Fig. 5

Evaluation of autophagy, apoptosis, and citrullination after in vitro treatment with etanercept in PBMCs from patients with RA. a LC3-II levels in PBMCs purified from patients with RA after in vitro treatment with etanercept at concentration of 15 μg/ml. Where indicated, cells were starved (1% FBS) or incubated with TNFα for 4 h, and then etanercept was added to the culture for 24 h. Densitometry analysis obtained in five independent experiments is also shown, **P < 0.01, *P < 0.05. b Flow cytometry analysis of apoptosis after in vitro treatment with etanercept as previously described. Results, referred to five independent experiments in PBMCs isolated from patients with RA, are expressed as percentage of AV-positive apoptotic cells. Representative dot plots are shown (PI on y-axis vs. AV on x-axis), **P < 0.01. c Levels of citrullinated proteins in PBMCs isolated from patients with RA treated with etanercept in vitro. Densitometric analysis is also shown. Data are obtained from five independent