Journal: International Journal of Molecular Sciences

Article Title: Identification of KHS-101 as a Transcription Factor EB Activator to Promote α-Synuclein Degradation

doi: 10.3390/ijms27020905

Figure Lengend Snippet: KHS-101 increases α-synuclein (A53T) degradation via autophagic–lysosomal system. ( A , B ) Representative immunoblot ( A ) and quantification ( B ) of α-synuclein levels. SH-SY5Y cells were treated with KHS-101 (1 μM, 2 μM, 4 μM, 6 μM) and doxycycline (1 μg/mL) for 48 h. ( C , D ) Representative immunofluorescence images ( C ) and quantification ( D ) of α-synuclein levels in SH-SY5Y cells obtained after 48 h treatment with KHS-101 (6 μM) and doxycycline (1 μg/mL). Scale bars represent 10 μm. ( E , F ) α-synuclein degradation by KHS-101 was abolished in the presence of CQ. SH-SY5Y cells were treated with KHS-101 (6 μM) and doxycycline (1 μg/mL) for 48 h with or without CQ (5 μM). All data are expressed as mean ± SEM (** p < 0.01; *** p < 0.001; **** p < 0.0001).

Article Snippet: The cDNA sequences of TFEB and α-synuclein (A53T) were purchased from Sino Biological (Beijing, China).

Techniques: Western Blot, Immunofluorescence

Journal: bioRxiv

Article Title: Gangliosides Modulate the Secretion of Extracellular Vesicles and Their Misfolded Protein Cargo

doi: 10.1101/2025.04.03.647108

Figure Lengend Snippet: a . Particle size distribution profile of EVs secreted by N2a cells treated with GM1 compared to untreated cells, as measured by NTA. EV concentration was normalized to a common scale. The table reports the mean particle diameter ± SD and p -values calculated by two-tailed paired t -test (n=3 independent experiments). b. Schematic of the ExoView tetraspanin-capture chip and the method used to capture and profile EVs. EVs are captured onto chips by anti-tetraspanin antibodies printed onto the chips. EVs are then labeled with fluorescent anti-tetraspanin antibodies and imaged for tetraspanin profiling and sizing by SP-IRIS. c. Particle size distribution profile of EVs secreted by human i 3 Neurons, human fibroblasts and HeLa cells treated with GM1compared to untreated cells, as detected by SP-IRIS. EV counts were normalized to a common scale. The table inserts report the mean particle diameter ± SD and p -values obtained by two-tailed paired t -test (n=2-3 independent experiments). d. Tetraspanin profile of EVs secreted by human i 3 Neurons, human fibroblasts and HeLa cells treated with GM1 compared to untreated cells. Colored bars show the proportion of EVs bearing the indicated tetraspanin combinations. * p <0.05 by two-way ANOVA of probit-transformed data with Šídák’s post-hoc test (n=2-3 independent experiments). e. Representative dot blot of cholera toxin B binding to quantify GM1 in cell lysates and EVs from N2a cells treated with 50 µM GM1 for 6 h, compared to untreated cells. GM1 was washed off and EVs were collected for 16 h and isolated by SEC. The graph shows the densitometric analysis of cholera toxin B binding to EV fractions, normalized to DiI EV fluorescence. Bars are mean ± SD. * p <0.05 by two-tailed paired t -test (n=3 independent experiments).

Article Snippet: N2a cells were stably transfected with the cDNA for mHTT Exon 1 (containing 97 glutamines) tagged with enhanced green fluorescent protein (HTT-Exon1 (97Q) EGFP) kindly donated by Dr. Janice Braun (University of Calgary, Canada), or α-synuclein (A53T) tagged with enhanced green fluorescent protein purchased from Addgene (plasmid #40823).

Techniques: Concentration Assay, Two Tailed Test, Labeling, Transformation Assay, Dot Blot, Binding Assay, Isolation, Fluorescence