rnascope 2.5 hd singleplex red chromogenic reagent kit  (Advanced Cell Diagnostics Inc)

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