hnRNP A2/B1 is a choroid plexus and CSF vesicle protein.

<p>A. Nanoparticle tracking analysis of serum free media from human CPE cultures and size frequency distributions. Media contained 559.1 (x10⁹) EVs per mL (N = 3). B. The average size of an EV was 160.3 nm (N = 3). C. Transmission electron micrographs of CPE EVs. Scale bar: 100 nm. D. Immunoblot of the exosome protein ALIX from vesicles isolated from CPEs cultured in the presence or absence of serum. E. The consensus sequence contains features of a recently described exosome miRNA motif recognized by hnRNPA2/B1. F. Multiple sequence alignment of vesicle proteins indicates conservation of miRNA sequence. G. Immunoblot of CPE derived media in the absence of serum or media alone containing serum for ALIX H. Immunoblot of hnRNPA2/B1 from CPE derived media in the absence of serum or media alone containing serum for. I. Immunoblot of hnRNPA2/B1 from CSF EVs demonstrating an age dependent decrease. J. Quantification of immunoblots for hnRNPA2/B1. *: p = 0.0138 Error Bars: SEM.</p

CSF EVs undergo temporal declines.

<p>A. CSF from patients ranging from 1 month to 85 years was collected and subjected to nanoparticle tracking analysis. The average size of EVs was unchanged. B. The average number of EVs was highest in patients less than 2 years and decreased by 10–15 years and 70 years. C. Overlay of size distribution of vesicles from patients less than 2 years, 10-15 years, and greater than 70 years. D–F. Size distribution averages with SEM depicted. G–I. Size distribution averages with area under the curve depicted. *: p<.01 Error Bars: SEM.</p

Next generation sequencing of CSF vesicle RNA content from patients less than 2 years old.

<p>A. Reads aligned to the human genome as a distribution across chromosomes. B. Rank sum distribution of mapped reads as a function of relative abundance (FPKM). C. Frequency distribution of reads in relation to read length. D. Alignment of a novel short RNA, CUFF .1222 to Hg19 chromosome 17 as visualized by integrated genomics viewer. The sequence of Hg19 is seen on the X axis. The chromosomal alignment is shown on the Y axis with red delineating the boundaries of the alignment.</p

Temporal dynamics of CSF vesicle miRNAs.

<p>A. miRNA microarray heat-map of CSF vesicle content from patients less than 2 years or greater than 70 years where red is higher expression and green is lower expression. B. Rank expression of miRNAs based on microarray probe intensity. Red is microarray data from patients less than 2 years and black is from patients greater than 70 years. C. Log(₂) signal of the most abundant miRNAs. D. Hierarchical clustering of miRNA microarrays at different ages. E. Venn diagram of high abundance or (F.) low abundance miRNAs at different ages.</p