Fig. 4. Intracellular signaling pathways in Neisseriae internalization.

BEAS-2B cells incubated in antibiotic-free and FBS-free culture medium with A) SB03580 (black bars), B) U0126 (gray bars), C) SP600125 (white bars) and D) parthenolide (PA) (striped bars) (25 μg/ml) for 1h followed by addition of NM-Nlac PorB] (striped bars), NM (gray bars) and NL (white bars), MOI 100, 2h. Intracellular bacteria are expressed as percent relative to control cells in the absence of inhibitors (medium/DMSO as inhibitors diluent control) (dashed line) ± SEM. SB03580: **p = 0.0084 by one sample t test; U0126: *p = 0.045 and ***p = 0.0002 as above, and *p = 0.026 by Mann Whitney test; SP600125: *p = 0.012, **p = 0.0011 and ****p < 0.0001 as above; PA: **p = 0.0013 and ****p < 0.0001 as above. E) BEAS-2B cells incubated with inhibitors as above prior to stimulation with PorB^WT (10 μg/ml, 24h) or Pam₃CSK₄ (200 ng/ml, 24h) and addition of NM-[Nlac PorB] (striped bars), NM (gray bars) and NL (white bars) MOI 100, 2h. Intracellular bacteria are expressed as percent relative to cells stimulated in the absence of inhibitors (dashed line) ± SEM. SB03580: ***p = 0.0003 and 0.0005 (PorB^WT); **p = 0.0042 and ****p < 0.0001 (Pam₃CSK₄) by one sample t test. U0126: ***p = 0.0005 and 0.0007, and ****p< 0.0001 (PorB^WT); **p = 0.0027, ***p = 0.0004 and ****p < 0.0001 (Pam₃CSK₄) as above. SP600125: **p = 0.0011, ***p = 0.0004 and ****p < 0.0001 (PorB^WT); *p = 0.015 and ****p < 0.0001 (Pam₃CSK₄) as above. PA: *p = 0.012 and ****p < 0.0001 (PorB^WT); ***p = 0.0001 and ****p < 0.0001 (Pam₃CSK₄) as above.