Figure 4.

Assessment of the OXPHOS capacity in primary nerve cultures genetically deleted for PSs. Oxygen consumption rate (OCR) was evaluated by using the Seahorse XF96 bioenergetic analyzer. Experiments were carried out in primary DIV11 neuronal cultures (left panel) and in primary DIV24 astrocyte cultures (right panel). Cell conditions were wild-type non-infected (Ct) vs. PS2KO; control infection (Mock) vs. PS1KD and PSdKO. (A,C,E) General profile of the OCR with vertical lines indicating the time point at which the different compounds have been added: a. Oligomycin (CV inhibitor) b. FCCP (ΔΨ uncoupler) c. Rotenone (CI inhibitor) and antimycin A (CIII inhibitor). Values (means ± sem) are given in pmol O₂/min/μg protein (min N = 3). (B,D,F) The basal respiration, the coupling ratio and the spare respiratory capacity were calculated according to the Cell Mito Stress Test kit's recommended protocol. Values (means ± sem) are given as percentage of signal measured in the respective control cells (Ct or Mock) (min N = 3). ANOVA and Tukey's multiple comparison test.