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apc mouse  (R&D Systems)


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    Structured Review

    R&D Systems apc mouse
    Apc Mouse, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/mica+b/Human+MICA%2FB+APC-conjugated+Antibody/10__1016_slash_j__omton__2026__201214-171-17-20
    Average 94 stars, based on 8 article reviews
    apc mouse - by Bioz Stars, 2026-10
    94/100 stars

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    Related Articles

    Fluorescence:

    Article Title: Metformin sensitizes leukemic cells to cytotoxic lymphocytes by increasing expression of intercellular adhesion molecule-1 (ICAM-1)
    Article Snippet: Metformin was from Santa Cruz Technologies. .. Fluorescence coupled antibodies against human ICAM-1 (AF1730), ULBP-1 (MAB1380), MICA/B (MAB13001) and the blocking antibody against NKG2D (MAB139) were from R&D Systems; CD33, HLA and CD138 were from Beckman; DR4 (12-6644-73), DR5 (12-9908-42) and Fas (BMS140FI) were from eBiosciences and 7AAD from BD Biosciences. ..

    Article Title: Metformin Sensitizes Leukemic Cells to Cytotoxic Lymphocytes by Increasing Expression of Intercellular Adhesion Molecule-1 (ICAM-1)
    Article Snippet: Metformin was from Santa Cruz Technologies. .. Fluorescence coupled antibodies against human ICAM-1 (AF1730), ULBP-1 (MAB1380), MICA/B (MAB13001) and the blocking antibody against NKG2D (MAB139) were from R&D Systems; CD33, HLA and CD138 were from Beckman; DR4 (12-6644-73), DR5 (12-9908-42) and Fas (BMS140FI) were from eBiosciences and 7AAD from BD Biosciences. .. Recombinant human Fas-Fc chimera (AFA0114121) was purchased from R&D Systems and human TRAIL-blocking antibody RIK2 (550912) from BD Biosciences and 7AAD from BD Biosciences.

    Article Title: Metformin Sensitizes Leukemic Cells to Cytotoxic Lymphocytes by Increasing Expression of Intercellular Adhesion Molecule-1 (ICAM-1)
    Article Snippet: Reagents and antibodies Metformin was from Santa Cruz Technologies. .. Fluorescence coupled antibodies against human ICAM-1 (AF1730), ULBP-1 (MAB1380), MICA/B (MAB13001) and the blocking antibody against NKG2D (MAB139) were from R&D Systems; CD33, HLA and CD138 were from Beckman; DR4 (12-6644-73), DR5 (12-9908- 42) and Fas (BMS140FI) were from eBiosciences and 7AAD from BD Biosciences. ..

    Blocking Assay:

    Article Title: Metformin sensitizes leukemic cells to cytotoxic lymphocytes by increasing expression of intercellular adhesion molecule-1 (ICAM-1)
    Article Snippet: Metformin was from Santa Cruz Technologies. .. Fluorescence coupled antibodies against human ICAM-1 (AF1730), ULBP-1 (MAB1380), MICA/B (MAB13001) and the blocking antibody against NKG2D (MAB139) were from R&D Systems; CD33, HLA and CD138 were from Beckman; DR4 (12-6644-73), DR5 (12-9908-42) and Fas (BMS140FI) were from eBiosciences and 7AAD from BD Biosciences. ..

    Article Title: Metformin Sensitizes Leukemic Cells to Cytotoxic Lymphocytes by Increasing Expression of Intercellular Adhesion Molecule-1 (ICAM-1)
    Article Snippet: Metformin was from Santa Cruz Technologies. .. Fluorescence coupled antibodies against human ICAM-1 (AF1730), ULBP-1 (MAB1380), MICA/B (MAB13001) and the blocking antibody against NKG2D (MAB139) were from R&D Systems; CD33, HLA and CD138 were from Beckman; DR4 (12-6644-73), DR5 (12-9908-42) and Fas (BMS140FI) were from eBiosciences and 7AAD from BD Biosciences. .. Recombinant human Fas-Fc chimera (AFA0114121) was purchased from R&D Systems and human TRAIL-blocking antibody RIK2 (550912) from BD Biosciences and 7AAD from BD Biosciences.

    Article Title: Metformin Sensitizes Leukemic Cells to Cytotoxic Lymphocytes by Increasing Expression of Intercellular Adhesion Molecule-1 (ICAM-1)
    Article Snippet: Reagents and antibodies Metformin was from Santa Cruz Technologies. .. Fluorescence coupled antibodies against human ICAM-1 (AF1730), ULBP-1 (MAB1380), MICA/B (MAB13001) and the blocking antibody against NKG2D (MAB139) were from R&D Systems; CD33, HLA and CD138 were from Beckman; DR4 (12-6644-73), DR5 (12-9908- 42) and Fas (BMS140FI) were from eBiosciences and 7AAD from BD Biosciences. ..

    Incubation:

    Article Title: Nano-Engineered Disruption of Heat shock protein 90 (Hsp90) Targets Drug-Induced Resistance and Relieves Natural Killer Cell Suppression in Breast Cancer
    Article Snippet: .. Cells were incubated with fluorescently labeled antibodies for NKG2D, CD158a, NKB1, CD244, KLRG1 (BioLegend, San Diego, CA), or MICA/B (R&D Systems, Minneapolis, MN) overnight at 4 °C and analyzed (C6 Accuri cyomteters Inc. Ann Arbor, MI). .. Data analysis was performed using FlowJo software (Tree Star Inc., Ashland OR) and Accuri cFlow plus software to obtain and confirm mean fluorescent intensity ( GNU.org ).

    Article Title: Nano-Engineered Disruption of Heat shock protein 90 (Hsp90) Targets Drug-Induced Resistance and Relieves Natural Killer Cell Suppression in Breast Cancer
    Article Snippet: .. Protein samples were resolved by SDS-PAGE and transferred to PVDF membranes prior to incubation at 4°C with indicated primary antibodies, mTOR and pMTOR Ser2448 , pAKT Thr308 and AKT, Phospho-p44/42 MAPK (Erk1/2) Thr202/Tyr204 , p44/42 MAPK (Erk1/2) pPRAS40 Thr246 , pSTAT3 Tyr705 , STAT3, PRAS40 and β-Actin antibodies were purchased from Cell Signaling Technology, pHck Tyr410 (Thermo Fisher Scientific) MICA/B (R&D Systems, Minneapolis, MN) and HSF-1 and HSF-1 Ser326 (abcam). ..

    Article Title: Nano-Engineered Disruption of Heat shock protein 90 (Hsp90) Targets Drug-Induced Resistance and Relieves Natural Killer Cell Suppression in Breast Cancer
    Article Snippet: .. Immunoblotting Protein samples were resolved by SDS-PAGE and transferred to PVDF membranes prior to incubation at 4°C with indicated primary antibodies, mTOR and pMTOR Ser2448 , pAKT Thr308 and AKT, Phospho-p44/42 MAPK (Erk1/2) Thr202/Tyr204 , p44/42 MAPK (Erk1/2) pPRAS40 Thr246 , pSTAT3 Tyr705 , STAT3, PRAS40 and β-Actin antibodies were purchased from Cell Signaling Technology, pHck Tyr410 (Thermo Fisher Scientific) MICA/B (R&D Systems, Minneapolis, MN) and HSF-1 and HSF-1 Ser326 (abcam). ..

    Labeling:

    Article Title: Nano-Engineered Disruption of Heat shock protein 90 (Hsp90) Targets Drug-Induced Resistance and Relieves Natural Killer Cell Suppression in Breast Cancer
    Article Snippet: .. Cells were incubated with fluorescently labeled antibodies for NKG2D, CD158a, NKB1, CD244, KLRG1 (BioLegend, San Diego, CA), or MICA/B (R&D Systems, Minneapolis, MN) overnight at 4 °C and analyzed (C6 Accuri cyomteters Inc. Ann Arbor, MI). .. Data analysis was performed using FlowJo software (Tree Star Inc., Ashland OR) and Accuri cFlow plus software to obtain and confirm mean fluorescent intensity ( GNU.org ).

    Article Title: Reducing fratricide of immune cells expressing NKG2D-based receptors
    Article Snippet: These results indicate that target-driven fratricide can be overcome using different approaches that can enable the development of T cell therapies where self-expression of the target ligand is a limiting factor. .. Materials and Methods Antibodies and Flow Cytometry Cells were stained with fluorochrome labeled CD3 (BD, 345766), CD4 (BD, 345809), CD8 (BD, 345772), CD314 (BD, 558071), CD45RA (BD, 550855), CD62L (BD, 555544), CD279 (eBioscience, 12-2799-42), CD19 (BD, 345791, CD223 (eBioscience, 25-2239-41), MICA/B (R&D Systems, FAB13001G-100), MICB (R&D Systems, FAB1599G), ULBP1 (R&D Systems, FAB1380C), ULBP2/5/6 (R&D Systems, FAB1298A), ULBP3 (R&D Systems, FAB1517P), ULBP4 (R&D Systems: FAB6285A), and corresponding isotypes according to standard protocols. .. Briefly, cells were harvested and resuspended in a buffer containing DPBS (Life Technologies, A1285801) supplemented with 5% human serum albumin (Octapharma, 68209-633-02) and 0.01% NaN3 (Sigma, S2002).

    SDS Page:

    Article Title: Nano-Engineered Disruption of Heat shock protein 90 (Hsp90) Targets Drug-Induced Resistance and Relieves Natural Killer Cell Suppression in Breast Cancer
    Article Snippet: .. Protein samples were resolved by SDS-PAGE and transferred to PVDF membranes prior to incubation at 4°C with indicated primary antibodies, mTOR and pMTOR Ser2448 , pAKT Thr308 and AKT, Phospho-p44/42 MAPK (Erk1/2) Thr202/Tyr204 , p44/42 MAPK (Erk1/2) pPRAS40 Thr246 , pSTAT3 Tyr705 , STAT3, PRAS40 and β-Actin antibodies were purchased from Cell Signaling Technology, pHck Tyr410 (Thermo Fisher Scientific) MICA/B (R&D Systems, Minneapolis, MN) and HSF-1 and HSF-1 Ser326 (abcam). ..

    Article Title: Nano-Engineered Disruption of Heat shock protein 90 (Hsp90) Targets Drug-Induced Resistance and Relieves Natural Killer Cell Suppression in Breast Cancer
    Article Snippet: .. Immunoblotting Protein samples were resolved by SDS-PAGE and transferred to PVDF membranes prior to incubation at 4°C with indicated primary antibodies, mTOR and pMTOR Ser2448 , pAKT Thr308 and AKT, Phospho-p44/42 MAPK (Erk1/2) Thr202/Tyr204 , p44/42 MAPK (Erk1/2) pPRAS40 Thr246 , pSTAT3 Tyr705 , STAT3, PRAS40 and β-Actin antibodies were purchased from Cell Signaling Technology, pHck Tyr410 (Thermo Fisher Scientific) MICA/B (R&D Systems, Minneapolis, MN) and HSF-1 and HSF-1 Ser326 (abcam). ..

    Flow Cytometry:

    Article Title: Reducing fratricide of immune cells expressing NKG2D-based receptors
    Article Snippet: These results indicate that target-driven fratricide can be overcome using different approaches that can enable the development of T cell therapies where self-expression of the target ligand is a limiting factor. .. Materials and Methods Antibodies and Flow Cytometry Cells were stained with fluorochrome labeled CD3 (BD, 345766), CD4 (BD, 345809), CD8 (BD, 345772), CD314 (BD, 558071), CD45RA (BD, 550855), CD62L (BD, 555544), CD279 (eBioscience, 12-2799-42), CD19 (BD, 345791, CD223 (eBioscience, 25-2239-41), MICA/B (R&D Systems, FAB13001G-100), MICB (R&D Systems, FAB1599G), ULBP1 (R&D Systems, FAB1380C), ULBP2/5/6 (R&D Systems, FAB1298A), ULBP3 (R&D Systems, FAB1517P), ULBP4 (R&D Systems: FAB6285A), and corresponding isotypes according to standard protocols. .. Briefly, cells were harvested and resuspended in a buffer containing DPBS (Life Technologies, A1285801) supplemented with 5% human serum albumin (Octapharma, 68209-633-02) and 0.01% NaN3 (Sigma, S2002).

    Staining:

    Article Title: Reducing fratricide of immune cells expressing NKG2D-based receptors
    Article Snippet: These results indicate that target-driven fratricide can be overcome using different approaches that can enable the development of T cell therapies where self-expression of the target ligand is a limiting factor. .. Materials and Methods Antibodies and Flow Cytometry Cells were stained with fluorochrome labeled CD3 (BD, 345766), CD4 (BD, 345809), CD8 (BD, 345772), CD314 (BD, 558071), CD45RA (BD, 550855), CD62L (BD, 555544), CD279 (eBioscience, 12-2799-42), CD19 (BD, 345791, CD223 (eBioscience, 25-2239-41), MICA/B (R&D Systems, FAB13001G-100), MICB (R&D Systems, FAB1599G), ULBP1 (R&D Systems, FAB1380C), ULBP2/5/6 (R&D Systems, FAB1298A), ULBP3 (R&D Systems, FAB1517P), ULBP4 (R&D Systems: FAB6285A), and corresponding isotypes according to standard protocols. .. Briefly, cells were harvested and resuspended in a buffer containing DPBS (Life Technologies, A1285801) supplemented with 5% human serum albumin (Octapharma, 68209-633-02) and 0.01% NaN3 (Sigma, S2002).

    Western Blot:

    Article Title: Nano-Engineered Disruption of Heat shock protein 90 (Hsp90) Targets Drug-Induced Resistance and Relieves Natural Killer Cell Suppression in Breast Cancer
    Article Snippet: .. Immunoblotting Protein samples were resolved by SDS-PAGE and transferred to PVDF membranes prior to incubation at 4°C with indicated primary antibodies, mTOR and pMTOR Ser2448 , pAKT Thr308 and AKT, Phospho-p44/42 MAPK (Erk1/2) Thr202/Tyr204 , p44/42 MAPK (Erk1/2) pPRAS40 Thr246 , pSTAT3 Tyr705 , STAT3, PRAS40 and β-Actin antibodies were purchased from Cell Signaling Technology, pHck Tyr410 (Thermo Fisher Scientific) MICA/B (R&D Systems, Minneapolis, MN) and HSF-1 and HSF-1 Ser326 (abcam). ..



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    <t>MICA/B</t> <t>and</t> ULBP2 show different localizations according to the tumor stage. (A) Representative immunohistochemical staining of MICA/B and ULBP2 is shown in adjacent normal tissues (ANT; n = 20), triple-negative breast cancer (TNBC; n = 33) and Luminal A type breast cancer (n = 47). Scale bar = 10 μm. (B,C) Mean score of the distribution of NKG2SLs in the cytosol/membrane fraction compared to the nuclei/perinuclear fraction in either TNBC or Luminal A breast cancer. (D) Representative immunohistochemical staining of MICA/B and ULBP2 is shown in bone metastasis (BoMet; n = 10) and paired bone metastatic ductal carcinoma (bmDC; n = 7). Scale bar = 10 μm. (E,F) Mean score of the distribution of NKG2SLs in the cytosol/membrane fraction compared to the nuclei/perinuclear fraction. Mean of scores derived from the semi-quantitative analysis for MICA/B and ULBP2. Each bar represents the mean ± SD of the specimens, *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.
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    Image Search Results


    MICA/B and ULBP2 show different localizations according to the tumor stage. (A) Representative immunohistochemical staining of MICA/B and ULBP2 is shown in adjacent normal tissues (ANT; n = 20), triple-negative breast cancer (TNBC; n = 33) and Luminal A type breast cancer (n = 47). Scale bar = 10 μm. (B,C) Mean score of the distribution of NKG2SLs in the cytosol/membrane fraction compared to the nuclei/perinuclear fraction in either TNBC or Luminal A breast cancer. (D) Representative immunohistochemical staining of MICA/B and ULBP2 is shown in bone metastasis (BoMet; n = 10) and paired bone metastatic ductal carcinoma (bmDC; n = 7). Scale bar = 10 μm. (E,F) Mean score of the distribution of NKG2SLs in the cytosol/membrane fraction compared to the nuclei/perinuclear fraction. Mean of scores derived from the semi-quantitative analysis for MICA/B and ULBP2. Each bar represents the mean ± SD of the specimens, *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.

    Journal: Frontiers in Cell and Developmental Biology

    Article Title: Breast cancer bone metastasis and bone metastatic cells retain NKG2DLs intracellularly: could this be a strategy to evade immune recognition?

    doi: 10.3389/fcell.2026.1717607

    Figure Lengend Snippet: MICA/B and ULBP2 show different localizations according to the tumor stage. (A) Representative immunohistochemical staining of MICA/B and ULBP2 is shown in adjacent normal tissues (ANT; n = 20), triple-negative breast cancer (TNBC; n = 33) and Luminal A type breast cancer (n = 47). Scale bar = 10 μm. (B,C) Mean score of the distribution of NKG2SLs in the cytosol/membrane fraction compared to the nuclei/perinuclear fraction in either TNBC or Luminal A breast cancer. (D) Representative immunohistochemical staining of MICA/B and ULBP2 is shown in bone metastasis (BoMet; n = 10) and paired bone metastatic ductal carcinoma (bmDC; n = 7). Scale bar = 10 μm. (E,F) Mean score of the distribution of NKG2SLs in the cytosol/membrane fraction compared to the nuclei/perinuclear fraction. Mean of scores derived from the semi-quantitative analysis for MICA/B and ULBP2. Each bar represents the mean ± SD of the specimens, *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.

    Article Snippet: The following primary antibodies were used: MICA/B (1:50, sc-137242, Santa Cruz Biotechnology), ULBP2 (1:100, MA5-29636, Invitrogen), β-tubulin (1:500, T4026, Merck Millipore), β-tubulin (1:100, 2128, Cell Signaling Technology), calnexin (1:100, MA3-027, Invitrogen), calnexin (1:50, 2679, Cell Signaling Technology), GM130 (1:100, ab169276, AbCam), GM130 (1:3200, 1248, Cell Signaling Technology).

    Techniques: Immunohistochemical staining, Staining, Membrane, Derivative Assay

    MICA/B and ULBP2 are differentially expressed in BoMet and BC cell lines with different invasive potential. (A) mRNA expression levels of the NKG2DLs in different BC cell lines (1833, MDA-231, and MCF7) and non-tumorigenic breast cell line (MCF10A). Results are expressed as 2 −ΔΔCt compared to MCF7. ONE-way ANOVA: *p < 0.05; **p < 0.01; n = 3. (B) Western blotting analysis and bands intensity quantification of MICA/B and ULBP2 in BC cell lysates. The expression of MICA/B and ULBP2 was normalized on β-actin. Statistical analysis was performed using Kruskall-Wallis test for non-parametric distribution: *p < 0.05, n = 3.

    Journal: Frontiers in Cell and Developmental Biology

    Article Title: Breast cancer bone metastasis and bone metastatic cells retain NKG2DLs intracellularly: could this be a strategy to evade immune recognition?

    doi: 10.3389/fcell.2026.1717607

    Figure Lengend Snippet: MICA/B and ULBP2 are differentially expressed in BoMet and BC cell lines with different invasive potential. (A) mRNA expression levels of the NKG2DLs in different BC cell lines (1833, MDA-231, and MCF7) and non-tumorigenic breast cell line (MCF10A). Results are expressed as 2 −ΔΔCt compared to MCF7. ONE-way ANOVA: *p < 0.05; **p < 0.01; n = 3. (B) Western blotting analysis and bands intensity quantification of MICA/B and ULBP2 in BC cell lysates. The expression of MICA/B and ULBP2 was normalized on β-actin. Statistical analysis was performed using Kruskall-Wallis test for non-parametric distribution: *p < 0.05, n = 3.

    Article Snippet: The following primary antibodies were used: MICA/B (1:50, sc-137242, Santa Cruz Biotechnology), ULBP2 (1:100, MA5-29636, Invitrogen), β-tubulin (1:500, T4026, Merck Millipore), β-tubulin (1:100, 2128, Cell Signaling Technology), calnexin (1:100, MA3-027, Invitrogen), calnexin (1:50, 2679, Cell Signaling Technology), GM130 (1:100, ab169276, AbCam), GM130 (1:3200, 1248, Cell Signaling Technology).

    Techniques: Expressing, Western Blot

    MICA/B and ULBP2 are expressed both on the surface and intracellularly in BoMet and BC cell lines with different invasive potential. (A) Surface staining of MICA/B and ULBP2 (green) in non-permeabilized breast cancer cells. Nuclei are stained with DAPI (blue). (B) Immunofluorescence detection of intracellular MICA/B and ULBP2 (green) in breast cancer cell lines. β-tubulin is marked in red; nuclei are stained with DAPI (blue). (C) Flow cytometry analysis of cell surface and intracellular MICA/B and ULBP2 expression. For surface staining cells were fixed with PFA and incubated with MICA/B-APC or ULBP2-PE primary antibody. For intracellular staining cells were fixed, permeabilized with 0.2% Tween-20, and incubated with primary antibody. Results are expressed as median fluorescence intensity (MFI) of each target after its isotype control subtraction. Intra-group statistical analysis was performed using ONE-way ANOVA with Tuckey’s multiple comparisons test: *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001; n = 3.

    Journal: Frontiers in Cell and Developmental Biology

    Article Title: Breast cancer bone metastasis and bone metastatic cells retain NKG2DLs intracellularly: could this be a strategy to evade immune recognition?

    doi: 10.3389/fcell.2026.1717607

    Figure Lengend Snippet: MICA/B and ULBP2 are expressed both on the surface and intracellularly in BoMet and BC cell lines with different invasive potential. (A) Surface staining of MICA/B and ULBP2 (green) in non-permeabilized breast cancer cells. Nuclei are stained with DAPI (blue). (B) Immunofluorescence detection of intracellular MICA/B and ULBP2 (green) in breast cancer cell lines. β-tubulin is marked in red; nuclei are stained with DAPI (blue). (C) Flow cytometry analysis of cell surface and intracellular MICA/B and ULBP2 expression. For surface staining cells were fixed with PFA and incubated with MICA/B-APC or ULBP2-PE primary antibody. For intracellular staining cells were fixed, permeabilized with 0.2% Tween-20, and incubated with primary antibody. Results are expressed as median fluorescence intensity (MFI) of each target after its isotype control subtraction. Intra-group statistical analysis was performed using ONE-way ANOVA with Tuckey’s multiple comparisons test: *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001; n = 3.

    Article Snippet: The following primary antibodies were used: MICA/B (1:50, sc-137242, Santa Cruz Biotechnology), ULBP2 (1:100, MA5-29636, Invitrogen), β-tubulin (1:500, T4026, Merck Millipore), β-tubulin (1:100, 2128, Cell Signaling Technology), calnexin (1:100, MA3-027, Invitrogen), calnexin (1:50, 2679, Cell Signaling Technology), GM130 (1:100, ab169276, AbCam), GM130 (1:3200, 1248, Cell Signaling Technology).

    Techniques: Staining, Immunofluorescence, Flow Cytometry, Expressing, Incubation, Fluorescence, Control

    Intracellular MICA/B and ULBP2 co-localize with Golgi apparatus and show different mechanism of intracellular trafficking. (A) Subcellular localization of MICA/B and ULBP2 (green) in BC cell lines, and co-localization with calnexin (endoplasmic reticulum, ER) and GM130 (Golgi) (red). Nuclei are counterstained with DAPI (blue). White arrows indicate co-localization (yellow/orange staining). (B) Calnexin or GM130 co-localization with MICA/B or ULBP2 was quantified by ImageJ plug-in JACOP. Experiment was performed twice and analyzed in at least 6 fields for each condition. Statistical analysis was performed using Kruskall-Wallis for non-parametric distribution with Dunn’s multiple comparisons within organelles: *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. (C) Western blotting analysis of NKG2DLs and GM130 in subcellular fractions: cytosol (C), membrane (M), nucleus (N) and cytoskeleton (CS) extracts. TATA-binding protein (TBP), β-Actin and Na + /K + -ATPase were used as housekeeping for each fraction.

    Journal: Frontiers in Cell and Developmental Biology

    Article Title: Breast cancer bone metastasis and bone metastatic cells retain NKG2DLs intracellularly: could this be a strategy to evade immune recognition?

    doi: 10.3389/fcell.2026.1717607

    Figure Lengend Snippet: Intracellular MICA/B and ULBP2 co-localize with Golgi apparatus and show different mechanism of intracellular trafficking. (A) Subcellular localization of MICA/B and ULBP2 (green) in BC cell lines, and co-localization with calnexin (endoplasmic reticulum, ER) and GM130 (Golgi) (red). Nuclei are counterstained with DAPI (blue). White arrows indicate co-localization (yellow/orange staining). (B) Calnexin or GM130 co-localization with MICA/B or ULBP2 was quantified by ImageJ plug-in JACOP. Experiment was performed twice and analyzed in at least 6 fields for each condition. Statistical analysis was performed using Kruskall-Wallis for non-parametric distribution with Dunn’s multiple comparisons within organelles: *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. (C) Western blotting analysis of NKG2DLs and GM130 in subcellular fractions: cytosol (C), membrane (M), nucleus (N) and cytoskeleton (CS) extracts. TATA-binding protein (TBP), β-Actin and Na + /K + -ATPase were used as housekeeping for each fraction.

    Article Snippet: The following primary antibodies were used: MICA/B (1:50, sc-137242, Santa Cruz Biotechnology), ULBP2 (1:100, MA5-29636, Invitrogen), β-tubulin (1:500, T4026, Merck Millipore), β-tubulin (1:100, 2128, Cell Signaling Technology), calnexin (1:100, MA3-027, Invitrogen), calnexin (1:50, 2679, Cell Signaling Technology), GM130 (1:100, ab169276, AbCam), GM130 (1:3200, 1248, Cell Signaling Technology).

    Techniques: Staining, Western Blot, Membrane, Binding Assay

    MICA/B and ULBP2 are resistant to EndoH digestion but sensitive to PNGase F digestion. (A) Western blotting analysis of MICA/B and ULBP2 in cell lysates from BC cell lines, untreated or treated with 1500 U EndoH for 4 h. The bars represent the mean percentage of digested (□) and undigested (■) MICA/B and ULBP2 normalized on β-actin. Statistical analysis was performed using two-way ANOVA followed by Sidak’s multiple comparison test within cell line: *p < 0.05; **p < 0.001; ***p < 0.0001; n = 3. (B) Western blotting analysis of MICA/B and ULBP2 in cell lysates from BC cell lines, untreated or treated with 20 U PNGase F for 2 h. The bars represent the mean percentage of digested (□) and undigested (■) MICA/B and ULBP2 normalized on β-actin. Statistical analysis was performed using two-way ANOVA followed by Sidak’s multiple comparison test within cell line: **p < 0.001; ***p < 0.0001; n = 3.

    Journal: Frontiers in Cell and Developmental Biology

    Article Title: Breast cancer bone metastasis and bone metastatic cells retain NKG2DLs intracellularly: could this be a strategy to evade immune recognition?

    doi: 10.3389/fcell.2026.1717607

    Figure Lengend Snippet: MICA/B and ULBP2 are resistant to EndoH digestion but sensitive to PNGase F digestion. (A) Western blotting analysis of MICA/B and ULBP2 in cell lysates from BC cell lines, untreated or treated with 1500 U EndoH for 4 h. The bars represent the mean percentage of digested (□) and undigested (■) MICA/B and ULBP2 normalized on β-actin. Statistical analysis was performed using two-way ANOVA followed by Sidak’s multiple comparison test within cell line: *p < 0.05; **p < 0.001; ***p < 0.0001; n = 3. (B) Western blotting analysis of MICA/B and ULBP2 in cell lysates from BC cell lines, untreated or treated with 20 U PNGase F for 2 h. The bars represent the mean percentage of digested (□) and undigested (■) MICA/B and ULBP2 normalized on β-actin. Statistical analysis was performed using two-way ANOVA followed by Sidak’s multiple comparison test within cell line: **p < 0.001; ***p < 0.0001; n = 3.

    Article Snippet: The following primary antibodies were used: MICA/B (1:50, sc-137242, Santa Cruz Biotechnology), ULBP2 (1:100, MA5-29636, Invitrogen), β-tubulin (1:500, T4026, Merck Millipore), β-tubulin (1:100, 2128, Cell Signaling Technology), calnexin (1:100, MA3-027, Invitrogen), calnexin (1:50, 2679, Cell Signaling Technology), GM130 (1:100, ab169276, AbCam), GM130 (1:3200, 1248, Cell Signaling Technology).

    Techniques: Western Blot, Comparison

    NKG2DLs glycosylation and surface localization are differentially affected by tunicamycin in metastatic and non-metastatic BC cells and depend on intact Golgi for intracellular retention. (A) Western blotting analysis and bands intensity quantification of MICA/B and ULBP2 in BC cells after treatment with 1500 U tunicamycin for 24 h. The bars represent the mean percentage of digested (□) and undigested (■) MICA/B and ULBP2 normalized on β-actin. Statistical analysis was performed using two-way ANOVA followed by Sidak’s multiple comparison test within cell line: ***p < 0.0001; or using two-way ANOVA followed by Tukey’s multiple comparison test between cell lines: °p < 0.05; °°p < 0.01; °°°p < 0.001; °°°°p < 0.0001; n = 3. (B) Flow cytometry analysis of cell surface MICA/B and ULBP2 expression after treatment with 1500 U tunicamycin for 24 h. For surface staining cells were incubated with MICA/B-APC or ULBP2-APC primary antibody. Representative histograms overlay are shown for each cell line. Results are expressed as relative median fluorescence intensity (MFI) of each target to its untreated control (NT). Statistical analysis was performed using ONE-way ANOVA with Sidak’s multiple comparisons test within cell lines: *p < 0.05; ***p < 0.001; n = 3. (C) Subcellular localization of MICA/B and ULBP2 (green) in BC cell lines, and co-localization with GM130 (Golgi) (red) after treatment with 2 µM monensin for 3 h. Nuclei are counterstained with DAPI (blue).

    Journal: Frontiers in Cell and Developmental Biology

    Article Title: Breast cancer bone metastasis and bone metastatic cells retain NKG2DLs intracellularly: could this be a strategy to evade immune recognition?

    doi: 10.3389/fcell.2026.1717607

    Figure Lengend Snippet: NKG2DLs glycosylation and surface localization are differentially affected by tunicamycin in metastatic and non-metastatic BC cells and depend on intact Golgi for intracellular retention. (A) Western blotting analysis and bands intensity quantification of MICA/B and ULBP2 in BC cells after treatment with 1500 U tunicamycin for 24 h. The bars represent the mean percentage of digested (□) and undigested (■) MICA/B and ULBP2 normalized on β-actin. Statistical analysis was performed using two-way ANOVA followed by Sidak’s multiple comparison test within cell line: ***p < 0.0001; or using two-way ANOVA followed by Tukey’s multiple comparison test between cell lines: °p < 0.05; °°p < 0.01; °°°p < 0.001; °°°°p < 0.0001; n = 3. (B) Flow cytometry analysis of cell surface MICA/B and ULBP2 expression after treatment with 1500 U tunicamycin for 24 h. For surface staining cells were incubated with MICA/B-APC or ULBP2-APC primary antibody. Representative histograms overlay are shown for each cell line. Results are expressed as relative median fluorescence intensity (MFI) of each target to its untreated control (NT). Statistical analysis was performed using ONE-way ANOVA with Sidak’s multiple comparisons test within cell lines: *p < 0.05; ***p < 0.001; n = 3. (C) Subcellular localization of MICA/B and ULBP2 (green) in BC cell lines, and co-localization with GM130 (Golgi) (red) after treatment with 2 µM monensin for 3 h. Nuclei are counterstained with DAPI (blue).

    Article Snippet: The following primary antibodies were used: MICA/B (1:50, sc-137242, Santa Cruz Biotechnology), ULBP2 (1:100, MA5-29636, Invitrogen), β-tubulin (1:500, T4026, Merck Millipore), β-tubulin (1:100, 2128, Cell Signaling Technology), calnexin (1:100, MA3-027, Invitrogen), calnexin (1:50, 2679, Cell Signaling Technology), GM130 (1:100, ab169276, AbCam), GM130 (1:3200, 1248, Cell Signaling Technology).

    Techniques: Glycoproteomics, Western Blot, Comparison, Flow Cytometry, Expressing, Staining, Incubation, Fluorescence, Control

    Schematic representation of MICA/B and ULBP2 maturation and trafficking through Golgi apparatus in metastatic and non-metastatic BC cell lines. In metastatic cell lines (1833 and MDA-231), following protein synthesis, NKG2DLs acquire complex N-glycans and traffic from the endoplasmic reticulum (ER) to the Golgi apparatus. When N-glycosylation is impaired (e.g., by tunicamycin treatment), NKG2DLs fail to reach the cell surface. In contrast, in the non-metastatic MCF7 cell line, which is largely resistant to tunicamycin, immature NKG2DLs are still transported through the Golgi apparatus and delivered to the plasma membrane, resulting in increased surface expression irrespective of correct glycosylation. Nevertheless, regardless of glycosylation status, delivery to the plasma membrane requires an intact Golgi architecture, as demonstrated by the loss of co-localization with a Golgi marker following monensin treatment. Created in BioRender. Lombardi, G. (2025) https://BioRender.com/jnrm8ui .

    Journal: Frontiers in Cell and Developmental Biology

    Article Title: Breast cancer bone metastasis and bone metastatic cells retain NKG2DLs intracellularly: could this be a strategy to evade immune recognition?

    doi: 10.3389/fcell.2026.1717607

    Figure Lengend Snippet: Schematic representation of MICA/B and ULBP2 maturation and trafficking through Golgi apparatus in metastatic and non-metastatic BC cell lines. In metastatic cell lines (1833 and MDA-231), following protein synthesis, NKG2DLs acquire complex N-glycans and traffic from the endoplasmic reticulum (ER) to the Golgi apparatus. When N-glycosylation is impaired (e.g., by tunicamycin treatment), NKG2DLs fail to reach the cell surface. In contrast, in the non-metastatic MCF7 cell line, which is largely resistant to tunicamycin, immature NKG2DLs are still transported through the Golgi apparatus and delivered to the plasma membrane, resulting in increased surface expression irrespective of correct glycosylation. Nevertheless, regardless of glycosylation status, delivery to the plasma membrane requires an intact Golgi architecture, as demonstrated by the loss of co-localization with a Golgi marker following monensin treatment. Created in BioRender. Lombardi, G. (2025) https://BioRender.com/jnrm8ui .

    Article Snippet: The following primary antibodies were used: MICA/B (1:50, sc-137242, Santa Cruz Biotechnology), ULBP2 (1:100, MA5-29636, Invitrogen), β-tubulin (1:500, T4026, Merck Millipore), β-tubulin (1:100, 2128, Cell Signaling Technology), calnexin (1:100, MA3-027, Invitrogen), calnexin (1:50, 2679, Cell Signaling Technology), GM130 (1:100, ab169276, AbCam), GM130 (1:3200, 1248, Cell Signaling Technology).

    Techniques: Glycoproteomics, Clinical Proteomics, Membrane, Expressing, Marker