Journal: JCI Insight

Article Title: Role of the caspase-8/RIPK3 axis in Alzheimer’s disease pathogenesis and A β -induced NLRP3 inflammasome activation

doi: 10.1172/jci.insight.157433

Figure Lengend Snippet: ( A and B ) Representative images of the cortex, subiculum, and thalamus from 5-month-old WT and 5xFAD mice stained with Casp8 (orange) and Ripk3 (magenta) RNAscope probes. Scale bars: 40 μm. ( C ) Quantification of Casp8 staining. ( D ) Quantification of Ripk3 staining ( n = 5 5xFAD mice, n = 5 WT mice). ( E and F ) CASP8 and RIPK3 expression data obtained from a human transcriptomic data set (GSE33000) of dorsolateral prefrontal cortex tissue from 157 patients without dementia and 310 patients with AD. ( C – F ) Data were analyzed by Student’s t test. Data reported as mean ± SEM ( C and D ) or as violin plots ( E and F ). Ctrl, control.

Article Snippet: After amplification by sequential incubations with AMP1, 2, and 3 solutions (RNAscope Multiplex Fluorescent Detection Kit v2, Advanced Cell Diagnostics, catalog 323110) for 30 minutes at 40°C (separated with two 2-minute washes with wash buffer between solutions), slides were incubated for 15 minutes at 40°C with HRP-Channel 1 (RNAscope Multiplex Fluorescent Detection Kit v2, Advanced Cell Diagnostics, catalog 323110), before an additional 2 washes for 2 minutes in wash buffer.

Techniques: Staining, Expressing

Journal: Cell reports

Article Title: The adhesion GPCRs CELSR1–3 and LPHN3 engage G proteins via distinct activation mechanisms

doi: 10.1016/j.celrep.2023.112552

Figure Lengend Snippet:

Article Snippet: RNAscope Multiplex Fluorescent Assay: The following reagents were equilibrated at room temperature for 1 h: RNAscope Multiplex FL v2 AMP1 (Advanced Cell Diagnostics, 323101), RNAscope Multiplex FL v2 AMP2 (Advanced Cell Diagnostics, 323102), RNAscope Multiplex FL v2 AMP3 (Advanced Cell Diagnostics, 323103), RNAscope Multiplex FL v2 HRP C1 (Advanced Cell Diagnostics, 323104), RNAscope Multiplex FL v2 HRP C2 (Advanced Cell Diagnostics, 323105), RNAscope Multiplex FL v2 HRP C3 (Advanced Cell Diagnostics, 323106) and RNAscope Multiplex FL v2 HRP Blocker (Advanced Cell Diagnostics, 323107).

Techniques: Virus, Recombinant, Membrane, Electron Microscopy, Clone Assay, Software

Journal: Cell reports

Article Title: The adhesion GPCRs CELSR1–3 and LPHN3 engage G proteins via distinct activation mechanisms

doi: 10.1016/j.celrep.2023.112552

Figure Lengend Snippet:

Article Snippet: RNAscope Multiplex Fluorescent Assay: The following reagents were equilibrated at room temperature for 1 h: RNAscope Multiplex FL v2 AMP1 (Advanced Cell Diagnostics, 323101), RNAscope Multiplex FL v2 AMP2 (Advanced Cell Diagnostics, 323102), RNAscope Multiplex FL v2 AMP3 (Advanced Cell Diagnostics, 323103), RNAscope Multiplex FL v2 HRP C1 (Advanced Cell Diagnostics, 323104), RNAscope Multiplex FL v2 HRP C2 (Advanced Cell Diagnostics, 323105), RNAscope Multiplex FL v2 HRP C3 (Advanced Cell Diagnostics, 323106) and RNAscope Multiplex FL v2 HRP Blocker (Advanced Cell Diagnostics, 323107).

Techniques: Virus, Recombinant, Membrane, Electron Microscopy, Clone Assay, Software

Journal: Frontiers in Cellular Neuroscience

Article Title: Single-Cell Multiomic Approaches Reveal Diverse Labeling of the Nervous System by Common Cre-Drivers

doi: 10.3389/fncel.2021.648570

Figure Lengend Snippet: Wnt1-Cre2 and Sox10-Cre label neural crest and cells of the neural tube. (A) Wholemount and sequential transverse cross-sections of embryos in which ROSA26 Tomato/+ embryos were recombined using Sox10-Cre and (B) Wnt1-Cre. Axial positions of transverse cross-sections are noted for each Cre-driver. (C) Quantification of neural crest, (D) non-neural ectoderm and (E) neuroepithelial labeling obtained from Sox10-Cre and Wnt1-Cre.

Article Snippet: Mm-Wnt1-C2 RNAScope 2.5 LS Probe (ACD Bio 4011098-C2) was diluted 1:10 with Mm-Sox10 RNAScope 2.5 LS Probe (ACD Bio 435938) and the probe mixture was hybridized at 40°C for 2 h. Signal was amplified with RNAScope Multiplex FL v2 AMP1-3 for 30 min each at 40°C.

Techniques: Labeling

Journal: Frontiers in Cellular Neuroscience

Article Title: Single-Cell Multiomic Approaches Reveal Diverse Labeling of the Nervous System by Common Cre-Drivers

doi: 10.3389/fncel.2021.648570

Figure Lengend Snippet: Cells harvested using Wnt1-Cre2 have a neuroepithelial gene signature compared to cells harvested using Sox10-Cre. (A) Principal component analysis (PCA) plot and (B) heatmap normalized counts per million reads (CPM) showing that samples group together by Cre-driver. (C) Volcano plot of the significantly up- and downregulated genes obtained from comparing Wnt1-Cre2 to Sox10-Cre. (D) Gene ontology (GO) analysis of the significant differentially expressed genes. Categories were ordered by −log10( p -value) and dot size represents −log10( p -value). (E) Heatmap of normalized CPM values for neural crest and neuronal genes from cells harvested with Wnt1-Cre2 or Sox10-Cre.

Article Snippet: Mm-Wnt1-C2 RNAScope 2.5 LS Probe (ACD Bio 4011098-C2) was diluted 1:10 with Mm-Sox10 RNAScope 2.5 LS Probe (ACD Bio 435938) and the probe mixture was hybridized at 40°C for 2 h. Signal was amplified with RNAScope Multiplex FL v2 AMP1-3 for 30 min each at 40°C.

Techniques:

Journal: Frontiers in Cellular Neuroscience

Article Title: Single-Cell Multiomic Approaches Reveal Diverse Labeling of the Nervous System by Common Cre-Drivers

doi: 10.3389/fncel.2021.648570

Figure Lengend Snippet: Single-cell sequencing reveals little co-expression of Wnt1 and Sox10 in E9.5 neural crest cells. (A) UMAP plot showing expression of Wnt1 and Sox10 in cells of the E9.5 cranial region. (B) UMAP plot with Wnt1 - and Sox10 -expressing cells colored by cell population. (C) Bar plot of the percent of cells within the neural tube and neural crest expressing Wnt1 or Sox10 . (D) Blended UMAP plot showing little co-expression of Wnt1 and Sox10 . (E) Images of RNAScope for Sox10 and Wnt1 in E9.5 transverse cross-section through the cranial region showing co-localization in the delaminating neural crest, minimal co-localization in the migratory neural crest, and minimal expression of both Wnt1 and Sox10 in cells of the mesoderm. The location of the 63× image is noted within the 20× image. (F) Bar plot of differentially expressed genes and GO analysis of Wnt1 -positive cells and (G) Sox10 -positive cells. Categories were ordered by −log10( p -value) and dot size represents −log10( p -value).

Article Snippet: Mm-Wnt1-C2 RNAScope 2.5 LS Probe (ACD Bio 4011098-C2) was diluted 1:10 with Mm-Sox10 RNAScope 2.5 LS Probe (ACD Bio 435938) and the probe mixture was hybridized at 40°C for 2 h. Signal was amplified with RNAScope Multiplex FL v2 AMP1-3 for 30 min each at 40°C.

Techniques: Sequencing, Expressing

Journal: Frontiers in Cellular Neuroscience

Article Title: Single-Cell Multiomic Approaches Reveal Diverse Labeling of the Nervous System by Common Cre-Drivers

doi: 10.3389/fncel.2021.648570

Figure Lengend Snippet: Single-cell ATAC sequencing reveals accessible motifs near Wnt1 in the neuroepithelium and Sox10 in neural crest. (A) Coverage plot of accessibility at the Wnt1 locus in ectoderm-derived cell populations. (B) Bar plot of accessible motifs at the Wnt1 locus. (C) Dot plot showing expression of Otx2 and (D) Tead2 transcription factors with accessible predicted motifs at the Wnt1 locus. (E) Coverage plot of accessibility at the Sox10 locus in ectoderm-derived cell populations. (F) Bar plot of accessible motifs at the Sox10 locus. (G) Dot plot showing expression of Ets1 and (H) Ebf1 transcription factors with accessible predicted motifs at the Sox10 locus.

Article Snippet: Mm-Wnt1-C2 RNAScope 2.5 LS Probe (ACD Bio 4011098-C2) was diluted 1:10 with Mm-Sox10 RNAScope 2.5 LS Probe (ACD Bio 435938) and the probe mixture was hybridized at 40°C for 2 h. Signal was amplified with RNAScope Multiplex FL v2 AMP1-3 for 30 min each at 40°C.

Techniques: Sequencing, Derivative Assay, Expressing