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human p110 isoforms  (Santa Cruz Biotechnology)


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    Santa Cruz Biotechnology human p110 isoforms
    Figure 6. Lyn is essential to the activation of PI 3-kinase through phosphorylation of its <t>p110</t> subunit. (A; Supplementary movies M1–M4) HEK 293–TLR2 cells were transfected with a fusion protein combining GFP and the PH domain of AKT. Transfected cells were either co-transfected with LynDK (500 ng/ml) or incubated with PP2. Cells were then stimulated with Pam3 (100 ng/ml) and the kinetics of the AKT–PH construct was observed by microvideoscopy using Zeiss Axiovert inverted microscope equipped with the Metafluor imaging system. Presented here are the images corresponding to 15 min of Pam3 stimulation. Controls correspond to cells incubated with DMSO and pcDNA empty vector, or cells incubated with LY294002 (25 mM), a specific inhibitor of PI 3-kinase. (B) HEK 293–TLR2 cells were transfected with pcDNA vector (vehicle) or LynDK (500 ng/ml) and stimulated with Pam3 (100 ng/ml). Lysates were immunoprecipitated with anti-Flag Abs and recruitment of PI 3-kinase to TLR2 was observed by Western blot with anti-p85a Abs. (C) Tyrosine phosphorylation of the p85a subunit was evaluated by Western blot in HEK 293–TLR2 transfected with LynDK (500 ng/ml) and stimulated with Pam3 (100 ng/ml). Anti-phosphotyrosine (4G10 clone) and anti-p85a Abs were used for immuno- precipitation and Western blot. (D) HEK 293–TLR2 were transfected with LynDK (500 ng/ml) and stimulated with Pam3 (100 ng/ml) and lysates were immunoprecipitated with either 4G10 or anti-p110 Abs. Phosphorylation of p110 was then revealed by Western blot with anti-p110 or 4G10 Abs. Controls correspond to cells transfected with pcDNA empty vector. (E) THP1–CD14 cells were incubated with PP2 (25mM) or DMSO, stimulated with Pam3 (100 ng/ml) and lysed. Phosphorylation of p110 catalytic subunit of PI 3-kinase was revealed by Western blot of THP1–CD14 lysates immunoprecipitated with either 4G10 or anti-p110 Abs. Controls correspond to cells treated with DMSO. These results are representative of three independent experiments.
    Human P110 Isoforms, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 275 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human p110 isoforms/product/Santa Cruz Biotechnology
    Average 93 stars, based on 275 article reviews
    human p110 isoforms - by Bioz Stars, 2026-06
    93/100 stars

    Images

    1) Product Images from "Src-family-tyrosine kinase Lyn is critical for TLR2-mediated NF-κB activation through the PI 3-kinase signaling pathway."

    Article Title: Src-family-tyrosine kinase Lyn is critical for TLR2-mediated NF-κB activation through the PI 3-kinase signaling pathway.

    Journal: Innate immunity

    doi: 10.1177/1753425915586075

    Figure 6. Lyn is essential to the activation of PI 3-kinase through phosphorylation of its p110 subunit. (A; Supplementary movies M1–M4) HEK 293–TLR2 cells were transfected with a fusion protein combining GFP and the PH domain of AKT. Transfected cells were either co-transfected with LynDK (500 ng/ml) or incubated with PP2. Cells were then stimulated with Pam3 (100 ng/ml) and the kinetics of the AKT–PH construct was observed by microvideoscopy using Zeiss Axiovert inverted microscope equipped with the Metafluor imaging system. Presented here are the images corresponding to 15 min of Pam3 stimulation. Controls correspond to cells incubated with DMSO and pcDNA empty vector, or cells incubated with LY294002 (25 mM), a specific inhibitor of PI 3-kinase. (B) HEK 293–TLR2 cells were transfected with pcDNA vector (vehicle) or LynDK (500 ng/ml) and stimulated with Pam3 (100 ng/ml). Lysates were immunoprecipitated with anti-Flag Abs and recruitment of PI 3-kinase to TLR2 was observed by Western blot with anti-p85a Abs. (C) Tyrosine phosphorylation of the p85a subunit was evaluated by Western blot in HEK 293–TLR2 transfected with LynDK (500 ng/ml) and stimulated with Pam3 (100 ng/ml). Anti-phosphotyrosine (4G10 clone) and anti-p85a Abs were used for immuno- precipitation and Western blot. (D) HEK 293–TLR2 were transfected with LynDK (500 ng/ml) and stimulated with Pam3 (100 ng/ml) and lysates were immunoprecipitated with either 4G10 or anti-p110 Abs. Phosphorylation of p110 was then revealed by Western blot with anti-p110 or 4G10 Abs. Controls correspond to cells transfected with pcDNA empty vector. (E) THP1–CD14 cells were incubated with PP2 (25mM) or DMSO, stimulated with Pam3 (100 ng/ml) and lysed. Phosphorylation of p110 catalytic subunit of PI 3-kinase was revealed by Western blot of THP1–CD14 lysates immunoprecipitated with either 4G10 or anti-p110 Abs. Controls correspond to cells treated with DMSO. These results are representative of three independent experiments.
    Figure Legend Snippet: Figure 6. Lyn is essential to the activation of PI 3-kinase through phosphorylation of its p110 subunit. (A; Supplementary movies M1–M4) HEK 293–TLR2 cells were transfected with a fusion protein combining GFP and the PH domain of AKT. Transfected cells were either co-transfected with LynDK (500 ng/ml) or incubated with PP2. Cells were then stimulated with Pam3 (100 ng/ml) and the kinetics of the AKT–PH construct was observed by microvideoscopy using Zeiss Axiovert inverted microscope equipped with the Metafluor imaging system. Presented here are the images corresponding to 15 min of Pam3 stimulation. Controls correspond to cells incubated with DMSO and pcDNA empty vector, or cells incubated with LY294002 (25 mM), a specific inhibitor of PI 3-kinase. (B) HEK 293–TLR2 cells were transfected with pcDNA vector (vehicle) or LynDK (500 ng/ml) and stimulated with Pam3 (100 ng/ml). Lysates were immunoprecipitated with anti-Flag Abs and recruitment of PI 3-kinase to TLR2 was observed by Western blot with anti-p85a Abs. (C) Tyrosine phosphorylation of the p85a subunit was evaluated by Western blot in HEK 293–TLR2 transfected with LynDK (500 ng/ml) and stimulated with Pam3 (100 ng/ml). Anti-phosphotyrosine (4G10 clone) and anti-p85a Abs were used for immuno- precipitation and Western blot. (D) HEK 293–TLR2 were transfected with LynDK (500 ng/ml) and stimulated with Pam3 (100 ng/ml) and lysates were immunoprecipitated with either 4G10 or anti-p110 Abs. Phosphorylation of p110 was then revealed by Western blot with anti-p110 or 4G10 Abs. Controls correspond to cells transfected with pcDNA empty vector. (E) THP1–CD14 cells were incubated with PP2 (25mM) or DMSO, stimulated with Pam3 (100 ng/ml) and lysed. Phosphorylation of p110 catalytic subunit of PI 3-kinase was revealed by Western blot of THP1–CD14 lysates immunoprecipitated with either 4G10 or anti-p110 Abs. Controls correspond to cells treated with DMSO. These results are representative of three independent experiments.

    Techniques Used: Activation Assay, Phospho-proteomics, Transfection, Incubation, Construct, Inverted Microscopy, Imaging, Plasmid Preparation, Immunoprecipitation, Western Blot

    Figure 7. Lyn controls the PI 3-kinase pathway through phosphorylation of its p110 subunit after TLR2 engagement. The presence of bacteria-derived acylated lipoproteins results in the heterodimerization of TLR2 with TLR1/6 in membrane microdomains. MyD88/ IRAK are recruited to the receptor and lead to degradation of IkB and translocation of NF-kB subunits. A Rac/PI 3-kinase-dependent signaling pathway has also been described, including CD14 and Lyn that contribute to the activation cluster. After tyrosine phos- phorylation of the cytoplasmic domain of TLR2, the p85a subunit of PI 3-kinase is recruited to the receptor and phosphorylated on tyrosine. A Lyn-dependent tyrosine-phosphorylation is required to activate the PI 3-kinase catalytic subunit p110 that allows for recruitment of AKT to the inner membrane, precluding a cascade that results in transactivation of the p65 subunit of NF-kB. This leads to the nuclear translocation of a functional p50/p65 NF-kB heterodimer that results in the gene expression of pro-inflammatory cytokines.
    Figure Legend Snippet: Figure 7. Lyn controls the PI 3-kinase pathway through phosphorylation of its p110 subunit after TLR2 engagement. The presence of bacteria-derived acylated lipoproteins results in the heterodimerization of TLR2 with TLR1/6 in membrane microdomains. MyD88/ IRAK are recruited to the receptor and lead to degradation of IkB and translocation of NF-kB subunits. A Rac/PI 3-kinase-dependent signaling pathway has also been described, including CD14 and Lyn that contribute to the activation cluster. After tyrosine phos- phorylation of the cytoplasmic domain of TLR2, the p85a subunit of PI 3-kinase is recruited to the receptor and phosphorylated on tyrosine. A Lyn-dependent tyrosine-phosphorylation is required to activate the PI 3-kinase catalytic subunit p110 that allows for recruitment of AKT to the inner membrane, precluding a cascade that results in transactivation of the p65 subunit of NF-kB. This leads to the nuclear translocation of a functional p50/p65 NF-kB heterodimer that results in the gene expression of pro-inflammatory cytokines.

    Techniques Used: Phospho-proteomics, Bacteria, Derivative Assay, Membrane, Translocation Assay, Activation Assay, Functional Assay, Gene Expression



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    human adar1 p110 isoform antibody  (Proteintech)
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    Proteintech human adar1 p110 isoform antibody
    (A) Western blots showing the protein levels of <t>ADAR1</t> and PD-L1 in Capan1, HPAC, BT549 and MB-231 cells treated with the indicated concentrations of ATRA for 24 h. PDAC, pancreatic ductal adenocarcinoma; TNBC, triple-negative breast cancer. (B) Western blots showing the protein levels of Adar1 and Pd-l1 in Panc02 and 4T1 cells treated with the indicated doses of ATRA for 24 h. (C) Western blots showing exogenous MYC-GFP-tagged ADAR1 protein expression in HPAC or MB-231 cells treated with ATRA for 24 h. (D) Western blots showing exogenous FLAG-tagged ADAR1 protein expression in BxPC3 cells treated with ATRA for 24 h. (E) Upper, Western blots showing the protein levels of ADAR1 in HPAC cells treated with ATRA or MG132 alone or in combination for the indicated time. Lower, Quantification of relative ADAR1 expression. (F) Co-immunoprecipitation (Co- IP) blots showing polyubiquitination (Ub) and K48-linked ubiquitination (Ub-K48) of ADAR1 in BxPC3 and Capan1 cells exogenously expressing MYC-GFP-tagged ADAR1 and treated with ATRA. (G) Co-IP blots showing polyubiquitination and Ub-K48 of ADAR1 in HPAC cells treated with ATRA or MG132.
    Human Adar1 P110 Isoform Antibody, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human adar1 p110 isoform antibody/product/Proteintech
    Average 96 stars, based on 1 article reviews
    human adar1 p110 isoform antibody - by Bioz Stars, 2026-06
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    human p110 isoforms  (Santa Cruz Biotechnology)
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    Santa Cruz Biotechnology human p110 isoforms
    Figure 6. Lyn is essential to the activation of PI 3-kinase through phosphorylation of its <t>p110</t> subunit. (A; Supplementary movies M1–M4) HEK 293–TLR2 cells were transfected with a fusion protein combining GFP and the PH domain of AKT. Transfected cells were either co-transfected with LynDK (500 ng/ml) or incubated with PP2. Cells were then stimulated with Pam3 (100 ng/ml) and the kinetics of the AKT–PH construct was observed by microvideoscopy using Zeiss Axiovert inverted microscope equipped with the Metafluor imaging system. Presented here are the images corresponding to 15 min of Pam3 stimulation. Controls correspond to cells incubated with DMSO and pcDNA empty vector, or cells incubated with LY294002 (25 mM), a specific inhibitor of PI 3-kinase. (B) HEK 293–TLR2 cells were transfected with pcDNA vector (vehicle) or LynDK (500 ng/ml) and stimulated with Pam3 (100 ng/ml). Lysates were immunoprecipitated with anti-Flag Abs and recruitment of PI 3-kinase to TLR2 was observed by Western blot with anti-p85a Abs. (C) Tyrosine phosphorylation of the p85a subunit was evaluated by Western blot in HEK 293–TLR2 transfected with LynDK (500 ng/ml) and stimulated with Pam3 (100 ng/ml). Anti-phosphotyrosine (4G10 clone) and anti-p85a Abs were used for immuno- precipitation and Western blot. (D) HEK 293–TLR2 were transfected with LynDK (500 ng/ml) and stimulated with Pam3 (100 ng/ml) and lysates were immunoprecipitated with either 4G10 or anti-p110 Abs. Phosphorylation of p110 was then revealed by Western blot with anti-p110 or 4G10 Abs. Controls correspond to cells transfected with pcDNA empty vector. (E) THP1–CD14 cells were incubated with PP2 (25mM) or DMSO, stimulated with Pam3 (100 ng/ml) and lysed. Phosphorylation of p110 catalytic subunit of PI 3-kinase was revealed by Western blot of THP1–CD14 lysates immunoprecipitated with either 4G10 or anti-p110 Abs. Controls correspond to cells treated with DMSO. These results are representative of three independent experiments.
    Human P110 Isoforms, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human p110 isoforms/product/Santa Cruz Biotechnology
    Average 93 stars, based on 1 article reviews
    human p110 isoforms - by Bioz Stars, 2026-06
    93/100 stars
    		  Buy from Supplier

    Image Search Results


    (A) Western blots showing the protein levels of ADAR1 and PD-L1 in Capan1, HPAC, BT549 and MB-231 cells treated with the indicated concentrations of ATRA for 24 h. PDAC, pancreatic ductal adenocarcinoma; TNBC, triple-negative breast cancer. (B) Western blots showing the protein levels of Adar1 and Pd-l1 in Panc02 and 4T1 cells treated with the indicated doses of ATRA for 24 h. (C) Western blots showing exogenous MYC-GFP-tagged ADAR1 protein expression in HPAC or MB-231 cells treated with ATRA for 24 h. (D) Western blots showing exogenous FLAG-tagged ADAR1 protein expression in BxPC3 cells treated with ATRA for 24 h. (E) Upper, Western blots showing the protein levels of ADAR1 in HPAC cells treated with ATRA or MG132 alone or in combination for the indicated time. Lower, Quantification of relative ADAR1 expression. (F) Co-immunoprecipitation (Co- IP) blots showing polyubiquitination (Ub) and K48-linked ubiquitination (Ub-K48) of ADAR1 in BxPC3 and Capan1 cells exogenously expressing MYC-GFP-tagged ADAR1 and treated with ATRA. (G) Co-IP blots showing polyubiquitination and Ub-K48 of ADAR1 in HPAC cells treated with ATRA or MG132.

    Journal: bioRxiv

    Article Title: All-trans retinoic acid-mediated ADAR1 degradation synergizes with PD-1 blockade to suppress pancreatic cancer

    doi: 10.1101/2024.10.20.619300

    Figure Lengend Snippet: (A) Western blots showing the protein levels of ADAR1 and PD-L1 in Capan1, HPAC, BT549 and MB-231 cells treated with the indicated concentrations of ATRA for 24 h. PDAC, pancreatic ductal adenocarcinoma; TNBC, triple-negative breast cancer. (B) Western blots showing the protein levels of Adar1 and Pd-l1 in Panc02 and 4T1 cells treated with the indicated doses of ATRA for 24 h. (C) Western blots showing exogenous MYC-GFP-tagged ADAR1 protein expression in HPAC or MB-231 cells treated with ATRA for 24 h. (D) Western blots showing exogenous FLAG-tagged ADAR1 protein expression in BxPC3 cells treated with ATRA for 24 h. (E) Upper, Western blots showing the protein levels of ADAR1 in HPAC cells treated with ATRA or MG132 alone or in combination for the indicated time. Lower, Quantification of relative ADAR1 expression. (F) Co-immunoprecipitation (Co- IP) blots showing polyubiquitination (Ub) and K48-linked ubiquitination (Ub-K48) of ADAR1 in BxPC3 and Capan1 cells exogenously expressing MYC-GFP-tagged ADAR1 and treated with ATRA. (G) Co-IP blots showing polyubiquitination and Ub-K48 of ADAR1 in HPAC cells treated with ATRA or MG132.

    Article Snippet: Human ADAR1 (p150 isoform) antibody was purchased from Bethyl Laboratories, Inc. Human ADAR1 (p110 isoform) antibody and vinculin antibody were purchased from Proteintech.

    Techniques: Western Blot, Expressing, Immunoprecipitation, Co-Immunoprecipitation Assay

    (A) Schematic representation of the treatment schedule in the Panc02 mouse tumor model. Panc02 tumor-bearing mice were given ATRA or placebo pellets (10 mg per mouse, subcutaneously [s.c.], 21-day release). Administration of anti-PD-1 antibody or IgG control (100 μg per mouse, intraperitoneally [i.p.], 3 times weekly) was started along with ATRA or placebo pellets. Tumor growth, body weight, and mouse survival were measured. (B) Left, representative images showing Panc02 tumors harvested from mice that received the indicated treatments. Scale bar, 1.2 cm. Right, tumor weight. Data represent mean ± SD. (C) Tumor growth curves of individual mice injected with Panc02 tumor cells and subjected to the indicated treatments. (D) Mean tumor growth in mice injected with Panc02 tumor cells and subjected to the indicated treatments. Each dot represents the mean tumor size from 9 mice in each treatment group; whiskers represent SD. (E) Survival curves of mice that received the indicated treatments (9 mice/group). (F) Survival curves of KPC mice that received the indicated treatments (3 mice/group). (G) Western blots showing Adar1 and Pd-l1 protein expression levels in Panc02 cells from mice treated with the indicated regimens. (H) Body weight of tumor-bearing mice treated with the indicated treatments. Data are shown as mean ± SD. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001; ns, not significant.

    Journal: bioRxiv

    Article Title: All-trans retinoic acid-mediated ADAR1 degradation synergizes with PD-1 blockade to suppress pancreatic cancer

    doi: 10.1101/2024.10.20.619300

    Figure Lengend Snippet: (A) Schematic representation of the treatment schedule in the Panc02 mouse tumor model. Panc02 tumor-bearing mice were given ATRA or placebo pellets (10 mg per mouse, subcutaneously [s.c.], 21-day release). Administration of anti-PD-1 antibody or IgG control (100 μg per mouse, intraperitoneally [i.p.], 3 times weekly) was started along with ATRA or placebo pellets. Tumor growth, body weight, and mouse survival were measured. (B) Left, representative images showing Panc02 tumors harvested from mice that received the indicated treatments. Scale bar, 1.2 cm. Right, tumor weight. Data represent mean ± SD. (C) Tumor growth curves of individual mice injected with Panc02 tumor cells and subjected to the indicated treatments. (D) Mean tumor growth in mice injected with Panc02 tumor cells and subjected to the indicated treatments. Each dot represents the mean tumor size from 9 mice in each treatment group; whiskers represent SD. (E) Survival curves of mice that received the indicated treatments (9 mice/group). (F) Survival curves of KPC mice that received the indicated treatments (3 mice/group). (G) Western blots showing Adar1 and Pd-l1 protein expression levels in Panc02 cells from mice treated with the indicated regimens. (H) Body weight of tumor-bearing mice treated with the indicated treatments. Data are shown as mean ± SD. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001; ns, not significant.

    Article Snippet: Human ADAR1 (p150 isoform) antibody was purchased from Bethyl Laboratories, Inc. Human ADAR1 (p110 isoform) antibody and vinculin antibody were purchased from Proteintech.

    Techniques: Control, Injection, Western Blot, Expressing

    (A) Heatmap showing differential marker expression in CD45+ tumor-infiltrating lymphocyte (TIL) clusters identified by analysis of time-of-flight mass cytometry data with viSNE and FlowSOM. DC, dendritic cell; MDSC, myeloid-derived suppressor cell; Treg, regulatory T cell; NK, natural killer cell. (B) Annotation of TIL populations based on differential marker expression as shown in the heatmap (A) and Figure S8. TAM, tumor-associated macrophage. (C) Cellular distribution and clustering as defined by tSNE1 and tSNE2, colored by cell phenotype, for Panc02 tumors subjected to the indicated treatments. Data show all normalized viable single cells; analysis was conducted using viSNE and FlowSOM. (D) Percentage of CD8+ T cells among total CD45+ TILs under the indicated treatments. (E) T-cell cytotoxicity (CCK-8) showing effects of the indicated treatments on pancreatic cancer cell viability. Target Panc28 cancer cells were engineered to express Fcγ receptor 2A fused with the luciferase Luc2 at the C- terminus (Panc28-RL2), and then incubated with CD8+ T cells and the CD3 antibody OKT3 for 3 days. Data represent mean ± SD. P values were calculated by Student t test. (F) T-cell cytotoxicity (CCK-8) assay showing effects of ATRA on pancreatic cancer cell viability. Panc28-RL2 cells overexpressing MYC-GFP-ADAR1 or MYC-GFP (control) were incubated with CD8+ T cells and the CD3 antibody OKT3 for 3 days. Data represent mean ± SD. P values calculated by Student t test.

    Journal: bioRxiv

    Article Title: All-trans retinoic acid-mediated ADAR1 degradation synergizes with PD-1 blockade to suppress pancreatic cancer

    doi: 10.1101/2024.10.20.619300

    Figure Lengend Snippet: (A) Heatmap showing differential marker expression in CD45+ tumor-infiltrating lymphocyte (TIL) clusters identified by analysis of time-of-flight mass cytometry data with viSNE and FlowSOM. DC, dendritic cell; MDSC, myeloid-derived suppressor cell; Treg, regulatory T cell; NK, natural killer cell. (B) Annotation of TIL populations based on differential marker expression as shown in the heatmap (A) and Figure S8. TAM, tumor-associated macrophage. (C) Cellular distribution and clustering as defined by tSNE1 and tSNE2, colored by cell phenotype, for Panc02 tumors subjected to the indicated treatments. Data show all normalized viable single cells; analysis was conducted using viSNE and FlowSOM. (D) Percentage of CD8+ T cells among total CD45+ TILs under the indicated treatments. (E) T-cell cytotoxicity (CCK-8) showing effects of the indicated treatments on pancreatic cancer cell viability. Target Panc28 cancer cells were engineered to express Fcγ receptor 2A fused with the luciferase Luc2 at the C- terminus (Panc28-RL2), and then incubated with CD8+ T cells and the CD3 antibody OKT3 for 3 days. Data represent mean ± SD. P values were calculated by Student t test. (F) T-cell cytotoxicity (CCK-8) assay showing effects of ATRA on pancreatic cancer cell viability. Panc28-RL2 cells overexpressing MYC-GFP-ADAR1 or MYC-GFP (control) were incubated with CD8+ T cells and the CD3 antibody OKT3 for 3 days. Data represent mean ± SD. P values calculated by Student t test.

    Article Snippet: Human ADAR1 (p150 isoform) antibody was purchased from Bethyl Laboratories, Inc. Human ADAR1 (p110 isoform) antibody and vinculin antibody were purchased from Proteintech.

    Techniques: Marker, Expressing, Mass Cytometry, Derivative Assay, CCK-8 Assay, Luciferase, Incubation, Control

    (A) ADAR gene expression profiling by GEPIA2. Red dots: tumor tissues; green dots: normal tissues. The bars represent the median expression of certain tumor type or normal tissue. BRCA, invasive breast carcinoma; ESCA, esophageal carcinoma; HNSC, head and neck squamous cell carcinoma; LGG, lower-grade brain glioma; LIHC, liver hepatocellular carcinoma; PAAD, pancreatic adenocarcinoma; SKCM, skin cutaneous melanoma; STAD, stomach adenocarcinoma; TGCT, testicular germ cell tumor. (B) Kaplan-Meier overall survival curves of 135 pancreatic cancer patients in the cohort with high and low ADAR1 expression determined by immunohistochemical staining. (C) Representative images of the immunohistochemical staining of ADAR1 in tumor tissue from patients in the MDA cohort. Scale bars, 50 μm. (D) Survival curves of breast cancer patients from 2 independent cohorts by ADAR1 expression level. (i) Liu et al. , 2014; (ii) TCGA. (E) Representative images of the immunohistochemical staining of ADAR1 and PD-L1 in tumor tissue from pancreatic cancer patients before treatment with ATRA and nivolumab. Scale bars, 100 μm. (F) Kaplan-Meier overall survival curves for 9 pancreatic cancer patients treated with ATRA 45 mg/m 2 /day plus nivolumab or ATRA ≥60 mg/m 2 /day plus nivolumab. (G) Schematic illustration of the mechanism of switching “cold” (immune-excluded) tumors to “hot” (immune-infiltrated) tumors by using ATRA and anti-PD-1 antibody.

    Journal: bioRxiv

    Article Title: All-trans retinoic acid-mediated ADAR1 degradation synergizes with PD-1 blockade to suppress pancreatic cancer

    doi: 10.1101/2024.10.20.619300

    Figure Lengend Snippet: (A) ADAR gene expression profiling by GEPIA2. Red dots: tumor tissues; green dots: normal tissues. The bars represent the median expression of certain tumor type or normal tissue. BRCA, invasive breast carcinoma; ESCA, esophageal carcinoma; HNSC, head and neck squamous cell carcinoma; LGG, lower-grade brain glioma; LIHC, liver hepatocellular carcinoma; PAAD, pancreatic adenocarcinoma; SKCM, skin cutaneous melanoma; STAD, stomach adenocarcinoma; TGCT, testicular germ cell tumor. (B) Kaplan-Meier overall survival curves of 135 pancreatic cancer patients in the cohort with high and low ADAR1 expression determined by immunohistochemical staining. (C) Representative images of the immunohistochemical staining of ADAR1 in tumor tissue from patients in the MDA cohort. Scale bars, 50 μm. (D) Survival curves of breast cancer patients from 2 independent cohorts by ADAR1 expression level. (i) Liu et al. , 2014; (ii) TCGA. (E) Representative images of the immunohistochemical staining of ADAR1 and PD-L1 in tumor tissue from pancreatic cancer patients before treatment with ATRA and nivolumab. Scale bars, 100 μm. (F) Kaplan-Meier overall survival curves for 9 pancreatic cancer patients treated with ATRA 45 mg/m 2 /day plus nivolumab or ATRA ≥60 mg/m 2 /day plus nivolumab. (G) Schematic illustration of the mechanism of switching “cold” (immune-excluded) tumors to “hot” (immune-infiltrated) tumors by using ATRA and anti-PD-1 antibody.

    Article Snippet: Human ADAR1 (p150 isoform) antibody was purchased from Bethyl Laboratories, Inc. Human ADAR1 (p110 isoform) antibody and vinculin antibody were purchased from Proteintech.

    Techniques: Expressing, Immunohistochemical staining, Staining

    Figure 6. Lyn is essential to the activation of PI 3-kinase through phosphorylation of its p110 subunit. (A; Supplementary movies M1–M4) HEK 293–TLR2 cells were transfected with a fusion protein combining GFP and the PH domain of AKT. Transfected cells were either co-transfected with LynDK (500 ng/ml) or incubated with PP2. Cells were then stimulated with Pam3 (100 ng/ml) and the kinetics of the AKT–PH construct was observed by microvideoscopy using Zeiss Axiovert inverted microscope equipped with the Metafluor imaging system. Presented here are the images corresponding to 15 min of Pam3 stimulation. Controls correspond to cells incubated with DMSO and pcDNA empty vector, or cells incubated with LY294002 (25 mM), a specific inhibitor of PI 3-kinase. (B) HEK 293–TLR2 cells were transfected with pcDNA vector (vehicle) or LynDK (500 ng/ml) and stimulated with Pam3 (100 ng/ml). Lysates were immunoprecipitated with anti-Flag Abs and recruitment of PI 3-kinase to TLR2 was observed by Western blot with anti-p85a Abs. (C) Tyrosine phosphorylation of the p85a subunit was evaluated by Western blot in HEK 293–TLR2 transfected with LynDK (500 ng/ml) and stimulated with Pam3 (100 ng/ml). Anti-phosphotyrosine (4G10 clone) and anti-p85a Abs were used for immuno- precipitation and Western blot. (D) HEK 293–TLR2 were transfected with LynDK (500 ng/ml) and stimulated with Pam3 (100 ng/ml) and lysates were immunoprecipitated with either 4G10 or anti-p110 Abs. Phosphorylation of p110 was then revealed by Western blot with anti-p110 or 4G10 Abs. Controls correspond to cells transfected with pcDNA empty vector. (E) THP1–CD14 cells were incubated with PP2 (25mM) or DMSO, stimulated with Pam3 (100 ng/ml) and lysed. Phosphorylation of p110 catalytic subunit of PI 3-kinase was revealed by Western blot of THP1–CD14 lysates immunoprecipitated with either 4G10 or anti-p110 Abs. Controls correspond to cells treated with DMSO. These results are representative of three independent experiments.

    Journal: Innate immunity

    Article Title: Src-family-tyrosine kinase Lyn is critical for TLR2-mediated NF-κB activation through the PI 3-kinase signaling pathway.

    doi: 10.1177/1753425915586075

    Figure Lengend Snippet: Figure 6. Lyn is essential to the activation of PI 3-kinase through phosphorylation of its p110 subunit. (A; Supplementary movies M1–M4) HEK 293–TLR2 cells were transfected with a fusion protein combining GFP and the PH domain of AKT. Transfected cells were either co-transfected with LynDK (500 ng/ml) or incubated with PP2. Cells were then stimulated with Pam3 (100 ng/ml) and the kinetics of the AKT–PH construct was observed by microvideoscopy using Zeiss Axiovert inverted microscope equipped with the Metafluor imaging system. Presented here are the images corresponding to 15 min of Pam3 stimulation. Controls correspond to cells incubated with DMSO and pcDNA empty vector, or cells incubated with LY294002 (25 mM), a specific inhibitor of PI 3-kinase. (B) HEK 293–TLR2 cells were transfected with pcDNA vector (vehicle) or LynDK (500 ng/ml) and stimulated with Pam3 (100 ng/ml). Lysates were immunoprecipitated with anti-Flag Abs and recruitment of PI 3-kinase to TLR2 was observed by Western blot with anti-p85a Abs. (C) Tyrosine phosphorylation of the p85a subunit was evaluated by Western blot in HEK 293–TLR2 transfected with LynDK (500 ng/ml) and stimulated with Pam3 (100 ng/ml). Anti-phosphotyrosine (4G10 clone) and anti-p85a Abs were used for immuno- precipitation and Western blot. (D) HEK 293–TLR2 were transfected with LynDK (500 ng/ml) and stimulated with Pam3 (100 ng/ml) and lysates were immunoprecipitated with either 4G10 or anti-p110 Abs. Phosphorylation of p110 was then revealed by Western blot with anti-p110 or 4G10 Abs. Controls correspond to cells transfected with pcDNA empty vector. (E) THP1–CD14 cells were incubated with PP2 (25mM) or DMSO, stimulated with Pam3 (100 ng/ml) and lysed. Phosphorylation of p110 catalytic subunit of PI 3-kinase was revealed by Western blot of THP1–CD14 lysates immunoprecipitated with either 4G10 or anti-p110 Abs. Controls correspond to cells treated with DMSO. These results are representative of three independent experiments.

    Article Snippet: Polyclonal Abs to phospho-AKT (Ser473), AKT, phospho-P65 (Ser536), P65, phosphoP38, phospho-ERK, phospho-SAP-JNK, ERK, SAPJNK, IkB and mAb P38 were from Cell Signaling (Danvers, MA, USA). mAb to Flag was from Sigma. mAbs against CD14 and aminoacid 800-1139 of human p110 isoforms were from Santa Cruz Biotechnology.

    Techniques: Activation Assay, Phospho-proteomics, Transfection, Incubation, Construct, Inverted Microscopy, Imaging, Plasmid Preparation, Immunoprecipitation, Western Blot

    Figure 7. Lyn controls the PI 3-kinase pathway through phosphorylation of its p110 subunit after TLR2 engagement. The presence of bacteria-derived acylated lipoproteins results in the heterodimerization of TLR2 with TLR1/6 in membrane microdomains. MyD88/ IRAK are recruited to the receptor and lead to degradation of IkB and translocation of NF-kB subunits. A Rac/PI 3-kinase-dependent signaling pathway has also been described, including CD14 and Lyn that contribute to the activation cluster. After tyrosine phos- phorylation of the cytoplasmic domain of TLR2, the p85a subunit of PI 3-kinase is recruited to the receptor and phosphorylated on tyrosine. A Lyn-dependent tyrosine-phosphorylation is required to activate the PI 3-kinase catalytic subunit p110 that allows for recruitment of AKT to the inner membrane, precluding a cascade that results in transactivation of the p65 subunit of NF-kB. This leads to the nuclear translocation of a functional p50/p65 NF-kB heterodimer that results in the gene expression of pro-inflammatory cytokines.

    Journal: Innate immunity

    Article Title: Src-family-tyrosine kinase Lyn is critical for TLR2-mediated NF-κB activation through the PI 3-kinase signaling pathway.

    doi: 10.1177/1753425915586075

    Figure Lengend Snippet: Figure 7. Lyn controls the PI 3-kinase pathway through phosphorylation of its p110 subunit after TLR2 engagement. The presence of bacteria-derived acylated lipoproteins results in the heterodimerization of TLR2 with TLR1/6 in membrane microdomains. MyD88/ IRAK are recruited to the receptor and lead to degradation of IkB and translocation of NF-kB subunits. A Rac/PI 3-kinase-dependent signaling pathway has also been described, including CD14 and Lyn that contribute to the activation cluster. After tyrosine phos- phorylation of the cytoplasmic domain of TLR2, the p85a subunit of PI 3-kinase is recruited to the receptor and phosphorylated on tyrosine. A Lyn-dependent tyrosine-phosphorylation is required to activate the PI 3-kinase catalytic subunit p110 that allows for recruitment of AKT to the inner membrane, precluding a cascade that results in transactivation of the p65 subunit of NF-kB. This leads to the nuclear translocation of a functional p50/p65 NF-kB heterodimer that results in the gene expression of pro-inflammatory cytokines.

    Article Snippet: Polyclonal Abs to phospho-AKT (Ser473), AKT, phospho-P65 (Ser536), P65, phosphoP38, phospho-ERK, phospho-SAP-JNK, ERK, SAPJNK, IkB and mAb P38 were from Cell Signaling (Danvers, MA, USA). mAb to Flag was from Sigma. mAbs against CD14 and aminoacid 800-1139 of human p110 isoforms were from Santa Cruz Biotechnology.

    Techniques: Phospho-proteomics, Bacteria, Derivative Assay, Membrane, Translocation Assay, Activation Assay, Functional Assay, Gene Expression