Journal: Neural Regeneration Research

Article Title: Rapamycin reverses ferroptosis by increasing autophagy in MPTP/MPP + -induced models of Parkinson’s disease

doi: 10.4103/1673-5374.371381

Figure Lengend Snippet: Rapa protects against dopaminergic neuron loss in the SN of MPTP-treated mice. (A) Western blotting analysis of TH and α-synuclein protein levels. (B) TH-positive cells were stained by immunocytochemistry and observed through a fluorescence microscope. Black arrows indicate TH-positive cells. Scale bars: 500 μm (upper) and 100 μm (lower). Lower panels are enlarged images of the boxes in the upper panels. (C) Number of TH-positive cells. The experiments were performed in triplicate. Data are expressed as the mean ± SEM ( n = 6–8 per group). * P < 0.05, ** P < 0.01 (one-way analysis of variance followed by Tukey’s post hoc test). GAPDH: Glyceraldehyde 3-phosphate dehydrogenase; MPTP: 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine; Rapa: rapamycin; RSL3: methyl (1S,3R)-2-(2-chloroacetyl)-1-(4-methoxycarbonylphenyl)-1,3,4,9-tetrahyyridoindole-3-carboxylate; SN: substantia nigra; TH: tyrosine hydroxylase.

Article Snippet: Membranes were blocked with 5% nonfat milk for 1 hour at room temperature, and then incubated overnight at 4°C with primary antibodies: rat anti-GAPDH (glyceraldehyde 3-phosphate dehydrogenase; monoclonal, 1:1000; Cell Signaling Technology, Boston, USA, Cat# 2118, RRID: AB_561053), rabbit anti-TH (polyclonal, 1:1000; Cell Signaling Technology, Cat# 2792, RRID: AB_2303165), rabbit anti-α-synuclein (monoclonal, 1:1000; Cell Signaling Technology, Cat# 23706, RRID: AB_2798868), rabbit anti-SLC7A11 (polyclonal, 1:1000; Cell Signaling Technology, Cat# 98051, RRID: AB_2800296), rabbit anti-GPX4 (polyclonal, 1:1000; Cell Signaling Technology, Cat# 52455, RRID: AB_2924984), rabbit anti-p62 (monoclonal, 1:1000; Cell Signaling Technology Cat# 8025, RRID: AB_10859911), and rabbit anti-LC3 B (monoclonal, 1:1000; Cell Signaling Technology Cat# 3868, RRID: AB_2137707).

Techniques: Western Blot, Staining, Immunocytochemistry, Fluorescence, Microscopy

Journal: Neural Regeneration Research

Article Title: Rapamycin reverses ferroptosis by increasing autophagy in MPTP/MPP + -induced models of Parkinson’s disease

doi: 10.4103/1673-5374.371381

Figure Lengend Snippet: Rapa inhibits ferroptosis in MPTP-treated mice. (A) Western blotting analysis of glutathione peroxidase 4 (GPX4) and recombinant solute carrier family 7, member 11 (SLC7A11). The concentrations of glutathione (GSH; B), malondialdehyde (MDA; C) and reactive oxygen species (ROS; D) in the supernatant of the brain after treatment. The experiments were performed in triplicate. Data are expressed as the mean ± SEM ( n = 6–8 per group). * P < 0.05, ** P < 0.01 (one-way analysis of variance followed by Tukey’s post hoc test). GAPDH: Glyceraldehyde 3-phosphate dehydrogenase; MPTP: 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine; Rapa: rapamycin; RSL3: methyl (1S,3R)-2-(2-chloroacetyl)-1-(4-methoxycarbonylphenyl)-1,3,4,9-tetrahyyridoindole-3-carboxylate.

Article Snippet: Membranes were blocked with 5% nonfat milk for 1 hour at room temperature, and then incubated overnight at 4°C with primary antibodies: rat anti-GAPDH (glyceraldehyde 3-phosphate dehydrogenase; monoclonal, 1:1000; Cell Signaling Technology, Boston, USA, Cat# 2118, RRID: AB_561053), rabbit anti-TH (polyclonal, 1:1000; Cell Signaling Technology, Cat# 2792, RRID: AB_2303165), rabbit anti-α-synuclein (monoclonal, 1:1000; Cell Signaling Technology, Cat# 23706, RRID: AB_2798868), rabbit anti-SLC7A11 (polyclonal, 1:1000; Cell Signaling Technology, Cat# 98051, RRID: AB_2800296), rabbit anti-GPX4 (polyclonal, 1:1000; Cell Signaling Technology, Cat# 52455, RRID: AB_2924984), rabbit anti-p62 (monoclonal, 1:1000; Cell Signaling Technology Cat# 8025, RRID: AB_10859911), and rabbit anti-LC3 B (monoclonal, 1:1000; Cell Signaling Technology Cat# 3868, RRID: AB_2137707).

Techniques: Western Blot, Recombinant