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


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    Advanced Cell Diagnostics Inc rnascope 2.5 hd singleplex reagent kit
    Rnascope 2.5 Hd Singleplex Reagent Kit, supplied by Advanced Cell Diagnostics Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/rnascope+2%2E5+hd+singleplex+reagent+kit/rnascope+multiplex+fluorescent+reagent+kit+v2/pmc10228551-115-7-13
    Average 90 stars, based on 1 article reviews
    rnascope 2.5 hd singleplex reagent kit - by Bioz Stars, 2026-11
    90/100 stars

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    Related Articles

    RNAscope:

    Article Title: Dorsal root ganglion-targeted analgesic delivery for effective relief of neuropathic pain
    Article Snippet: RNA-scope assay was performed for in situ evaluation of Adam8 (Advanced Cell Diagnostics, #1047421-C1) in the DRG of the mice using RNA-Scope 2.5 HD Red Kit (Advanced Cell Diagnostics, #322360) following our recent work [ , ].

    Article Title: Prdm16 regulates the postnatal fate of embryonic radial glia via Vcam1-dependent mechanisms
    Article Snippet: We then washed the whole-mounts 3 times with PBS with 0.1% Triton-X100, blocked in 10% normal donkey serum in PBS with 2% Triton-X100 at room temperature for 1 h, and stained with primary antibodies against Vcam1 (BD Biosciences, 550547, 1:100), γ-Tubulin (Sigma, T5192-25UL, 1:1000), and SOX2 (R&D Systems, AF2018, 1:1000) for 48 h at 4 °C.

    Article Title: Nr4a1 modulates inflammation and heart regeneration in zebrafish
    Article Snippet: The probe synthesis was conducted using the Invitrogen MEGAscript T7 Transcription Kit with the DIG RNA labeling mix (Roche).

    Article Title: mRNA-based in vivo induction of ETV2 to restore blood flow in a preclinical mouse hindlimb ischemia model
    Article Snippet: In situ hybridization (ISH) was performed using an RNAscope 2.5 LS Reagent Kit-BROWN (Advanced Cell Diagnostics [ACD], Hayward, CA, cat no. 322100) for use with BOND RX Stainer (Leica Biosystems, Buffalo Grove, IL) according to the manufacturer’s instructions.

    Article Title: Notum as a Crucial Regulator of Matrix Integrity in Dentinogenesis
    Article Snippet: In situ hybridization was performed with the RNAscope 2.5 High‐Definition Assay‐Brown (Advanced Cell Diagnostics, Newark, CA, USA) according to the user manual 322452 (FFPE sample preparation and pretreatment) and 322310 (RNAscope 2.5 High‐Definition Detection Reagent‐Brown user manual) provided by the manufacturer.

    Article Title: Osteoclast-independent osteocyte dendrite defects in mice bearing the osteogenesis imperfecta-causing Sp7 R342C mutation
    Article Snippet: Sections were then incubated with phalloidin mixture (1:1 000, Abcam) in the dark for 1 h at room temperature.

    Article Title: Maladaptive inflammatory signaling in old mice impairs colonic regeneration by promoting a sustained fetal-like epithelial state
    Article Snippet: RNA in situ hybridization for Lgr5 expression was conducted on 7 μm paraffin sections using the RNAscope 2.5 High Definition (HD) – Red Assay (Advanced Cell Diagnostics, 322350).

    In Situ Hybridization:

    Article Title: Dorsal root ganglion-targeted analgesic delivery for effective relief of neuropathic pain
    Article Snippet: RNA-scope assay was performed for in situ evaluation of Adam8 (Advanced Cell Diagnostics, #1047421-C1) in the DRG of the mice using RNA-Scope 2.5 HD Red Kit (Advanced Cell Diagnostics, #322360) following our recent work [ , ].

    Article Title: Prdm16 regulates the postnatal fate of embryonic radial glia via Vcam1-dependent mechanisms
    Article Snippet: We then washed the whole-mounts 3 times with PBS with 0.1% Triton-X100, blocked in 10% normal donkey serum in PBS with 2% Triton-X100 at room temperature for 1 h, and stained with primary antibodies against Vcam1 (BD Biosciences, 550547, 1:100), γ-Tubulin (Sigma, T5192-25UL, 1:1000), and SOX2 (R&D Systems, AF2018, 1:1000) for 48 h at 4 °C.

    Article Title: Nr4a1 modulates inflammation and heart regeneration in zebrafish
    Article Snippet: The probe synthesis was conducted using the Invitrogen MEGAscript T7 Transcription Kit with the DIG RNA labeling mix (Roche).

    Article Title: mRNA-based in vivo induction of ETV2 to restore blood flow in a preclinical mouse hindlimb ischemia model
    Article Snippet: In situ hybridization (ISH) was performed using an RNAscope 2.5 LS Reagent Kit-BROWN (Advanced Cell Diagnostics [ACD], Hayward, CA, cat no. 322100) for use with BOND RX Stainer (Leica Biosystems, Buffalo Grove, IL) according to the manufacturer’s instructions.

    Article Title: Notum as a Crucial Regulator of Matrix Integrity in Dentinogenesis
    Article Snippet: In situ hybridization was performed with the RNAscope 2.5 High‐Definition Assay‐Brown (Advanced Cell Diagnostics, Newark, CA, USA) according to the user manual 322452 (FFPE sample preparation and pretreatment) and 322310 (RNAscope 2.5 High‐Definition Detection Reagent‐Brown user manual) provided by the manufacturer.

    Article Title: Osteoclast-independent osteocyte dendrite defects in mice bearing the osteogenesis imperfecta-causing Sp7 R342C mutation
    Article Snippet: Sections were then incubated with phalloidin mixture (1:1 000, Abcam) in the dark for 1 h at room temperature.

    Article Title: Maladaptive inflammatory signaling in old mice impairs colonic regeneration by promoting a sustained fetal-like epithelial state
    Article Snippet: RNA in situ hybridization for Lgr5 expression was conducted on 7 μm paraffin sections using the RNAscope 2.5 High Definition (HD) – Red Assay (Advanced Cell Diagnostics, 322350).

    Multiplex Assay:

    Article Title: Dorsal root ganglion-targeted analgesic delivery for effective relief of neuropathic pain
    Article Snippet: RNA-scope assay was performed for in situ evaluation of Adam8 (Advanced Cell Diagnostics, #1047421-C1) in the DRG of the mice using RNA-Scope 2.5 HD Red Kit (Advanced Cell Diagnostics, #322360) following our recent work [ , ].

    Article Title: Prdm16 regulates the postnatal fate of embryonic radial glia via Vcam1-dependent mechanisms
    Article Snippet: We then washed the whole-mounts 3 times with PBS with 0.1% Triton-X100, blocked in 10% normal donkey serum in PBS with 2% Triton-X100 at room temperature for 1 h, and stained with primary antibodies against Vcam1 (BD Biosciences, 550547, 1:100), γ-Tubulin (Sigma, T5192-25UL, 1:1000), and SOX2 (R&D Systems, AF2018, 1:1000) for 48 h at 4 °C.

    Article Title: Nr4a1 modulates inflammation and heart regeneration in zebrafish
    Article Snippet: The probe synthesis was conducted using the Invitrogen MEGAscript T7 Transcription Kit with the DIG RNA labeling mix (Roche).

    Article Title: mRNA-based in vivo induction of ETV2 to restore blood flow in a preclinical mouse hindlimb ischemia model
    Article Snippet: In situ hybridization (ISH) was performed using an RNAscope 2.5 LS Reagent Kit-BROWN (Advanced Cell Diagnostics [ACD], Hayward, CA, cat no. 322100) for use with BOND RX Stainer (Leica Biosystems, Buffalo Grove, IL) according to the manufacturer’s instructions.

    Article Title: Notum as a Crucial Regulator of Matrix Integrity in Dentinogenesis
    Article Snippet: In situ hybridization was performed with the RNAscope 2.5 High‐Definition Assay‐Brown (Advanced Cell Diagnostics, Newark, CA, USA) according to the user manual 322452 (FFPE sample preparation and pretreatment) and 322310 (RNAscope 2.5 High‐Definition Detection Reagent‐Brown user manual) provided by the manufacturer.

    Article Title: Osteoclast-independent osteocyte dendrite defects in mice bearing the osteogenesis imperfecta-causing Sp7 R342C mutation
    Article Snippet: Sections were then incubated with phalloidin mixture (1:1 000, Abcam) in the dark for 1 h at room temperature.

    Article Title: Maladaptive inflammatory signaling in old mice impairs colonic regeneration by promoting a sustained fetal-like epithelial state
    Article Snippet: RNA in situ hybridization for Lgr5 expression was conducted on 7 μm paraffin sections using the RNAscope 2.5 High Definition (HD) – Red Assay (Advanced Cell Diagnostics, 322350).

    In Situ:

    Article Title: Dorsal root ganglion-targeted analgesic delivery for effective relief of neuropathic pain
    Article Snippet: RNA-scope assay was performed for in situ evaluation of Adam8 (Advanced Cell Diagnostics, #1047421-C1) in the DRG of the mice using RNA-Scope 2.5 HD Red Kit (Advanced Cell Diagnostics, #322360) following our recent work [ , ].

    Article Title: Prdm16 regulates the postnatal fate of embryonic radial glia via Vcam1-dependent mechanisms
    Article Snippet: We then washed the whole-mounts 3 times with PBS with 0.1% Triton-X100, blocked in 10% normal donkey serum in PBS with 2% Triton-X100 at room temperature for 1 h, and stained with primary antibodies against Vcam1 (BD Biosciences, 550547, 1:100), γ-Tubulin (Sigma, T5192-25UL, 1:1000), and SOX2 (R&D Systems, AF2018, 1:1000) for 48 h at 4 °C.

    Article Title: Nr4a1 modulates inflammation and heart regeneration in zebrafish
    Article Snippet: The probe synthesis was conducted using the Invitrogen MEGAscript T7 Transcription Kit with the DIG RNA labeling mix (Roche).

    Article Title: mRNA-based in vivo induction of ETV2 to restore blood flow in a preclinical mouse hindlimb ischemia model
    Article Snippet: In situ hybridization (ISH) was performed using an RNAscope 2.5 LS Reagent Kit-BROWN (Advanced Cell Diagnostics [ACD], Hayward, CA, cat no. 322100) for use with BOND RX Stainer (Leica Biosystems, Buffalo Grove, IL) according to the manufacturer’s instructions.

    Article Title: Notum as a Crucial Regulator of Matrix Integrity in Dentinogenesis
    Article Snippet: In situ hybridization was performed with the RNAscope 2.5 High‐Definition Assay‐Brown (Advanced Cell Diagnostics, Newark, CA, USA) according to the user manual 322452 (FFPE sample preparation and pretreatment) and 322310 (RNAscope 2.5 High‐Definition Detection Reagent‐Brown user manual) provided by the manufacturer.

    Article Title: Osteoclast-independent osteocyte dendrite defects in mice bearing the osteogenesis imperfecta-causing Sp7 R342C mutation
    Article Snippet: Sections were then incubated with phalloidin mixture (1:1 000, Abcam) in the dark for 1 h at room temperature.

    Article Title: Maladaptive inflammatory signaling in old mice impairs colonic regeneration by promoting a sustained fetal-like epithelial state
    Article Snippet: RNA in situ hybridization for Lgr5 expression was conducted on 7 μm paraffin sections using the RNAscope 2.5 High Definition (HD) – Red Assay (Advanced Cell Diagnostics, 322350).

    Formalin-fixed Paraffin-Embedded:

    Article Title: Dorsal root ganglion-targeted analgesic delivery for effective relief of neuropathic pain
    Article Snippet: RNA-scope assay was performed for in situ evaluation of Adam8 (Advanced Cell Diagnostics, #1047421-C1) in the DRG of the mice using RNA-Scope 2.5 HD Red Kit (Advanced Cell Diagnostics, #322360) following our recent work [ , ].

    Article Title: Prdm16 regulates the postnatal fate of embryonic radial glia via Vcam1-dependent mechanisms
    Article Snippet: We then washed the whole-mounts 3 times with PBS with 0.1% Triton-X100, blocked in 10% normal donkey serum in PBS with 2% Triton-X100 at room temperature for 1 h, and stained with primary antibodies against Vcam1 (BD Biosciences, 550547, 1:100), γ-Tubulin (Sigma, T5192-25UL, 1:1000), and SOX2 (R&D Systems, AF2018, 1:1000) for 48 h at 4 °C.

    Article Title: Nr4a1 modulates inflammation and heart regeneration in zebrafish
    Article Snippet: The probe synthesis was conducted using the Invitrogen MEGAscript T7 Transcription Kit with the DIG RNA labeling mix (Roche).

    Article Title: mRNA-based in vivo induction of ETV2 to restore blood flow in a preclinical mouse hindlimb ischemia model
    Article Snippet: In situ hybridization (ISH) was performed using an RNAscope 2.5 LS Reagent Kit-BROWN (Advanced Cell Diagnostics [ACD], Hayward, CA, cat no. 322100) for use with BOND RX Stainer (Leica Biosystems, Buffalo Grove, IL) according to the manufacturer’s instructions.

    Article Title: Notum as a Crucial Regulator of Matrix Integrity in Dentinogenesis
    Article Snippet: In situ hybridization was performed with the RNAscope 2.5 High‐Definition Assay‐Brown (Advanced Cell Diagnostics, Newark, CA, USA) according to the user manual 322452 (FFPE sample preparation and pretreatment) and 322310 (RNAscope 2.5 High‐Definition Detection Reagent‐Brown user manual) provided by the manufacturer.

    Article Title: Osteoclast-independent osteocyte dendrite defects in mice bearing the osteogenesis imperfecta-causing Sp7 R342C mutation
    Article Snippet: Sections were then incubated with phalloidin mixture (1:1 000, Abcam) in the dark for 1 h at room temperature.

    Article Title: Maladaptive inflammatory signaling in old mice impairs colonic regeneration by promoting a sustained fetal-like epithelial state
    Article Snippet: RNA in situ hybridization for Lgr5 expression was conducted on 7 μm paraffin sections using the RNAscope 2.5 High Definition (HD) – Red Assay (Advanced Cell Diagnostics, 322350).

    Sample Prep:

    Article Title: Dorsal root ganglion-targeted analgesic delivery for effective relief of neuropathic pain
    Article Snippet: RNA-scope assay was performed for in situ evaluation of Adam8 (Advanced Cell Diagnostics, #1047421-C1) in the DRG of the mice using RNA-Scope 2.5 HD Red Kit (Advanced Cell Diagnostics, #322360) following our recent work [ , ].

    Article Title: Prdm16 regulates the postnatal fate of embryonic radial glia via Vcam1-dependent mechanisms
    Article Snippet: We then washed the whole-mounts 3 times with PBS with 0.1% Triton-X100, blocked in 10% normal donkey serum in PBS with 2% Triton-X100 at room temperature for 1 h, and stained with primary antibodies against Vcam1 (BD Biosciences, 550547, 1:100), γ-Tubulin (Sigma, T5192-25UL, 1:1000), and SOX2 (R&D Systems, AF2018, 1:1000) for 48 h at 4 °C.

    Article Title: Nr4a1 modulates inflammation and heart regeneration in zebrafish
    Article Snippet: The probe synthesis was conducted using the Invitrogen MEGAscript T7 Transcription Kit with the DIG RNA labeling mix (Roche).

    Article Title: mRNA-based in vivo induction of ETV2 to restore blood flow in a preclinical mouse hindlimb ischemia model
    Article Snippet: In situ hybridization (ISH) was performed using an RNAscope 2.5 LS Reagent Kit-BROWN (Advanced Cell Diagnostics [ACD], Hayward, CA, cat no. 322100) for use with BOND RX Stainer (Leica Biosystems, Buffalo Grove, IL) according to the manufacturer’s instructions.

    Article Title: Notum as a Crucial Regulator of Matrix Integrity in Dentinogenesis
    Article Snippet: In situ hybridization was performed with the RNAscope 2.5 High‐Definition Assay‐Brown (Advanced Cell Diagnostics, Newark, CA, USA) according to the user manual 322452 (FFPE sample preparation and pretreatment) and 322310 (RNAscope 2.5 High‐Definition Detection Reagent‐Brown user manual) provided by the manufacturer.

    Article Title: Osteoclast-independent osteocyte dendrite defects in mice bearing the osteogenesis imperfecta-causing Sp7 R342C mutation
    Article Snippet: Sections were then incubated with phalloidin mixture (1:1 000, Abcam) in the dark for 1 h at room temperature.

    Article Title: Maladaptive inflammatory signaling in old mice impairs colonic regeneration by promoting a sustained fetal-like epithelial state
    Article Snippet: RNA in situ hybridization for Lgr5 expression was conducted on 7 μm paraffin sections using the RNAscope 2.5 High Definition (HD) – Red Assay (Advanced Cell Diagnostics, 322350).

    RNA In Situ Hybridization:

    Article Title: Dorsal root ganglion-targeted analgesic delivery for effective relief of neuropathic pain
    Article Snippet: RNA-scope assay was performed for in situ evaluation of Adam8 (Advanced Cell Diagnostics, #1047421-C1) in the DRG of the mice using RNA-Scope 2.5 HD Red Kit (Advanced Cell Diagnostics, #322360) following our recent work [ , ].

    Article Title: Prdm16 regulates the postnatal fate of embryonic radial glia via Vcam1-dependent mechanisms
    Article Snippet: We then washed the whole-mounts 3 times with PBS with 0.1% Triton-X100, blocked in 10% normal donkey serum in PBS with 2% Triton-X100 at room temperature for 1 h, and stained with primary antibodies against Vcam1 (BD Biosciences, 550547, 1:100), γ-Tubulin (Sigma, T5192-25UL, 1:1000), and SOX2 (R&D Systems, AF2018, 1:1000) for 48 h at 4 °C.

    Article Title: Nr4a1 modulates inflammation and heart regeneration in zebrafish
    Article Snippet: The probe synthesis was conducted using the Invitrogen MEGAscript T7 Transcription Kit with the DIG RNA labeling mix (Roche).

    Article Title: mRNA-based in vivo induction of ETV2 to restore blood flow in a preclinical mouse hindlimb ischemia model
    Article Snippet: In situ hybridization (ISH) was performed using an RNAscope 2.5 LS Reagent Kit-BROWN (Advanced Cell Diagnostics [ACD], Hayward, CA, cat no. 322100) for use with BOND RX Stainer (Leica Biosystems, Buffalo Grove, IL) according to the manufacturer’s instructions.

    Article Title: Notum as a Crucial Regulator of Matrix Integrity in Dentinogenesis
    Article Snippet: In situ hybridization was performed with the RNAscope 2.5 High‐Definition Assay‐Brown (Advanced Cell Diagnostics, Newark, CA, USA) according to the user manual 322452 (FFPE sample preparation and pretreatment) and 322310 (RNAscope 2.5 High‐Definition Detection Reagent‐Brown user manual) provided by the manufacturer.

    Article Title: Osteoclast-independent osteocyte dendrite defects in mice bearing the osteogenesis imperfecta-causing Sp7 R342C mutation
    Article Snippet: Sections were then incubated with phalloidin mixture (1:1 000, Abcam) in the dark for 1 h at room temperature.

    Article Title: Maladaptive inflammatory signaling in old mice impairs colonic regeneration by promoting a sustained fetal-like epithelial state
    Article Snippet: RNA in situ hybridization for Lgr5 expression was conducted on 7 μm paraffin sections using the RNAscope 2.5 High Definition (HD) – Red Assay (Advanced Cell Diagnostics, 322350).

    Expressing:

    Article Title: Dorsal root ganglion-targeted analgesic delivery for effective relief of neuropathic pain
    Article Snippet: RNA-scope assay was performed for in situ evaluation of Adam8 (Advanced Cell Diagnostics, #1047421-C1) in the DRG of the mice using RNA-Scope 2.5 HD Red Kit (Advanced Cell Diagnostics, #322360) following our recent work [ , ].

    Article Title: Prdm16 regulates the postnatal fate of embryonic radial glia via Vcam1-dependent mechanisms
    Article Snippet: We then washed the whole-mounts 3 times with PBS with 0.1% Triton-X100, blocked in 10% normal donkey serum in PBS with 2% Triton-X100 at room temperature for 1 h, and stained with primary antibodies against Vcam1 (BD Biosciences, 550547, 1:100), γ-Tubulin (Sigma, T5192-25UL, 1:1000), and SOX2 (R&D Systems, AF2018, 1:1000) for 48 h at 4 °C.

    Article Title: Nr4a1 modulates inflammation and heart regeneration in zebrafish
    Article Snippet: The probe synthesis was conducted using the Invitrogen MEGAscript T7 Transcription Kit with the DIG RNA labeling mix (Roche).

    Article Title: mRNA-based in vivo induction of ETV2 to restore blood flow in a preclinical mouse hindlimb ischemia model
    Article Snippet: In situ hybridization (ISH) was performed using an RNAscope 2.5 LS Reagent Kit-BROWN (Advanced Cell Diagnostics [ACD], Hayward, CA, cat no. 322100) for use with BOND RX Stainer (Leica Biosystems, Buffalo Grove, IL) according to the manufacturer’s instructions.

    Article Title: Notum as a Crucial Regulator of Matrix Integrity in Dentinogenesis
    Article Snippet: In situ hybridization was performed with the RNAscope 2.5 High‐Definition Assay‐Brown (Advanced Cell Diagnostics, Newark, CA, USA) according to the user manual 322452 (FFPE sample preparation and pretreatment) and 322310 (RNAscope 2.5 High‐Definition Detection Reagent‐Brown user manual) provided by the manufacturer.

    Article Title: Osteoclast-independent osteocyte dendrite defects in mice bearing the osteogenesis imperfecta-causing Sp7 R342C mutation
    Article Snippet: Sections were then incubated with phalloidin mixture (1:1 000, Abcam) in the dark for 1 h at room temperature.

    Article Title: Maladaptive inflammatory signaling in old mice impairs colonic regeneration by promoting a sustained fetal-like epithelial state
    Article Snippet: RNA in situ hybridization for Lgr5 expression was conducted on 7 μm paraffin sections using the RNAscope 2.5 High Definition (HD) – Red Assay (Advanced Cell Diagnostics, 322350).



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    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 <t>RNAscope</t> 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.
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    Image Search Results


    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.

    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