Fig. 4

Pathological α-synuclein mutations (A30P, A53T) alter the α-synuclein-SOD1 interference. a Luciferase activity measurement and western blot analysis (12 μg protein/lane) of H4 cells co-transfected with hGluc tagged wt, A30P or A53T α-synuclein and hGluc tagged SOD1. Data from 3 independent experiments were pooled after normalization to the respective mean of luciferase activity of wt α-synuclein (two tailed, unpaired student’s t-test, n = 9). b Luciferase activity of conditioned medium from cells in a. Shown are the pooled data from 3 independent experiments after normalization to mean of luciferase activity of co-transfected cells with wt α-synuclein (two tailed, unpaired student’s t-test, n = 6-9). c and d Densitometric analysis of western blots of co-transfected H4 cells. SOD1-1, SOD1-2, S1 and S2 were normalized to β-actin (two tailed, unpaired student’s t-test, n = 3). e Representative extracellular complementation assay (ECA) with H4 cell lysates containing 10,98 nM of wt, A30P or A53T α-synuclein or SOD1 is shown. 12 h after combining the lysates, luciferase activity was measured (two tailed, unpaired student’s t-test, n = 3). ECA was repeated 2 times and showed similar results. (n.s. = not significant, * p < 0,05, ** p < 0,005, *** p < 0,0005, **** p < 0,0001) RLU = relative light unit