Journal: Journal of Cancer Research and Clinical Oncology

Article Title: Effect of human bone marrow mesenchymal stem cell-derived microvesicles on the apoptosis of the multiple myeloma cell line U266

doi: 10.1007/s00432-024-05822-2

Figure Lengend Snippet: Western blotting assay for the verification of the BMSC-MVs. CD63, CD73, and CD81 with approximately 50, 75, and 26 kDa protein bands, respectively, were detected

Article Snippet: Next, the membranes were separately incubated with the following primary antibodies: rabbit polyclonal anti-human CD63 antibodies (cat. No. 11,271-T16; Sino Biological, China), rabbit monoclonal anti-human CD73 antibodies (cat No. ab124725; Abcam, MA, United States), and rabbit polyclonal anti-human CD81 antibodies (cat No. ab155760; Abcam, MA, United States) as per the suppliers’ instructions.

Techniques: Western Blot

Journal: Biochemical and biophysical research communications

Article Title: Choline kinase alpha regulates autophagy-associated exosome release to promote glioma cell progression.

doi: 10.1016/j.bbrc.2024.151269

Figure Lengend Snippet: Fig. 3. CHKA mediates exosomes secretion in glioma cells. A-C Western blot (WB) analysis of exosome marker proteins CD63 and TSG101 in exosomes isolated from the supernatant of glioma cells in shCHKA and shNC group along with the negative control protein Calnexin. D Exosome protein concentration in the shCHKA and shNC groups of glioma cells using the BCA assay. E Electron microscopy was performed on shCHKA and shNC cells in glioma cells (Scale bars = 1 μm). F The number of multivesicular bodies (MVBs) per glioma cell profile. G The number of intraluminal vesicles (ILVs) per MVB profile in glioma cells. *P < 0.05, **P < 0.01, ***P < 0.001. shCHKA: CHKA knockdown group, shNC: con trol group.

Article Snippet: Antibodies used in the Western blotting process included: rabbit antihuman CHKA polyclonal antibody (abcam, USA), rabbit anti-human GAPDH polyclonal antibody (Affinity, China), rabbit anti-human CD63 polyclonal antibody (Proteintech, China), rabbit anti-human TSG101 polyclonal antibody(Proteintech, China), rabbit anti-human calnexin polyclonal antibody(Proteintech, China), rabbit anti-human LC3B polyclonal antibody(Sigma, USA), and rabbit anti-human P62/SQSTM1 polyclonal antibody(Sigma, USA).

Techniques: Western Blot, Marker, Isolation, Negative Control, Protein Concentration, BIA-KA, Electron Microscopy, Knockdown

Journal: Biochemical and biophysical research communications

Article Title: Choline kinase alpha regulates autophagy-associated exosome release to promote glioma cell progression.

doi: 10.1016/j.bbrc.2024.151269

Figure Lengend Snippet: Fig. 4. CHKA knockdown Inhibited Proliferation, Migration and invasion of glioma Cells by Blocking Exosome Secretion. A Western blot (WB) analysis of exosome marker proteins CD63 and TSG101 in exosomes isolated from the supernatant of glioma cells in DMSO and GW4869 group. (BC) The proliferation of glioma cells U87MG and U251 in DMSO and GW4869 group, the migration ability of glioma cells U87MG (DE) and U251 (DF) in DMSO and GW4869 group (Scale bars = 200 μm). GH The invasion ability of glioma cells U87MG and U251 in DMSO and GW4869 group(Scale bars = 100 μm). *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Antibodies used in the Western blotting process included: rabbit antihuman CHKA polyclonal antibody (abcam, USA), rabbit anti-human GAPDH polyclonal antibody (Affinity, China), rabbit anti-human CD63 polyclonal antibody (Proteintech, China), rabbit anti-human TSG101 polyclonal antibody(Proteintech, China), rabbit anti-human calnexin polyclonal antibody(Proteintech, China), rabbit anti-human LC3B polyclonal antibody(Sigma, USA), and rabbit anti-human P62/SQSTM1 polyclonal antibody(Sigma, USA).

Techniques: Knockdown, Migration, Blocking Assay, Western Blot, Marker, Isolation

Journal: Biochemical and biophysical research communications

Article Title: Choline kinase alpha regulates autophagy-associated exosome release to promote glioma cell progression.

doi: 10.1016/j.bbrc.2024.151269

Figure Lengend Snippet: Fig. 6. CHKA attenuates exosomes release through Autophagy-Mediated mechanism AB Western blot (WB) analysis of exosome marker proteins CD63 and TSG101 in exosomes isolated from the supernatant of U87MG cells in shNC + DMSO, shCHKA + DMSO and shCHKA + Baf-A1 group. AC Western blot (WB) analysis of exosome marker proteins CD63 and TSG101 in exosomes isolated from the supernatant of U251 cells in shNC + DMSO, shCHKA + DMSO and shCHKA + Baf-A1 group. D Exosome protein concentration in the s shNC + DMSO, shCHKA + DMSO and shCHKA + Baf-A1 groups of glioma cells using the BCA assay. **P < 0.01, ***P < 0.001. Baf-A1: Bafilomycin A1.

Article Snippet: Antibodies used in the Western blotting process included: rabbit antihuman CHKA polyclonal antibody (abcam, USA), rabbit anti-human GAPDH polyclonal antibody (Affinity, China), rabbit anti-human CD63 polyclonal antibody (Proteintech, China), rabbit anti-human TSG101 polyclonal antibody(Proteintech, China), rabbit anti-human calnexin polyclonal antibody(Proteintech, China), rabbit anti-human LC3B polyclonal antibody(Sigma, USA), and rabbit anti-human P62/SQSTM1 polyclonal antibody(Sigma, USA).

Techniques: Western Blot, Marker, Isolation, Protein Concentration, BIA-KA

Journal: Annals of Medicine

Article Title: The expression and clinical significance of serum exosomal-long non-coding RNA DLEU1 in patients with cervical cancer

doi: 10.1080/07853890.2024.2442537

Figure Lengend Snippet: The expression of serum exosomal DLEU1 in patients with CC. (A) The morphological characteristics of exosomes were observed under TEM; (B) the sizes of exosomes were detected using NTA; (C) the expression of TSG101, CD63 and calnexin was detected by Western blotting; D and E: DLEU1 was highly expressed in CC tissues; (F) DLEU1 was highly expressed in serum exosomes of patients with CC. ns: no significant difference. *** p < .001.

Article Snippet: The membranes were blocked with 1% bovine serum albumin overnight, incubated with monoclonal rabbit anti-human antibodies against CD63 (1:1000, Proteintech, Wuhan, China), TSG101 (1:5000, Proteintech, Wuhan, China) and calnexin (1:1000, Abcam, Cambridge, UK) for 2 h, washed with TBST, incubated with a goat anti-rabbit secondary antibody at 37 °C for an hour, and then immersed in 1 mL of the developing solution in the dark for approximately 10 min.

Techniques: Expressing, Western Blot

Journal: Experimental and Therapeutic Medicine

Article Title: Exosomes from Ub‑HBcAg‑overexpressing dendritic cells induce T‑lymphocyte differentiation and enhance cytotoxic T‑lymphocyte activity

doi: 10.3892/etm.2023.11866

Figure Lengend Snippet: Isolation and identification of Dexs-Ub-HBcAg. (A) Transmission electron microscopy of the mDexs ultrastructure. Scale bar, 200 nm. (B) Expression of the positive exosomal markers CD9, CD63 and TSG101, and of HBcAg was examined via the western blot analysis of Dexs lysates. (C) Size dispersion profile of mDexs evaluated using ZetaVIEW ® nanoparticle tracking analysis, indicating a size peak of 123.8 nm. Dexs, dendritic cell-derived exosomes; HBcAg, hepatitis B virus core antigen; Dexs-Ub-HBcAg, Dexs loaded with ubiquitinated HBcAg; Con, control; TSG101, tumor susceptibility gene 101.

Article Snippet: Subsequently, the membrane was blocked with 5% non-fat milk at room temperature for 1 h. The primary antibodies used were rabbit anti-mouse CD63 (1:500; cat. no. ab216130; Abcam), anti-CD9 (1:500; cat. no. ab92726; Abcam) and tumor susceptibility gene 101 (TSG101; 1:500; cat. no. 102286-T38; SinoBiological) monoclonal antibodies.

Techniques: Isolation, Transmission Assay, Electron Microscopy, Expressing, Western Blot, Derivative Assay