Figure 4: TLR2 activation induces bioenergetics changes in RASFC in vitro.

(a) (i) Representative cropped western blot demonstrating expression of PKM2 in RASFC whole cell lysates and nuclear fragments in response to Pam3CSK4 (1 μg/ml) stimulation (n = 3). (ii) Representative photomicrographs demonstrating Pam3CSK4-induced nuclear localisation of PKM2 as indicated by white arrows. Blue arrows represent lack of staining in nuclei under basal conditions. Mag 20X. (n = 3) (b) Representative oxidative phosphorylation (oxygen consumption rate (OCR)) and glycolysis (extracellular acidification rate (ECAR)) Seahorse bioenergetics profiles before and after injections of oligomycin, FCCP and antimycin A for RASFC in the presence and absence of Pam3CSK4 (1 μg/ml). Bar graphs demonstrating (c), baseline respiration (OCR), (d) the ratio of glycolysis: oxidative phosphorylation (ECAR:OCR), (e) ATP synthesis, (f) maximal respiration and, (g) the respiratory reserve in RASFC in response to Pam3CSK4 (1 μg/ml) stimulation (n = 7). Data was analysed using Wilcoxin Signed Rank test and is represented as Mean ± SEM, *p < 0.05, **p < 0.01 significantly different to control.