Fig. 8: VPS35 levels in ALS iPSCs-derived MNs and postmortem biopsies.
From: Retromer stabilization results in neuroprotection in a model of Amyotrophic Lateral Sclerosis
a, b IHC for VPS35 in alpha-MNs in representative sections from ventral horns of SCs biopsies from a non-neurological control (a) and an ALS patient (b). c, d, IHC for VPS26 in alpha-MNs sampled in adjacent sections from a non-neurological control (c) and an ALS patient (d). Analyses were done on four non-neurological control biopsies and five ALS biopsies examined over three independent experiments. e Maximum projections of representative confocal stacks (three independent experiments) for VPS35 and ISLET1 in representative cultures of iPSC-derived MNs from a healthy volunteer (#8) and an ALS patient (#13SOD1Leu144Phe). A representative WB for VPS35 and β-Actin in protein extracts from iPSCs-derived MN cultures (#8 SOD1Asn65Ser, #13SOD1Leu144Phe, #27 SOD1Asp97Asn and age- and sex-matched healthy volunteers #2, #4, and #8) is shown in f. Quantifications derived from n = 3 controls (#2, #4, and #8) and from n = 3 ALS (#8, #13, and #27), means (±SD) are examined from two independent experiments (p = 0.005). g Representative cross-sectional analysis of confocal stacks (three independent experiments) for VPS35 and ISLET1 in cultured iPSCs-derived MNs from an healthy volunteer (#8) and from an ALS patient (#27 SOD1Asp97Asn) with or without compound 2a (10 µM for 6 days). h Representative WB for VPS35 and β-Actin in iPSCs-derived MNs of an ALS patient (#27) treated with vehicle or with compound 2a (10 µM for 6 days). Quantifications (means ± SD) derived from n = 3 wells/treatment and data are examined from one experiment (p = 0.017). Two-tailed Student’s t-test was used to determine the statistical significance in f and h. **p < 0.01. Scale bar: 150 µm in d, 25 µm in e, 15 µm in g.
