in situ hybridization rnascope multiplex fluorescent kit  (Advanced Cell Diagnostics Inc)

Journal: Journal of respiratory biology and translational medicine

Article Title: Enhanced Spatial Transcriptomics Analysis of Mouse Lung Tissues Reveals Cell-Specific Gene Expression Changes Associated with Pulmonary Hypertension

doi: 10.70322/jrbtm.2025.10004

Figure Lengend Snippet: Spatial plot showing the change of EC subpopulation in PH mice. ( A ) Spatial plots showing the arterial ECs (AEC) and endothelial progenitor cells (EPC) were increased in KO mice ( n = 2). ( B ) Quantification of AECs and EPCs cell proportions in ( A ). ( C ) Spatial plots showing the general capillary ECs (CAP1) and aerocytes (CAP2) were reduced in KO mice ( n = 2). ( D ) Quantification data of AECs and EPCs in ( C ). ( E ) Spatial and Violin plots showing the increased AEC marker Sox17 , decreased CAP1 (gCap) marker Plvap , and decreased CAP2 (aCap) marker Prx in the KO mice ( n = 2). *** p < 0.001, **** p < 0.0001. ( F ) Immunostaining and RNASCOPE analysis validated the reduction of CAP2 ( Ednrb + /CD31 + ) in KO mice (WT n = 4, KO n = 3). * p < 0.05.

Article Snippet: A Multiplex Fluorescent V2 RNAscope in situ hybridization (Advanced Cell Diagnostics, Newark, CA, USA) and immunostaining assay were conducted on lung cryosections obtained from mouse lung samples.

Techniques: Marker, Immunostaining, RNAscope

Journal: Nature Communications

Article Title: Conserved transcriptional regulation by BRN1 and BRN2 in neocortical progenitors drives mammalian neural specification and neocortical expansion

doi: 10.1038/s41467-024-52443-x

Figure Lengend Snippet: a UMAPs from control (CT) and Brn1/2-cKO (cKO) cortices at E12.5 and E14.5 by TRIcycle (Transferable Representation and Inference of cell cycle) analysis . b Cell cycle phase analysis from control and Brn1/2-cKO apical progenitors (AP) and basal progenitors (BP) at E12.5 and E14.5 represented as cell density (Wilcoxon rank sum test: AP-E12.5 p = 0.085; AP-E14.5 p = 1.84e−15; BP-E12.5 p = 1.95e-06; BP-E14.5 p < 2.2e−16). Empty arrowheads point to reduced S-G2/M state in mutants, arrowheads to increased G1/G0 state. c, f Schematic of the experimental strategy. E12.5 and E14.5 cortices were analyzed by EdU and Ki67 immunolabeling 1 h ( c ) or 24 h ( f ) after intraperitoneal injection of EdU. d EdU (red) and Ki67 (grey) immunolabeling in control and Brn1/2-cKO after 1 h EdU injection at E12.5 and E14.5 (E12.5: n = 5 CT, n = 3 cKO mice; E14.5: n = 5 CT, n = 4 cKO mice; two-sided unpaired t- test: E12.5 Ki67 p = 0.1212, E12.5 EdU p = 0.0253, E14.5 Ki67 p = 0.0008, E14.5 EdU p = 0.0053). e EdU labeling index in control and Brn1/2-cKO after 1 h EdU injection at E12.5 and E14.5 (E12.5: n = 4 CT, n = 3 cKO mice; E14.5: n = 5 CT, n = 4 cKO mice; two-sided unpaired t- test: E12.5 p = 0.5296, E14.5 p = 0.4063). g EdU (red) and Ki67 (grey) immunolabeling in control and Brn1/2-cKO after 24 h EdU injection at E12.5 and E14.5 (E12.5: n = 3 CT, n = 5 cKO mice; E14.5: n = 4 mice/group; two-sided unpaired t- test: E12.5 p = 0.0135, E14.5 p = 0.0071). Boxed area at higher magnification on the right. Lines and dashed lines circulating the cells show expression or absence of Ki67, respectively. h Schematic of progenitors dividing and differentiating into neurons. i ScRNAseq expression of Ezh2 and Ngn2 at E14.5 in control and Brn1/2-cKO APs and BPs. j RNAscope for Ezh2 (red) and Ngn2 (grey) in control and Brn1/2-cKO cortices at E14.5 ( n = 3 mice/group; two-sided unpaired t- test: Ezh2 -p = 0.0357, Ngn2 -p = 0.0436). Low and top lines represent the limits of the ventricular zone (VZ) and cortical plate (CP), respectively. SVZ Subventricular Zone, IP Intermediate Progenitors, N Neurons. Values are mean ± SEM; * p < 0.05, ** p < 0.01, *** p < 0.001; Scale bars: 50 µm (lower magnification), 10 µm (higher magnification). Source data are provided as a Source Data file.

Article Snippet: Brain sections were processed for multiplex fluorescent in situ hybridization RNAscope following the manufacturer’s instructions (#323110, Advanced Cell Diagnostics).

Techniques: Control, Cell Cycle Assay, Immunolabeling, Injection, Labeling, Expressing, RNAscope