Journal: Cell reports

Article Title: In Situ Fixation Redefines Quiescence and Early Activation of Skeletal Muscle Stem Cells.

doi: 10.1016/j.celrep.2017.10.080

Figure Lengend Snippet: Figure 1. Isolation of MuSCs in a Fixed, G0-Arrested State (A) Graphical scheme of the in situ fixation protocol for MuSCs isolation and comparison to the standard protocol. A detailed description of the protocols is available in the Supplemental Information. BM, basement membrane; T0-SC, time-zero/quiescent MuSC; T3-SC, time 3 hr/activated MuSC. (B) FACS profiles of non-fixed (T3-SC) and fixed (T0-SC) GFP+ MuSCs from Tg:Pax7-nGFP muscle preparations. Sorted GFP+ cells are marked as red dots in all plots. SSC, side scatter, FSC, forward scatter. Values on the plots indicate mean percentage of sorted GFP+ cells of the total number of events, excluding small SSC/FSC and doublets; n = 5. (C) Proliferation of FACS-isolated T3-SCs and T0-SCs cultured for 48 hr and stained with EdU (24-hr chase); n = 3. EdU+ cells: 0% for T0-SCs and 74% ± 0.03% for T3-SCs. (D) Nascent RNA synthesis in FACS-isolated T3-SCs and T0-SCs cultured for 48 hr and incubated with labeled ethynyl uridine (EU) ribonucleoside for 2 hr; n = 3. Average of 80 nuclei counted per sample, 100% EU+ and EU for T3-SC and T0-SC, respectively. Dotted line delineates cell’s nucleus. (E) Morphology of T3 and T0 MuSCs immediately after the FACS. Cells were spun on Matrigel-coated slides, and membranes were stained with the lectin marker WGA (wheat germ agglutinin). Mean eccentricity of the cells was computationally calculated (Experimental Procedures). Data are reported as mean ± SD; n.d, not detected. Scale bars, 90 mm in (C), 20 mm in (D), 8 mm in (D, insets), and 10 mm in (E). See also Supplemental Information.

Article Snippet: The purity of the FACS-isolated cells was assessed with anti-PAX7 antibody (mouse monoclonal; Santa Cruz Biotechnology, #sc-81648) diluted 1:100 in BS and incubated for 16 hr at 37 C. For the membrane staining, WGA was used following manufacturer’s guidelines (Thermo Fisher Scientific, #W32464) on T0- or T3-SCs after a 15-min centrifugation (300 3 g, 4 C) on Matrigel (Corning, #354248)-coated 8-chamber slides (Sarstedt, #94.6140.802).

Techniques: Isolation, In Situ, Comparison, Membrane, Cell Culture, Staining, Incubation, Labeling, Marker

Journal: Cell reports

Article Title: In Situ Fixation Redefines Quiescence and Early Activation of Skeletal Muscle Stem Cells.

doi: 10.1016/j.celrep.2017.10.080

Figure Lengend Snippet: Figure 5. Histone H3 Modifications and DNA Methylation during Early MuSCs Activation (A) Left: H3K4me3 ChIP-seq averaged signal at promoters. Right: overlap between T0-SC and T3-SC genes with H3K4me3 peaks at promoters. TSS, tran- scription start site. (B) Left: H3K27ac ChIP-seq averaged signal at active distal enhancers (H3K27ac peaks outside gene body) from T0-SC and T3-SC samples. Right: overlap of genes that present at least one distal enhancer between T0-SCs and T3-SCs. (C) Examples of H3K4me3 and H3K27ac ChIP-seq tracks from T3 and T0 upregulated genes Fosl and Egr3. (D) Examples of H3K4me3 and H3K27ac ChIP-seq tracks from T3 and T0 downregulated genes Hes1 and Pax7. (E) H3K27me3 ChIP-seq averaged signal at promoters between T0-SCs, T3-SCs, and activated MuSCs from a published dataset (Liu et al., 2013). (F) Volcano plot representing genomic DNA methylation changes between T0-SCs and T3-SCs. Each dot represents one of 27,878 CpG clusters interrogated. Among all clusters (blue), those with a methylation change R 10% and an FDR < 0.01 were considered significantly different (red). FDR, false discovery rate (logarithmic scale).

Article Snippet: The purity of the FACS-isolated cells was assessed with anti-PAX7 antibody (mouse monoclonal; Santa Cruz Biotechnology, #sc-81648) diluted 1:100 in BS and incubated for 16 hr at 37 C. For the membrane staining, WGA was used following manufacturer’s guidelines (Thermo Fisher Scientific, #W32464) on T0- or T3-SCs after a 15-min centrifugation (300 3 g, 4 C) on Matrigel (Corning, #354248)-coated 8-chamber slides (Sarstedt, #94.6140.802).

Techniques: DNA Methylation Assay, Activation Assay, ChIP-sequencing, Methylation

Journal: PLoS Neglected Tropical Diseases

Article Title: Diagnostic accuracy of serological tests for the diagnosis of Chikungunya virus infection: A systematic review and meta-analysis

doi: 10.1371/journal.pntd.0010152

Figure Lengend Snippet: Characteristics of studies on IgM detection tests included in the meta-analysis.

Article Snippet: On-site CHIK IgM Combo Rapid test , CTK Biotech Inc., San Diego, CA, USA , 3 , 145 , 27.9 [10.8; 55.2] , 81.0% [40.5; 93.9] , 90.4 , 98.7 [84.9; 99.9] , 0.0% [0.0; 0.0] , 98.

Techniques: Enzyme-linked Immunosorbent Assay, Indirect ELISA, HI Assay, Plaque Reduction Neutralization Test, Neutralization

Journal: PLoS Neglected Tropical Diseases

Article Title: Diagnostic accuracy of serological tests for the diagnosis of Chikungunya virus infection: A systematic review and meta-analysis

doi: 10.1371/journal.pntd.0010152

Figure Lengend Snippet: Subgroup analysis for commercial tests.

Article Snippet: On-site CHIK IgM Combo Rapid test , CTK Biotech Inc., San Diego, CA, USA , 3 , 145 , 27.9 [10.8; 55.2] , 81.0% [40.5; 93.9] , 90.4 , 98.7 [84.9; 99.9] , 0.0% [0.0; 0.0] , 98.

Techniques: Enzyme-linked Immunosorbent Assay, Virus, Immunofluorescence