Journal: Science Advances

Article Title: Blood-derived APLP1 + extracellular vesicles are potential biomarkers for the early diagnosis of brain diseases

doi: 10.1126/sciadv.ado6894

Figure Lengend Snippet: ( A ) mRNA profiles of APLP1 and L1CAM across different mouse organs. ( B and C ) Relative mRNA expression levels quantified via reverse transcription qPCR, normalized to glyceraldehyde phosphate dehydrogenase ( GAPDH ). Data are presented as the means ± SEM of eight independent experiments, and statistical analysis was performed on 2 Δ C t values using the analysis of variance (ANOVA; post hoc: Tukey). Symbol “***” indicates a value of < 0.001, respectively, compared with the brain. ( D and E ) Western blot representation (D) and immunofluorescence imaging (E) of L1CAM and APLP1 across varied tissues derived from five C57BL/6 mice. ( F ) Use of RNAscope and IHC to detect APLP1 colocalization in mouse cerebral cortex cells. The mRNA of APLP1 is visualized in red, while oligodendrocytes (Olig2), neurons (NeuN), astrocytes (GFAP), or microglial cells (Iba-1) are visualized in green fluorescence. Scale bars, 50 μm. The experiment was performed with three C57BL/6 mice. n.s., not significant; DAPI, 4′,6-diamidino-2-phenylindole.

Article Snippet: The antibodies used for the ELISA were as follows: antihuman CD63 (1:1500 dilution; Ancell, catalog no. 215-820), antihuman CD9 (1:1500 dilution; BD Biosciences, catalog no. 555370; RRID: AB_395772), antihuman CD81 (1:1500 dilution; BD Biosciences, catalog no. 555675; RRID: AB_396028), antihuman L1CAM (1:500 dilution; Cell Signaling Technology, Danvers, MA, USA, catalog no. 89861S; RRID: AB_2800145), antihuman APLP1 (1:500 dilution; Abcam, catalog no. ab94957; RRID: AB_10890629), antimouse immunoglobulin G (IgG) secondary HRP (1:500 dilution; Cell Signaling Technology, catalog no. 7076S; RRID: AB_330924), and antirabbit IgG secondary HRP (1:500 dilution; Cell Signaling Technology, catalog no. 7074S; RRID: AB_2099233).

Techniques: Expressing, Reverse Transcription, Western Blot, Immunofluorescence, Imaging, Derivative Assay, RNAscope, Fluorescence

Journal: Science Advances

Article Title: Blood-derived APLP1 + extracellular vesicles are potential biomarkers for the early diagnosis of brain diseases

doi: 10.1126/sciadv.ado6894

Figure Lengend Snippet: ( A ) Enzyme-linked immunosorbent assay (ELISA)–derived absorbance units representing the levels of EV markers (CD9, CD81, and CD63) and key antigens (EGFR, EGFRviii, L1CAM, and APLP1) within plasma EVs from the healthy group and the GBM patient group. Data represent means ± SEM of three independent experiments. ( B ) Representative immunostaining images contrasting plasma EVs from healthy individuals and patients with GBM ( n = 3). The white arrowheads indicates CD63 + APLP1 + EVs. Scale bars, 5 μm. ( C ) Quantification of CD63 + APLP1 − EVs and CD63 + APLP1 + EVs in each group ( n = 3). ( D ) Fold change ratio of CD63 + APLP1 − EVs and CD63 + APLP1 + EVs between the healthy and GBM groups ( n = 3). Data represent means ± SEM of three independent experiments. Statistical analysis was conducted using the ANOVA test (post hoc: Tukey). ** P < 0.01 and *** P < 0.001, statistical differences.

Article Snippet: The antibodies used for the ELISA were as follows: antihuman CD63 (1:1500 dilution; Ancell, catalog no. 215-820), antihuman CD9 (1:1500 dilution; BD Biosciences, catalog no. 555370; RRID: AB_395772), antihuman CD81 (1:1500 dilution; BD Biosciences, catalog no. 555675; RRID: AB_396028), antihuman L1CAM (1:500 dilution; Cell Signaling Technology, Danvers, MA, USA, catalog no. 89861S; RRID: AB_2800145), antihuman APLP1 (1:500 dilution; Abcam, catalog no. ab94957; RRID: AB_10890629), antimouse immunoglobulin G (IgG) secondary HRP (1:500 dilution; Cell Signaling Technology, catalog no. 7076S; RRID: AB_330924), and antirabbit IgG secondary HRP (1:500 dilution; Cell Signaling Technology, catalog no. 7074S; RRID: AB_2099233).

Techniques: Enzyme-linked Immunosorbent Assay, Derivative Assay, Clinical Proteomics, Immunostaining

Journal: PLoS ONE

Article Title: The Effect of Antenatal Depression and Selective Serotonin Reuptake Inhibitor Treatment on Nerve Growth Factor Signaling in Human Placenta

doi: 10.1371/journal.pone.0116459

Figure Lengend Snippet: Clonality, host species and dilutions of antibodies used in Western blot analysis and immunohistochemistry.

Article Snippet: Primary antibodies were against NGF (catalogue no sc-548), Raf-1 (catalogue no sc-133), p-Raf-1 (Tyr 340/341) (catalogue no sc-16806), RhoA (catalogue no sc-418), ROCK1 (catalogue no sc-5560), ROCK2 (catalogue no sc-1851), all from SantaCruz Biotechnology Inc.,US.

Techniques: Western Blot, Immunohistochemistry

Journal: PLoS ONE

Article Title: The Effect of Antenatal Depression and Selective Serotonin Reuptake Inhibitor Treatment on Nerve Growth Factor Signaling in Human Placenta

doi: 10.1371/journal.pone.0116459

Figure Lengend Snippet: Placental sections stained for NGF, phosphorylated Raf-1 (pRaf-1) and ROCK2 in A) Trophoblasts, B) Endothelial cells and C) Stromal cells.

Article Snippet: Primary antibodies were against NGF (catalogue no sc-548), Raf-1 (catalogue no sc-133), p-Raf-1 (Tyr 340/341) (catalogue no sc-16806), RhoA (catalogue no sc-418), ROCK1 (catalogue no sc-5560), ROCK2 (catalogue no sc-1851), all from SantaCruz Biotechnology Inc.,US.

Techniques: Staining

Journal: eLife

Article Title: P2RY14 cAMP signaling regulates Schwann cell precursor self-renewal, proliferation, and nerve tumor initiation in a mouse model of neurofibromatosis

doi: 10.7554/eLife.73511

Figure Lengend Snippet: ( A ) Microarray heatmap shows P2ry14 receptor expression in p75 + /EGFR + SCP-like tumor-initiating cells derived from human plexiform neurofibroma tumor cells compared to p75 + /EGFR - SCP-like cells. ( B ) Western blot of human Schwann cells and neurofibroma SCP shows the latter has a 1.9-fold increase in P2ry14 protein expression. ( C ) Immunohistochemistry of human neurofibroma shows P2ry14 expression (DAB staining: brown [P2ry14 positive cells] blue [cell nuclei]). ( D ) Representative fluorescence-activated cells sorting (FACS) plot shows live sorted human plexiform neurofibroma tumor cells. ( E ) Representative FACS plot shows human plexiform neurofibroma tumor cells sorted into p75 + /EGFR + SCP-like tumor-initiating cells (pink square). ( F ) Representative FACS plot shows p75 + /EGFR + SCP-like tumor-initiating cells further sorted into p75 + /EGFR + / P2ry14 - (left, purple square) and P75 + /EGFR + / P2ry14 + (right, blue square). ( G ) Photomicrographs of human neurofibromas dissociated using FACS to yield: unsorted, p75 + /EGFR + / P2ry14 - and P75 + /EGFR + / P2ry14 + cells. ( H ) Quantification of unsorted, p75 + /EGFR + / P2ry14 - and P75 + /EGFR + / P2ry14 + cells plated in sphere medium. (n = 3; two-way ANOVA; primary: **p = 0.0057, ****p < 0.0001; secondary: *p = 0.0487; **p < 0.0024, ****p < 0.0001; tertiary: *p = 0.0321, ***p = 0.0006).

Article Snippet: We then incubated cells with goat-anti-rabbit-APC (Southern Biotech, Cat# 4050-11S; RRID: AB_2795959 ), mouse anti-human monoclonal antibodies against p75/NGFR (Becton-Dickinson, Cat# 40-1457) bound to phycoerythrin (PE), and EGFR (Fitzgerald, Acton, MA, Cat# 61R-E109BAF; RRID: AB_10808749 ) bound to FITC on ice in PBS/0.2% human serum for 30 min. After washing, we re-suspended cells in PBS/0.2% human serum containing 2 µg/mL 7-aminoactinomycin D (7-AAD) (Invitrogen, Cat# A1310).

Techniques: Microarray, Expressing, Derivative Assay, Western Blot, Immunohistochemistry, Staining, Fluorescence