Journal: Diagnostics

Article Title: Postmortem Cardiopulmonary Pathology in Patients with COVID-19 Infection: Single-Center Report of 12 Autopsies from Lausanne, Switzerland

doi: 10.3390/diagnostics11081357

Figure Lengend Snippet: Detection of actively transcribed SARS-CoV-2 in different cellular compartments of lung tissue. ( A ) In situ expression of SARS-CoV-2 mRNA (red), detected by RNAscope, in lung tissues from three infected patients with different levels of SARS-CoV-2 viral loads as detected by RT-qPCR in FFPE lung tissue. Granular positivity highlights hyaline membranes in RT-qPCR highly positive cases #3 and #6. Lack of in situ expression of SARS-CoV-2 mRNA in case #10, which showed also very low SARS-CoV-2 copy numbers in the RT-qPCR assay. ( B ) SARS-CoV-2 mRNA expression in pneumocytes and macrophages in two representative patients (cases #3 and #6), as detected by RNAscope and multiplex imaging on sequential lung tissue sections (magnifications as insets). Green arrows point to cells positive for viral mRNA. Cyan arrows highlight SARS-CoV-2 mRNA positive pneumocytes, defined by PANCK-expression. Magenta arrows depict SARS-CoV-2 mRNA positive macrophages, defined by CD68-positivity. ( C ) Lung tissue (case #6) stained for SARS-CoV2 mRNA expression (RNAscope). A simulated H&E image (generated from a fluorescent image processed using the inForm software) shows tissue areas positive for SARS-CoV-2 mRNA signal (red arrows) mainly associated with hyaline membranes. All images were acquired on the Vectra Polaris microscope using a 20× objective magnification.

Article Snippet: The RNAscope VS Universal HRP Reagent kit (Ref 323200, ACDBio) was used containing all required specific materials for standard target retrieval (CC1 24 min, recommended for lung tissue) and amplification of the targeted mRNA.

Techniques: In Situ, Expressing, RNAscope, Infection, Quantitative RT-PCR, Multiplex Assay, Imaging, Staining, Generated, Software, Microscopy

Journal: Cancer Immunology Research

Article Title: Host IL11 Signaling Suppresses CD4+ T cell–Mediated Antitumor Responses to Colon Cancer in Mice

doi: 10.1158/2326-6066.cir-19-1023

Figure Lengend Snippet: Figure 5. IL11RA expression negatively correlates with IFNG and CD4 coexpression in tumors of patients with colorectal cancer. Gene expression analysis was performed using three colon cancer datasets (GSE12945, GS17536, and GSE41258). A, Differential expression of IL11 and IL11RA between normal and tumor tissue. Student t test (, P < 0.0001). B and C, Representative images of RNAscope analysis of human colorectal cancer samples to spatially characterize IL11 and IL11RA expression. B, IL11 expression detected in NECs and CAFs. C, IL11RA expression in NECs, CAFs, and TILs. Scale bar, 100 mm. D, Representative images of IFNG and CD4 expression in IL11RA-high and -low tumors determined by RNAscope Fluorescent Multiplex analysis. CD4 (E) and IFNG (F) expression (pixel count relative to DAPI), and percentage of IFNG and CD4 colocalization (F) in tumors stratified by the extent of IL11RA expression as determined using the inForm software (PerkinElmer). Scale bar, 100 mm. Student t test (, P < 0.05). For all panels, data are expressed as mean SEM.

Article Snippet: RNA in situ hybridization—RNAscope TMAs constructed from histologically annotated primary human colorectal tumors, were sectioned at 4 mm onto SuperFrost Plus microscope slides and “baked” for 1 hour at 60 C. Singleplex RNAscope (catalog no. 322370; ACDBio) was performed on TMAs as per manufacturer’s instructions using the following paired oligonucleotide probes: Hs-IL11 (catalog no. 425289; ACDBio; NM_001267718.1; bp target region 131–1799 spanning exons 1–4) and Hs-IL11RA (catalog no. 568311; ACDBio; NM_001142784.2; bp target region 44–1177; spanning exons 1–11).

Techniques: Expressing, Gene Expression, Quantitative Proteomics, RNAscope, Multiplex Assay, Software