Journal: bioRxiv

Article Title: Treatment with an Antigen-Specific Dual Microparticle System Reverses Advanced Multiple Sclerosis in Mice

doi: 10.1101/2022.03.25.485841

Figure Lengend Snippet: dMP MOG treatment in mice with advanced EAE results in reduction in MHCII components in microglia and CNS DCs, macrophages and B cells. Representative flow plots and quantification of CD74 in the spinal cord on day 7 in a) DCs; b) macrophages; c) microglia. d) MHC II in by B cells on Day 7. e) Multiplex immunofluorescence image of the lumbar spinal cord on day 17. Markers are as follows: CD4 (T helper cells) in green, CD8 (cytotoxic T cells) in yellow, CD74 (MHC II chaperone protein) in cyan, Ki67 (proliferation marker) in red, GFAP (astrocytes) in orange and Tmem119 (microglia) in magenta. Red arrows show APCs interacting with T Cells, white arrows point to astrocytes and gold arrows point to microglia. The dorsal column in the red box is magnified in the bottom image. All populations were pregated on live CD45+ DCs and Macrophages. B cells were negative for Tmem119 and CD3e. Macrophages and DCs were negative for B220/CD19, and macrophages were negative for CD11c. n=3-4/group * represents p<0.05 ** represents p <0.01

Article Snippet: Staining used the following antibodies, TMEM119 (Abcam, 28-3, HIER- EDTA pH 9.0, 1:100, dye 570), Ki67 (Abcam, SP6, HIER- Citrate pH 6.0, 1:300, dye 620), CD74 (LS Bio, Rb poly, HIER- EDTA pH 9.0, 1:600, dye 520), GFAP (Abcam, EPR1034Y, HIER- EDTA pH 9.0, 1:500, dye 540), CD4 (CST, D7D2Z, HIER- EDTA pH 9.0, 1:100, dye 650), and CD8 (CST, D4W2Z, HIER- EDTA pH 9.0, 1:100, dye 690).

Techniques: Multiplex Assay, Immunofluorescence, Marker

Journal: Journal of Neuropathology and Experimental Neurology

Article Title: Friedreich Ataxia: Developmental Failure of the Dorsal Root Entry Zone

doi: 10.1093/jnen/nlx087

Figure Lengend Snippet: Transverse and longitudinal sections of DR in FA. (A–D) Compound heterozygous FA (FA4, Table, GAA 896/c.11_12TCdel); (E, F) Homozygous FA (FA1, Table, GAA 1016/1016). (A) The transverse section shows multiple spherical eosinophilic expansions within the DR, measuring 20–50 µm in diameter. Some of the expansions contain nuclei. (B) The longitudinal section of this case shows a number of nuclei that are embedded in a fibrillary matrix. Most are consistent with Schwann cell nuclei, but one (arrow) lacks crenation and displays a clearer nucleoplasm, suggesting a non-Schwannian origin. (C–E) The expansions are strongly reactive with anti-GFAP (C), anti-S100 (D), and anti-vimentin (E), respectively. (F) An electron micrograph of an expansion reveals a heterogeneous collection of fibrillary bundles with variable main directions. The transverse diameter of the constituent filaments is 12.5–25 nm. Hematoxylin and eosin (A, B); GFAP (C); S100 (D); vimentin (E); electron micrograph (F). Scale bars: (A–E), 100 µm; (F), 200 nm.

Article Snippet: Sources of commercial antibodies; catalog numbers; host species and clonality; final protein concentrations or, in case of ascites fluid, dilution; and antigen retrieval methods were as follows: anti-PMP-22 (Abcam, Cambridge, MA, ab-31851, rabbit polyclonal, 2 μg protein/mL, incubation in 80% ethanol at 4 °C overnight); anti-P0 (Abcam, ab15508, rabbit polyclonal, 1 μg protein/mL, incubation in 80% ethanol at 4 °C overnight, followed by 0.01 M citric acid-sodium citrate buffer, pH 6, for 20 minutes at 95 °C); anti-PLP (Abcam, ab-105784, rabbit polyclonal, 2 μg protein/mL, incubation in 80% ethanol at 4 °C overnight, followed by 0.01 M citric acid-sodium citrate buffer, pH 6, for 20 minutes at 95 °C); anti-α-dystroglycan (Novus Biologicals, Littleton, CO, NBP1-49634, mouse monoclonal, 0.4 μg protein/mL, 0.01 M citric acid-sodium citrate buffer, pH 6, for 20 minutes at 95 °C); anti-periaxin (Novus, NBP1-89598, rabbit polyclonal, 2 μg protein/mL, 0.01 M citric acid-sodium citrate buffer, pH 6, 20 minutes at 95 °C); anti-laminin (Sigma-Aldrich, St. Louis, MO, L8271, mouse monoclonal, ascites fluid, diluted 1:1000, proteinase K [Sigma, P-6556], 0.1 μg/mL in tris buffer, pH 8, containing 0.01 M NaCl for 30 minutes at 37 °C); anti-GFAP (BioLegend, San Diego, CA, 837201, mouse monoclonal, ascites fluid, diluted 1:500, DIVA [a proprietary decloaking solution sold by Biocare Medical, Pacheco, CA] 1× for 30 minutes at 95 °C); anti-vimentin (Santa Cruz Biotechnology, Santa Cruz, CA, sc-66002, mouse monoclonal, 2 μg protein/mL, 0.01 M citric acid-sodium citrate buffer, pH 6, 20 minutes at 95 °C); anti-nestin (Santa Cruz, sc-23927, mouse monoclonal, 2 μg protein/mL, 0.01 M citric acid-sodium citrate buffer, pH 6, for 20 minutes at 95 °C, or DIVA for 30 minutes at 95 °C, or trypsin [DIFCO Laboratories, Detroit, MI, 0152-13] 1 mg/mL in 0.07 M CaCl 2 at pH 7.8 for 30 minutes at 37 °C); anti-S100 (Santa Cruz, sc-53438, mouse monoclonal, 0.8 μg protein/mL, 0.01 M citric acid-sodium citrate buffer, pH 6, 20 minutes at 95 °C); anti-class-III-β-tubulin (R&D Systems, Minneapolis, MN, MAB1195, mouse monoclonal, 2 μg protein/mL, 0.01 M citric acid-sodium citrate buffer, pH 6, 20 minutes at 95 °C); anti-phosphorylated neurofilament protein (Covance [now BioLegend] SMI-31 R, mouse monoclonal, ascites fluid, diluted 1:500, 0.01 M citric acid-sodium citrate buffer, pH 6, 20 minutes at 95 °C).

Techniques: