Journal: Journal of Neurochemistry

Article Title: ATG14 ‐Mediated SNARE Complex Activation Promotes ΔFosB Degradation to Ameliorate Levodopa‐Induced Dyskinesia

doi: 10.1111/jnc.70431

Figure Lengend Snippet: ATG14 overexpression improves SNARE formation, autophagy function and the clearance of ΔFosB in LID rats. (A) The protein levels of ATG14, STX17, SNAP29, VAMP8, LC3, p62, and ΔFosB were detected by western blotting in the striatum of AAV‐GFP and AAV‐ATG14 group rats ( n = 3). GAPDH as an internal reference. (B) The interaction levels of ATG14 and SNARE complex (namely STX17, SNAP29, and VAMP8) were detected by co‐immunoprecipitation in the striatum of each group ( n = 1). (C) The ΔFosB mRNA levels in the striatum of each group were detected by real‐time quantitative polymerase chain reaction ( n = 3). (D, G, H, I) The representative immunofluorescence images of ATG14 and STX17 co‐localization, p62 and LC3‐II co‐localization, LAMP2 and LC3‐II co‐localization as well as p62 and ΔFosB co‐localization in the striatum of each group ( n = 3; bar = 50 μm). (E, F) The representative immunohistochemical images of p62 and ΔFosB in the striatum of each group ( n = 3; bar = 50 μm). (J) The representative transmission electron microscopy images of autophagic vacuoles and autolysosomes in each group ( n = 3; N, nucleus; thin red arrows, autolysosomes; bold red arrows, autophagic vacuoles). (Bars represent the mean, error bars represent the SEM; One‐way ANOVA followed by Tukey multiple‐comparisons tests; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; ns, no significant).

Article Snippet: Afterward, we prepared reaction mixtures by combining cDNA, primers, ddH 2 O, Dye 1, and ChamQ SYBR qPCR Master Mix (Cat. No. Q311, Vazyme), and then dispensed the mixtures into PCR plates.

Techniques: Over Expression, Western Blot, Immunoprecipitation, Real-time Polymerase Chain Reaction, Immunofluorescence, Immunohistochemical staining, Transmission Assay, Electron Microscopy

Journal: Journal of Neurochemistry

Article Title: ATG14 ‐Mediated SNARE Complex Activation Promotes ΔFosB Degradation to Ameliorate Levodopa‐Induced Dyskinesia

doi: 10.1111/jnc.70431

Figure Lengend Snippet: Autophagy deficits and ΔFosB accumulation improved by ATG14 overexpression were aggravated by chloroquine in the striatum of LID rats. (A) The protein levels of ATG14, STX17, SNAP29, VAMP8, LC3, p62, and ΔFosB were detected by western blotting in the striatum of AAV‐ATG14+saline and AAV‐ATG14+CQ group rats ( n = 3). GAPDH was used as an internal reference. (B) The ΔFosB mRNA levels in the striatum of each group were detected by real‐time quantitative polymerase chain reaction ( n = 3). (C, D) The representative immunohistochemical images of p62 and ΔFosB in the striatum of each group ( n = 3; bar = 50 μm). (E, F, G, I) The representative immunofluorescence images of ATG14 and STX17 co‐localization, p62 and LC3‐II co‐localization, LAMP2 and LC3‐II co‐localization as well as p62 and ΔFosB co‐localization in the striatum of each group ( n = 3; bar = 50 μm). (H) The representative transmission electron microscopy images of autophagic vacuoles and autolysosomes in each group ( n = 3; N, nucleus; thin red arrows, autolysosomes; bold red arrows, autophagic vacuoles). (Bars represent the mean, error bars represent the SEM; One‐way ANOVA followed by Tukey multiple‐comparisons tests; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; ns, no significant).

Article Snippet: Afterward, we prepared reaction mixtures by combining cDNA, primers, ddH 2 O, Dye 1, and ChamQ SYBR qPCR Master Mix (Cat. No. Q311, Vazyme), and then dispensed the mixtures into PCR plates.

Techniques: Over Expression, Western Blot, Saline, Real-time Polymerase Chain Reaction, Immunohistochemical staining, Immunofluorescence, Transmission Assay, Electron Microscopy