Journal: Neuro-Oncology Advances

Article Title: Plasma extracellular vesicle sampling from glioblastoma demonstrates a small RNA signature indicative of disease and identifies lncRNA RPPH1 as a biomarker

doi: 10.1093/noajnl/vdaf273

Figure Lengend Snippet: (A) Western blot analysis demonstrating EVs isolated from pooled GBM-EVs utilizing Vn96 peptide affinity have canonical markers of EVs including Hsc70, CD63, FLOT1 and CD9. The non-EV marker GRP94 is detected at low levels. HEK 293T total cell lysate (HEK-CL) was used as a positive control. Distribution of multi-annotated, uniquely annotated and not annotated RNA reads upon sequencing control and GBM samples (B); upset plot illustrating the distribution and frequency of uniquely annotated RNA sequences and multi-annotated sequences in plasma stratified by type of annotated small RNA. The columns are sorted according to a decreasing number of uniquely annotated RNA, double-annotated RNA and multi-annotated RNA with 3 or more different annotations (C).

Article Snippet: Primary antibodies were prepared in TBST with 5% skim milk: Hsc-70 (1:200, Santa Cruz Biotechnology Cat# SC-7298, RRID: AB_627761), CD63 (1:200, Santa Cruz Biotechnology Cat# SC-5275, RRID: AB_627877), FLOT1 (1:1000, Cell Signaling Technology, Cat #18634, RRID: AB_2773040), CD9 (1:200, Santa Cruz Biotechnology Cat# SC-59140, RRID: AB_1120766) and GRP94 (1:1000, New England Biolabs, Cat#2104S, RRID: AB_823506).

Techniques: Western Blot, Isolation, Marker, Positive Control, Sequencing, Control, Clinical Proteomics