Abbreviated schematic presentation of sphingolipid metabolism.

<p>Gal, galactosyl; Glu, glucosyl; Lac, lactosyl; S-1-P, sphingosine-1-phosphate. Red indicates lowered levels and blue indicates increased levels observed in our study.</p

Patient demographic and neuropathologic features of control, Lewy body disease with intermediate Alzheimer’s pathology (LBD-I-AD), Lewy body disease with Alzheimer’s pathology (LBD-AD), and Parkinson’s disease (PD) subjects.

<p>PMI (post-mortem interval).</p

Exact masses and MS² data for reported lipids.

<p>Exact masses and MS² data for reported lipids.</p

Abbreviated schematic presentation of DAG metabolism.

<p>LPA, lysophosphatidic acid; PC, phosphatidylcholine; PG, phosphatidylglycerol; PS, phosphatidylserine; PLC, phospholipase C; PLD, phospholipase D; SMS, sphingomyelin synthase; DGK, diacylglycerol kinase. Red indicates lowered levels and blue indicates increased levels observed in our study.</p

Frontal cortex levels of diacylglycerols (DAG) and lysophosphatidic acid 16:0 (LPA) in control, Lewy body disease with intermediate Alzheimer’s disease (LBD-I-AD), Lewy body disease with Alzheimer’s disease (LBD-AD), and Parkinson’s disease (PD) tissues.

<p>R = ratio of the peak area of the endogenous lipid to the peak area of the internal standard (mean ±SEM). Analysis of PD subgroups demonstrated that DAGs were augmented in the cortex of PD subjects with no neocortical pathology (PD-1, N = 5), subjects with sparse neocortical neuritic plaques (PD-2, N = 5), and subjects with moderate to frequent neocortical neuritic plaques (PD-3, N = 5). *, p < 0.01 vs. controls; #, p < 0.05 for PD-2 vs. PD3.</p

Decreased frontal cortex levels of 20:5-containing phosphatidylcholines (PtdC) in Lewy body disease with intermediate Alzheimer’s disease (LBD-I-AD), Lewy body disease with Alzheimer’s disease (LBD-AD), and Parkinson’s disease (PD) tissues.

<p>R = ratio of the peak area of the endogenous PtdC to the peak area of the internal standard (mean ±SEM). *, p < 0.01; #, p < 0.05 vs controls.</p

Frontal cortex levels of plasmalogens in control, Lewy body disease with intermediate Alzheimer’s disease (LBD-I-AD), Lewy body disease with Alzheimer’s disease (LBD-AD), and Parkinson’s disease (PD) tissues.

<p>PlsC, choline plasmalogens; PlsE, ethanolamine plasmalogen. R = ratio of the peak area of the endogenous plasmalogen to the peak area of the internal standard (mean ±SEM). *, p < 0.05 vs controls.</p

Bafilomycin A1/cediranib efficacy and bafilomycin A1 effects on LC3 and Akt.

<p>A) Results from MTS cell viability assays showing mean 4C8 cell viability in response to treatment, normalized to untreated samples. Assays were performed after cells were exposed to cediranib and/or bafilomycin A1 treatment for 72 hours, under normoxic or hypoxic (0.5% O₂) conditions. Results are mean ± SEM of three independent experiments, performed in triplicate. B) Representative LC3 western blots obtained from 4C8 cells grown under normoxic or hypoxic (0.5% O₂) conditions for 10 hrs while untreated or exposed to quinacrine (0.8 µM) or bafilomycin A1 (5 nM). C) Representative basal Akt and phospho-Akt western blots obtained from 4C8 cells grown under hypoxic (0.5% O₂) conditions for 24 hrs while untreated or exposed to bafilomycin A1 (5 nM).</p

Cediranib and quinacrine inhibit Akt activation.

<p>Western blot analysis of basal Akt and phospho-Akt levels in 4C8 cultures grown under hypoxic (0.5% O₂) conditions for 24 hrs while untreated or exposed to cediranib (2.5 µM), quinacrine (2.5 µM), or combined cediranib/quinacrine.</p

The effect of hypoxia and cediranib/quinacrine on LC3.

<p>A) Representative LC3 western blots obtained from 4C8 cells grown under normoxic or hypoxic (0.5% O₂) conditions for 10 hrs while untreated or exposed to cediranib (3 µM), quinacrine (0.8 µM), or combined cediranib/quinacrine. B) RFP-LC3 expressing 4C8 cells were visualized using Zeiss LSM 780 laser scanning confocal microscope after incubation under hypoxic conditions for 24 hr while untreated or exposed to cediranib (2.5 µM), quinacrine (2.5 µM), or combined cediranib/quinacrine.</p