Journal: PLoS Pathogens

Article Title: Probing the spatiotemporal patterns of HBV multiplication reveals novel features of its subcellular processes

doi: 10.1371/journal.ppat.1009838

Figure Lengend Snippet: (A) HepAD38 cells were fixed and processed for detecting HBV (-) DNA, HBsAg and MVBs specific marker CD63. Specific enlargement of the area within the white-outlined box, colocalization of (-) DNA, HBsAg and CD63 were presented by white arrows. Scale bar, 4 μm. (B) Quantification of images in DMSO-treated cells and analyzing the relationship between the mean volume of CD63 + vesicles and (-) DNA puncta in each cell. (C, D) Mean volume and puncta of CD63 + vesicles in a single cell level were measured by Huygens. Analyzing (-) DNA and CD63 + positive (E), HBsAg and CD63 + positive (F) colocalization percent per cell within 3 μm. More than 15 cells per group was counted. ****P < 0.0001 (Mann-Whitney U-test).

Article Snippet: Monoclonal antibodies against acetyl-Histone H3 (Lys27) (D5E4), RNA polymerase II (8WG16), Smc6 (M01), puromycin (4G11) and CD63 (H5C6) were purchased from Cell-Signaling Technology (8173, USA), Abcam (ab817, UK), Abgent (AT3956a, USA), Sigma-Aldrich (MABE342, MO) and BD Biosciences (556019, CA), respectively.

Techniques: Marker, MANN-WHITNEY

Journal: Molecular Therapy. Nucleic Acids

Article Title: A novel function and mechanism of ischemia-induced retinal astrocyte-derived exosomes for RGC apoptosis of ischemic retinopathy

doi: 10.1016/j.omtn.2024.102209

Figure Lengend Snippet: Antibodies used in this study

Article Snippet: Anti-CD63 rabbit pAb , System Biosciences, Cat#. EXOAB-CD63A-1.

Techniques:

Journal: BBA Advances

Article Title: Valosin-containing protein (VCP), a component of tumor-derived extracellular vesicles, impairs the barrier integrity of brain microvascular endothelial cells

doi: 10.1016/j.bbadva.2024.100130

Figure Lengend Snippet: Schematic representation of EV isolation protocol and characterization of isolated EVs from MDA-MB-231 parental (MDA-par) and brain-seeking (MDA-br) cells. (A) Differential centrifugation scheme for obtaining EVs. (B) Nano-Tracking Analysis of small EV from MDA-MB-231-parental and (C) MDA-MB-231-brain variant, evidencing the size distribution as a function of particle concentration. (D) Western blot analysis of calnexin, TSG101, CD63, and CD9 in sEV, Large EV and Cell Lysate (CL). Data are presented as the mean ± SEM of three independent experiments ( n = 3).

Article Snippet: Proteins were detected by incubation with the following primary antibodies (1:1000): Calnexin (CST, #2679), CD63 (CST, #55051), CD9 (CST, #98327), TSG101 (CST, #72313), β-actin (Sigma-Aldrich, #A5316), diluted in blocking solution (TBST 1 % in BSA 5 %) for 16h at 4 °C.

Techniques: Isolation, Centrifugation, Variant Assay, Concentration Assay, Western Blot

Journal: Scientific Reports

Article Title: Heterogeneity of extracellular vesicles in porcine myoblasts regulates adipocyte differentiation

doi: 10.1038/s41598-024-77110-5

Figure Lengend Snippet: Skeletal muscle stem cell-derived small extracellular vesicles inhibit porcine adipose precursor cell viability. ( a ) Transmission electron microscopy of sEV derived from porcine skeletal muscle stem cells demonstrated (Note: Blue arrows indicate small extracellular vesicles, scale bar = 200 nm). ( b ) Nanoparticle tracking analysis was used to determine the size distribution and concentration of sEV. ( c ) Western Blot was performed to detect sEV marker proteins Alix, TSG101, CD9, and CD63. ( d ) sEV-treated porcine primary adipocytes were assayed for cell viability by MTT assay (n = 6). Values are presented as means ± SEM, * P < 0.05, and ** P < 0.01, according to the non-paired Student’s t -test or one-way ANOVA between individual groups.

Article Snippet: According to the instructions provided by the manufacturer, the primary antibodies used were rabbit anti-Alix (1:1000, #D262028; Sangon Biotech), rabbit anti-TSG101 (1:2000, #381,538; ZEN BIO), rabbit anti-CD9 (1:1000, #AP68-965-100; Abcepta), rabbit anti-CD63 (1:2000, #D160973; Sangon Biotech), rabbit anti-PPARγ (1:1000, #2443; CST), and rabbit anti-GAPDH (1:5000, #BS65529; Bioworld).

Techniques: Derivative Assay, Transmission Assay, Electron Microscopy, Concentration Assay, Western Blot, Marker, MTT Assay