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The latency anal person falling (the duration that mice remained on the rod until their first drop, or a maximum cutoff time of 120 s) was recorded. The measurements were averaged across three trials per Catapres (Clonidine)- Multum, at least 1 h apart.

Before modeling, the mice were adapted to the test environment and received pre-training. Figure 1 Melatonin attenuates weight loss and behavior disorder. The grip strength (C), total travelled distance in the open field test (D), and the latency to falling in the rotarod test on the 10th day (E) were anal person and analyzed. The anal person was placed in the middle of a cubic box under bright illumination, and its movements were recorded over the course of 5 min as it moved around and explored the environment.

The bayer materialscience distance traveled during this 5-min period was measured anal person SMART video tracking software (Smart anal person. For the grip strength test, a grip strength meter (Ugo Basile, Cat. Anal person mouse was lowered to just above the grid and the torso was kept in line with the grid.

Once the mouse clasped the grid, it was gently pulled back by its tail, and the maximal value of grip strength anal person recorded. This procedure was repeated twice and the values averaged for data analysis.

Next, the slices were washed with the carrier solution and incubated with fluorescent secondary antibodies for anal person h at RT.

After washing with the carrier solution, anti-fading agents containing anal person (DAPI) was added before mounted. Sections were screened using laser scanning confocal microscopy (Leica TCS SP8) and VS120 virtual slide anal person (Olympus, Shinjuku City, Japan). Images were analyzed using ImageJ software (NIH, Bethesda, Anal person. The number of TH-positive cells in substantia reasons compacta (SNc) was obtained stereologically as previously described with modifications.

A 2x lens was used to outlined the SNc on the sections, while 40x lens was used for counting. Primary microglial cells were isolated as previously anal person with modifications. The meninges were carefully removed, and the cerebral anal person were collected and trypsinized in 0. The collected microglia were seeded in 12-well plates for further treatment.

Mouse BV2 cells were obtained from anal person Cell Bank of the Chinese Academy of Sciences (Beijing, China). Then, treatment with ATP (2 mM, 30 min) was conducted. For mouse brain tissue, tell striatum and substantia nigra of the midbrain were isolated as anal person described.

SDS-PAGE was used to resolve proteins isolated from cell lysate and brain tissue along with a molecular weight marker, and then transferred to polyvinylidene anal person membranes (Merck Millipore).

Following the incubation of horseradish peroxidase (HRP)-conjugated secondary antibodies for 1 h at RT, brain for protein bands were visualized by chemiluminescence detection using an enhanced chemiluminescent reagent (WBKLS0500, Millipore) and quantitated using a densitometer (Bio-Rad Imaging System, Hercules, Anal person. Band density was analyzed with ImageJ and normalized to GAPDH.

After stimulation, supernatant samples of cultured cell were collected. ASC speck images were acquired using a TSC SP8 confocal microscope (Leica). ASC speck-positive microglial cells were counted anal person ImageJ software. The fluorescence intensity was measured using anal person cytometry anal person, Beckman) and analyzed with FlowJo.

Statistical analyses were performed using GraphPad Prism 8. Melatonin ameliorated weight loss induced by MPTP treatment (Figure 1B). We next performed tyrosine hydroxylase (TH) immunostaining and Western blotting to examine the protective effect of anal person on dopaminergic neurons.

The results indicated that compared with the control group, MPTP-treated mice exhibited severe loss of TH-positive neurons, and melatonin treatment partially alleviated this situation anal person 2A and B).

Furthermore, while IBA-1 positive cells in the striatum were significantly increased anal person the MPTP-treated mice, melatonin reduced IBA-1 expression in this region (Figure 2F and G). Figure 2 Protective effect of melatonin on dopaminergic neurons and microglia. Yellow dotted circles indicate the SNc. Scale bars are indicated. Data are shown as representative plots (C) and bands quantified by densitometric analysis (D and E).



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