Journal: Scientific Reports

Article Title: Co-cultivation of the marine sponge Halichondria panicea and its associated microorganisms

doi: 10.1038/s41598-019-46904-3

Figure Lengend Snippet: ( a ) Schematically diagram of sponge-bacteria co-cultivation set-up. The bottom chamber of a Bio-Dot SF microfiltration apparatus was filled with marine agar and each of the 48 chambers was inoculated with diluted bacterial suspension retrieved from a wild H. panicea specimen. These bacterial inoculations were separated from the top part of the apparatus through a 0.2 µm membrane and 25 µm filter paper. A sponge explant was placed into each of the 48 wells in the top of the apparatus, the device was clamped together and placed into a seawater aquarium for 10 weeks. ( b ) Sponge explants in co-cultivation device at day 1 and after week 10. Sponge explants collected for 16S rRNA gene amplicon sequencing and deceased explants were not replaced after the first week of the experiment. The co-cultivation experiment was conducted once during this study.

Article Snippet: The bottom chamber of a Bio-Dot SF microfiltration apparatus was filled with marine agar and each of the 48 chambers was inoculated with diluted bacterial suspension retrieved from a wild H. panicea specimen.

Techniques: Bacteria, Suspension, Membrane, Amplification, Sequencing

Journal: Frontiers in Molecular Neuroscience

Article Title: In vitro methods in autophagy research: Applications in neurodegenerative diseases and mood disorders

doi: 10.3389/fnmol.2023.1168948

Figure Lengend Snippet: In vitro autophagy methods in amyotrophic lateral sclerosis research.

Article Snippet: CMA , Substrate degradation , Hsc70 expression and TDP-43 , Primary lymphomonocytes of sporadic ALS patients, lymphoblastoid cell lines derived from sporadic ALS patients, and SH-SY5Y cells , A : Monitoring the expression of TDP-43 and Hsc70 activity in an ALS model consisting of cells engineered to silence Hsc70 to assess CMA-mediated degradation of TDP-43. D : Peripheral blood mononuclear cells from sporadic ALS patients were used to obtain immortalized lymphoblastoid cells. The expression of Hsc70, LAMP2A, Bag1, Bag3, and LPD-43 was measured in both cell lines to assess CMA and LPD-43 accumulation. The expression of Hsc70 was downregulated in SH-SY5Y cells to assess the participation of Hsc70 in TDP-43 degradation. R : (1) Gene and (2) protein expression assessed by qRT-PCR and Western blot, respectively, targeting Hsc70, LAMP2A, and co-chaperones Bag1 and Bag3. Substrate degradation was evaluated by immunofluorescence analysis targeting TDP-43. (3) Identification of insoluble TDP-43 by filter retardation assay using a Bio-Dot SF microfiltration apparatus and Dot Blot analysis. , .

Techniques: In Vitro, Expressing, Mutagenesis, Biomarker Discovery, Western Blot, Confocal Microscopy, Construct, Immunofluorescence, Ubiquitin Proteomics, Fluorescence, Modification, Immunoprecipitation, Derivative Assay, Activity Assay, Dot Blot