rnascope® protease plus reagent  (Advanced Cell Diagnostics Inc)

Journal: Nature

Article Title: Fibrin drives thromboinflammation and neuropathology in COVID-19

doi: 10.1038/s41586-024-07873-4

Figure Lengend Snippet: a , Representative RNAscope images of Trem2 , Cst7 , and Spp1 (red) mRNA expression in Iba-1 immunoreactive microglia (green) within the hippocampus of Delta-infected K18-hACE2 mice. b , Quantification of Trem2, Cst7, and Spp1 mRNA expression as percentage of total Iba-1+ cells in the hippocampus. Data are from n = 5 mice per group. Two-tailed Mann-Whitney test. Scale bars, 20 μm. All data are mean ± s.e.m. UI, uninfected.

Article Snippet: In brief, tissue was deparaffinized and incubated in 3% hydrogen peroxide for 10 min, then subjected to antigen retrieval by boiling in RNAScope Target Retrieval Solution (ACD Bio) for 1 h. The samples were permeabilized with RNAScope Protease Plus reagent (ACD Bio) for 30 min at 40 °C.

Techniques: RNAscope, Expressing, Infection, Two Tailed Test, MANN-WHITNEY

Journal: eLife

Article Title: Identification of bipotent progenitors that give rise to myogenic and connective tissues in mouse

doi: 10.7554/eLife.70235

Figure Lengend Snippet: ( A–D ) scRNAseq analysis of the Myf5 Cre/+ ; Rosa26 mTmG/+ E11.5 dataset (2 datasets of 2 embryos were aggregated to generate this data, see Materials and methods). ( A ) UMAPs of Myf5 Cre/+ ; Rosa26 mTmG/+ E11.5 RNA velocity trajectories. ( B ) Expression plots of Alx4 and Pax3 , highlighting EOM and Anterior somite clusters, respectively. ( C ) Plots of Myogenic and Non-myogenic signatures, Coexpression score and heatmaps of top markers, highlighting the transition population in EOM and anterior somites. Cells are ordered based on their non-myogenic signature score (increasing). The coexpression score is the product of the myogenic and non-myogenic signatures. Cells presenting a coexpression score higher than 0.20 are highlighted in yellow. These cells represent the transition between the myogenic and non-myogenic fates. ( D ) UMAP of the EOM subset revealing the bipartite fate of Myf5 -expressing cells. ( E-G’ ) RNAscope on Myf5 Cre/+ ; Rosa26 mTmG/+ E14.5 tissue sections with Pdgfra (cyan) and Pdgfa (red) probes ( E-E’’ ), Bmprb1 (red) and Bmp4 (cyan) probes ( F-F’ ) and Ephb1 (red) and Efnb1 (cyan) probes ( G-G’ ). Myf5 -derived cells are labelled by membrane GFP staining (n = 3 embryos each). Red and yellow arrowheads indicate Myf5 -derived myogenic and non-myogenic cells respectively. The dotted lines highlight the boundary of the muscle masses. ( H ) Quantification of the Ligand-Receptor scores for each pair (see Materials and methods). Note that these ratios are negative in the case of Bmp and Eph (signaling from non-myogenic to myogenic) but positive for Pdgf (signaling from myogenic to non-myogenic). ( G ) Model of myogenic and non-myogenic cell communication following bifurcation from a bipotent cell. Figure 4—source data 1. Excel table summarizing the quantification displayed on .

Article Snippet: Commercial assay, kit , RNAscope H202 & Protease Plus Reagents , ACD/Bio-techne , Cat #: 322330 , .

Techniques: Expressing, Derivative Assay, Membrane, Staining

Journal: eLife

Article Title: Identification of bipotent progenitors that give rise to myogenic and connective tissues in mouse

doi: 10.7554/eLife.70235

Figure Lengend Snippet: ( A ) UMAP of Myf5 Cre/+ ; Rosa26 mTmG/+ E11.5 EOM dataset illustrating velocity confidence and velocity length. Higher confidence is found on both ends of the EOM cluster. ( B ) Expression pattern of Scube1 in the EOM subset (mostly in the non-myogenic compartment). ( C-C’’ ) Combined RNAscope for Scube1 and Myod/Myog/GFP immunostaining on transverse sections of Myf5 Cre/+ ; Rosa26 mTmG/+ at E14.5. ( D ) Model of the EOM muscle vs EOM origin compartmentalization used for quantification in (E). ( E ) Quantification of Scube1 signal in each compartment (n = 2 embryos). Figure 5—figure supplement 1—source data 1. Excel table summarizing the quantification displayed on .

Article Snippet: Commercial assay, kit , RNAscope H202 & Protease Plus Reagents , ACD/Bio-techne , Cat #: 322330 , .

Techniques: Expressing, Immunostaining