pneumoniae infection.

A-B. Human monocyte-derived macrophages (0.5 ×10⁶) were transfected with 40 nM NLRP12 siRNA or scrambled siRNA (nonspecific; NS) for 48 h. Cells were then infected with 1 MOI of K. pneumoniae for 3 or 6 h and NF-κB activation was measured by western blotting (A), and cytokine/chemokine levels in supernatants (B) were measured by sandwich ELISA. Representative blots are shown from three independent experiments with identical results. Densitometry was performed using Gel Digitizing Software and normalized to GAPDH. C-D. Human macrophages were transfected with siRNA and then infected with K. pneumoniae in a similar manner, and cell lysates were used to determine HDAC expression and MAPK activation by western blotting. Representative blots are shown from three independent experiments with identical results. Densitometry was performed using Gel Digitizing Software and normalized to GAPDH. Means ± SE values were obtained from three separate experiments. NS, non-specific. *, p<0.05; **, p<0.01;***, p<0.001. Error bars represent SE.