Journal: iScience

Article Title: Circadian disruption during fetal development promotes pathological cardiac remodeling in male mice

doi: 10.1016/j.isci.2024.109008

Figure Lengend Snippet:

Article Snippet: Rat monoclonal Anti-Mouse UGRP1/SCGB3A2 (IgG2B Clone # 381707) , R and D Systems , Cat# MAB3465.

Techniques: Recombinant, Virus, Fluorescence, Bicinchoninic Acid Protein Assay, cDNA Synthesis, SYBR Green Assay, Enzyme-linked Immunosorbent Assay, Software

Journal: The European Respiratory Journal

Article Title: Airway-derived emphysema-specific alveolar type II cells exhibit impaired regenerative potential in COPD

doi: 10.1183/13993003.02071-2023

Figure Lengend Snippet: Alveolar epithelial cell heterogeneity in COPD. a) Sample information of single-cell RNA sequencing (scRNAseq) data. b) Schematic showing workflow of sampling parenchymal tissue from emphysematous (n=6) and healthy (n=4) lungs, viable epithelial cell isolation and scRNAseq using the 10X Genomics platform. Uniform manifold approximation and projection (UMAP) shows cells obtained from COPD patients and healthy donors. c) UMAP visualisation of annotated lung epithelial cell types. d) The cellular composition of epithelial cells from individual COPD patients and healthy donors. e) The numbers of genes upregulated (log 2 fold change (fc) >1, p<0.05) and downregulated (log 2 fc <−1, p<0.05) in each cell type from COPD patients, compared to those of healthy donor. f) UMAP visualisation of alveolar type II (ATII) cells marked by expression of surfactant protein C (SFTPC). g) UMAP showing ATII cells from emphysema patients and healthy donors. h) Feature plots showing expression of secretoglobins SCGB3A2 , SCGB3A1 and SCGB1A1 in ATII cells. NA: not applicable; GOLD: Global Initiative for Chronic Obstructive Lung Disease; MACS: magnetic-activated cell sorting; FACS: fluorescence-activated cell sorting; EpCAM: epithelial cell adhesion molecule; NEC: nonepithelial cells; DEGs: differentially expressed genes.

Article Snippet: Primary antibodies used for immunofluorescence include rabbit anti-SPC (Millipore, AB3786), rabbit anti-SPC (Abcam, ab90716), mouse IgM anti-HTII280 (Terrace Biotech, TB-27AHT2-280), CD74 (Novus, NBP1-33109), intercellular adhesion molecule 1 (ICAM1) (Novus, BBA3), TM4SF1 (R&D Systems, MAB8164), SCGB3A2 (R&D Systems, AF3545), acetylated α-tubulin (Abcam, ab24610), tdTomato (Biorbyt, orb182397) and chicken anti-Ly6i antibody.

Techniques: RNA Sequencing, Sampling, Cell Isolation, Expressing, FACS, Fluorescence

Journal: The European Respiratory Journal

Article Title: Airway-derived emphysema-specific alveolar type II cells exhibit impaired regenerative potential in COPD

doi: 10.1183/13993003.02071-2023

Figure Lengend Snippet: Identification of secretaglobin (SCGB pos ) alveolar type II (ATII) cell subpopulations in human COPD lungs. a) Haematoxylin and eosin (H&E) staining of emphysematous parenchymal lung tissue from COPD patient at low magnification. Scale bar=200 µm. Within thin alveolar sacs (inset), fluorescent RNAScope for SCGB3A2 (red), 1A1 (green), 3A1 (green) and immunofluorescence (IF) staining for surfactant protein C (SPC) (white) identifies 3A2 + ATII cells (arrowheads). Scale bars=40 µm. b) H&E staining of respiratory bronchiole region (inset) in COPD lung tissue. Scale bar=100 µm. Multiplex fluorescence RNAScope for SCGB3A2 (red), 1A1 (green), 3A1 (green), and co-IF for SPC (white) identifies ATII cells co-expressing multiple SCGB genes (arrowheads) in this region. Scale bars=40 µm. c) Quantification of percentages of 3A2 + and 1A1 + /3A1 + /3A2 + ATII cells detected by RNAScope and IF in alveolar sacs of healthy (n=5) and emphysematous (n=9) lung tissue. d) Uniform manifold approximation and projection (UMAP) showing annotation of ATII subclusters based on expressions of SCGB genes and tissue localisations. ATII cells dominantly presenting in alveolar sacs of COPD samples and expressing SCGB3A2 were identified as asATII. ATII cells expressing multiple SCGBs and localised in respiratory bronchioles were annotated as rbATII. The remaining of ATII cells, dominantly presented in healthy samples, were marked as hATII. e) Percentages of ATII subclusters in healthy and COPD samples. f) Dot plot of selected senescence marker genes in asATII, rbATII and hATII cells. g) Dot plot of selected marker genes identified in asATII and rbATII cells. ns : nonsignificant. ***: p<0.001. ns : p>0.05, Mann–Whitney test.

Article Snippet: Primary antibodies used for immunofluorescence include rabbit anti-SPC (Millipore, AB3786), rabbit anti-SPC (Abcam, ab90716), mouse IgM anti-HTII280 (Terrace Biotech, TB-27AHT2-280), CD74 (Novus, NBP1-33109), intercellular adhesion molecule 1 (ICAM1) (Novus, BBA3), TM4SF1 (R&D Systems, MAB8164), SCGB3A2 (R&D Systems, AF3545), acetylated α-tubulin (Abcam, ab24610), tdTomato (Biorbyt, orb182397) and chicken anti-Ly6i antibody.

Techniques: Staining, RNAscope, Immunofluorescence, Multiplex Assay, Fluorescence, Expressing, Marker, MANN-WHITNEY

Journal: The European Respiratory Journal

Article Title: Airway-derived emphysema-specific alveolar type II cells exhibit impaired regenerative potential in COPD

doi: 10.1183/13993003.02071-2023

Figure Lengend Snippet: Decreased progenitor cell function of emphysema-specific alveolar type II (ATII) cells in human COPD lungs. a) Detection of asATII cells in alveolar sacs of emphysematous region (black square in left picture with haematoxylin and eosin (H&E) staining) by co-immunofluorescence (co-IF) for CD74 (green), secretaglobin-3A2 (SCGB3A2) (red), and surfactant protein C (SPC) (white). Scale bars=200 µm, 50 µm and 10 µm. b) Detection of rbATII cells in respiratory bronchiole of emphysematous region (black square in left picture with H&E staining) by co-IF for intercellular adhesion molecule 1 (ICAM1) (green, top), TM4SF1 (green, bottom), SCGB3A2 (red), and SPC (white). Scale bars=50 µm, 20 µm and 10 µm. c) Flow cytometry analysis of asATII and rbATII using CD74 (centre), TM4SF1 and ICAM1 (right) in healthy control sample (n=6, top) and emphysematous parenchymal tissue from COPD patients (n=6, bottom). d) Quantification of the percentages of asATII (CD74 + ) and rbATII (ICAM1 + /TM4SF1 + ) cells in total ATII cells (HTII280 + /epithelial cell adhesion molecule (EPCAM) + /4′,6-diamidino-2-phenylindole (DAPI) − ) in a). Mann–Whitney test. e) Organoids formed by fluorescence-activated cell sorted asATII (HTII280 + /CD74 + ) and rbATII (HTII280 + /ICAM1 + /TM4SF1) cells from human emphysematous parenchymal tissue (n=5) and by ATII cells (HTII280 + ) from healthy control (n=3). Scale bar=100 µm. f) Quantification of organoid forming efficiency of c). Values are presented as mean± sem . Mann–Whitney test. *: p<0.05.

Article Snippet: Primary antibodies used for immunofluorescence include rabbit anti-SPC (Millipore, AB3786), rabbit anti-SPC (Abcam, ab90716), mouse IgM anti-HTII280 (Terrace Biotech, TB-27AHT2-280), CD74 (Novus, NBP1-33109), intercellular adhesion molecule 1 (ICAM1) (Novus, BBA3), TM4SF1 (R&D Systems, MAB8164), SCGB3A2 (R&D Systems, AF3545), acetylated α-tubulin (Abcam, ab24610), tdTomato (Biorbyt, orb182397) and chicken anti-Ly6i antibody.

Techniques: Cell Function Assay, Staining, Immunofluorescence, Flow Cytometry, Control, MANN-WHITNEY, Fluorescence