Figure 3.

Defective autophagy and α-synuclein (α-syn) accumulation in T lymphocytes. (a) Western blot analysis of LC3-II and p62 levels in T lymphocytes from patients with SLE cultured with (i) 10% FBS, (ii) 1% FBS and (iii) 1% FBS plus E64d and pepstatin A (Pep A). Blots shown are representative of 10 independent experiments. Densitometry analysis of LC3-II levels (b) and of p62 levels (c) relative to β-actin. Values are expressed as means±S.D. (d) Western blot analysis of α-syn expression in SLE T lymphocytes cultured with (i) 10% FBS, (ii) 1% FBS and (iii) 1% FBS plus E64d and pepstatin A. Blots shown are representative of 10 independent experiments. The bands of α-syn at 14 kDa (monomeric form) and at 70 kDa (aggregate form) are indicated by the arrows. Densitometry analyses of the monomeric form of α-syn at 14 kDa (e) and of the aggregate form of α-syn at 70 kDa (f) are shown. Values are expressed as means±S.D. *P<0.05. (g) Western blot analysis of α-syn in T lymphocytes from healthy donors (HD) and SLE patients cultured with 1% of FBS. T lymphocytes from HD were cultured in the presence or absence of 3-methyladenine (3-MA). Blots shown are representative of three independent experiments. Densitometry analysis of α-syn at 14 kDa (h) and at 70 kDa (i) relative to β-actin is shown. Values are expressed as means±S.D. *P<0.05 versus other experimental conditions. (j) Western blot of α-syn expression in cell lysates obtained from T lymphocytes transfected with control (ctr) siRNA or with ATG5 siRNA. Blots shown are representative of three independent experiments. Densitometry analysis of α-syn at 14 kDa (k) and at 70 kDa (l) relative to β-actin is shown. Values are expressed as means±S.D. *P<0.05 versus other experimental conditions