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R&D Systems cd44v6 fitc antibody
Cd44v6 Fitc Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 443 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/cd44v6+fitc+antibody/FITC+Antibody/us10206875-307-129-134
Average 95 stars, based on 443 article reviews

cd44v6 fitc antibody - by Bioz Stars, 2026-09

95/100 stars

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Proliferation Assay:

Article Title: Nanoemulsion Formulation of a Novel Taxoid DHA-SBT-1214 Inhibits Prostate Cancer Stem Cell-Induced Tumor Growth
Article Snippet: 2.1 Materials New-generation taxoid, DHA-SBT-1214 was synthesized by ChemMaster International, Inc. (Stony Brook, NY). .. Extra pure omega-3 rich fish oil was purchased from Jedwards International (Quincy, MA), Lipoid E80 from Lipoid GMBH (Ludwigshafen, Germany), DSPE PEG2000 from Avanti Polar Lipids, Inc. (Alabaster, AL), Tween 80 from Sigma Chemicals, Inc. (St. Louis, MO), CellTiter 96 AQ ueous one solution cell proliferation assay kit (G3580), from Promega (Madison, WI), Mesenchymal stem cell growth media (MSCGM) from Lonza (Portsmouth, NH), LAL chromogenic endotoxin quantitation kit from Thermo Scientific (Rockford, IL), Microscope slides single depression concave from Amscope (Irvine, CA), Collagenases type II and type IV from Sigma-Aldrich, Rhodamine 123 from Sigma Aldrich (St. Louis, MO), Anti-human CD133/2-APC antibody (clone 293C3) from Miltenyi Biotec, CA, USA; CD44-FITC antibody (clone F10-44-2) or CD44-PE antibody (clone F10-44-2) from; Invitrogen/Biosources, USA; CD166-PE antibody (clone 105902) and CD44v6-FITC antibody (clone 2F10) from; R&D Systems, MN, USA, EpCAM-FITC antibody from Biosource, CA, USA, Pan-Keratin (C11) antibody-Alexa Fluor ® 488 from Cell Signaling and all the isotype controls antibodies were purchased from Chemicon. .. Penicillin, streptomycin and Trypsin-EDTA were obtained from Invitrogen (Grand Island, NY, USA).

Article Title: Nanoemulsion compositions of taxoid drugs, and methods for the use thereof to target cancer cells and cancer stem cells
Article Snippet: Materials and Methods New-generation taxoid, DHA-SBT-1214 was synthesized by Dr. Ojima's laboratory at Stony Brook University, (Stony Brook, N.Y.) or ChemMaster International, Inc. (Stony Brook, N.Y.). .. Extra pure omega-3 rich fish oil was purchased from Jedwards International (Quincy, Mass.), Lipoid E80 from Lipoid GMBH (Ludwigshafen, Germany), DSPE PEG2000 from Avanti Polar Lipids, Inc. (Alabaster, Ala.), Tween 80 from Sigma Chemicals, Inc. (St. Louis, Mo.), CellTiter 96 AQueous one solution cell proliferation assay kit (G3580), from Promega (Madison, Wis.), Mesenchymal stem cell growth media (MSCGM) from Lonza (Portsmouth, N.H.), LAL chromogenic endotoxin quantitation kit from Thermo Scientific (Rockford, Ill.), Microscope slides single depression concave from Amscope (Irvine, Calif.), Collagenases type II and type IV from Sigma-Aldrich, Rhodamine 123 from Sigma Aldrich (St. Louis, Mo.), Anti-human CD133/2-APC antibody (clone 293C3) from Miltenyi Biotec, CA, USA; CD166-PE antibody (clone 105902) from R&D Systems, MN, USA; CD44-FITC antibody (clone F10-44-2) from or CD44-PE antibody (clone F10-44-2) from; Invitrogen/Biosources, USA; CD44v6-FITC antibody (clone 2F10) from; R&D Systems, USA, EpCAM-FITC antibody from Biosource, CA, USA, Pan-Keratin (C11) antibody—ALEXAFLUOR® 488 from Cell Signaling and all the isotype controls antibodies were purchased from Chemicon. .. Penicillin, streptomycin and TrypLE were obtained from Invitrogen (Grand Island, N.Y., USA).

Quantitation Assay:

Microscopy:

Image Search Results

Phenotypic and functional characterization of CD44v6 chimeric antigen receptor (CAR)-T cells. (A) Memory/effector status of CAR-T cells before and after puromycin selection. Flow cytometry was performed using antibodies against the Fc gamma region, which binds to the hinge domain of the CAR expressed on both CD4 + and CD8 + T cells. Representative scatter plots are shown for donor 1-derived CAR-T cells. (B) Memory/effector subset analysis of CAR-T cells following co-culture with or without the indicated cell lines, followed by incubation in cytokine-depleted medium for 3 d Cells were stained with CD62L and CD45RO and gated on CD45 + CD3 + CAR-T cells for flow cytometric analysis. Representative data from donor 1-derived CAR-T cells are shown. (C) Immunofluorescence analysis of antigen-dependent binding of CAR-T cells. Donor 1-derived CAR-T cells were co-cultured for 3 h with Panc-1 (CD44v6-positive) or NIH/3T3 (CD44v6-negative) cells, then stained with anti-CD3 antibody. Fluorescent labeling: CD3 (green), CD44v6 (red), and nuclei (DAPI, blue). (D) Antigen-dependent activation and degranulation of CAR-T cells. Cells were co-cultured for 24 h with or without target cell lines in cytokine-depleted medium in the presence of CD107a antibody and the Golgi transport inhibitor Monensin. Activation and degranulation were assessed using CD69 and CD8a staining. Representative flow cytometry plots are shown. (E) Cytokine secretion by CAR-T cells following co-culture with the indicated cell lines. Levels of TNF-α and IFN-γ were quantified by ELISA after 48 h at an E:T ratio of 1:1. Data are presented as mean ± standard deviation (SD) from three independent experiments using CAR-T cells derived from donors 1, 2, and 3. (F) Cytolytic activity of control T and donor 1-derived CAR-T cells against various cancer cell lines. Cells were co-cultured for 3 d and their viability was assessed using crystal violet staining. Absorbance values were normalized to untreated wells to calculate relative cancer cell survival. Data are presented as mean ± SD of three independent experiments performed in triplicate. ns, not significant; ** P < 0.01; *** P < 0.001 (two-way ANOVA).

Journal: Frontiers in Immunology

Article Title: Relaxin-2-secreting CAR-T cells exhibit enhanced efficacy in stromal-rich xenograft tumors

doi: 10.3389/fimmu.2025.1506204

Figure Lengend Snippet: Phenotypic and functional characterization of CD44v6 chimeric antigen receptor (CAR)-T cells. (A) Memory/effector status of CAR-T cells before and after puromycin selection. Flow cytometry was performed using antibodies against the Fc gamma region, which binds to the hinge domain of the CAR expressed on both CD4 + and CD8 + T cells. Representative scatter plots are shown for donor 1-derived CAR-T cells. (B) Memory/effector subset analysis of CAR-T cells following co-culture with or without the indicated cell lines, followed by incubation in cytokine-depleted medium for 3 d Cells were stained with CD62L and CD45RO and gated on CD45 + CD3 + CAR-T cells for flow cytometric analysis. Representative data from donor 1-derived CAR-T cells are shown. (C) Immunofluorescence analysis of antigen-dependent binding of CAR-T cells. Donor 1-derived CAR-T cells were co-cultured for 3 h with Panc-1 (CD44v6-positive) or NIH/3T3 (CD44v6-negative) cells, then stained with anti-CD3 antibody. Fluorescent labeling: CD3 (green), CD44v6 (red), and nuclei (DAPI, blue). (D) Antigen-dependent activation and degranulation of CAR-T cells. Cells were co-cultured for 24 h with or without target cell lines in cytokine-depleted medium in the presence of CD107a antibody and the Golgi transport inhibitor Monensin. Activation and degranulation were assessed using CD69 and CD8a staining. Representative flow cytometry plots are shown. (E) Cytokine secretion by CAR-T cells following co-culture with the indicated cell lines. Levels of TNF-α and IFN-γ were quantified by ELISA after 48 h at an E:T ratio of 1:1. Data are presented as mean ± standard deviation (SD) from three independent experiments using CAR-T cells derived from donors 1, 2, and 3. (F) Cytolytic activity of control T and donor 1-derived CAR-T cells against various cancer cell lines. Cells were co-cultured for 3 d and their viability was assessed using crystal violet staining. Absorbance values were normalized to untreated wells to calculate relative cancer cell survival. Data are presented as mean ± SD of three independent experiments performed in triplicate. ns, not significant; ** P < 0.01; *** P < 0.001 (two-way ANOVA).

Article Snippet: The expression of CD44v6 in various cancer cell lines was assessed using a mouse anti-human FITC-conjugated CD44v6 antibody (clone VFF-18, eBioscience).

Techniques: Functional Assay, Selection, Flow Cytometry, Derivative Assay, Co-Culture Assay, Incubation, Staining, Immunofluorescence, Binding Assay, Cell Culture, Labeling, Activation Assay, Enzyme-linked Immunosorbent Assay, Standard Deviation, Activity Assay, Control

Development of CD44v6 CAR-T cells and engineering for RLN2 secretion. (A) Schematic illustration of plasmid constructs used for CAR-T cell generation, including CD44v6 CAR and co-expression of either Luc2 or human RLN2 via transposon-based vectors. (B) RLN2 protein levels secreted by conventional and RLN2-secreting CAR-T cells quantified using ELISA. Data are presented as mean ± SD from three independent experiments performed in triplicate using CAR-T cells derived from donors 1, 2, and 3. (C) Western blot analysis of the RLN2 receptor LGR7/RXFP1 in multiple cancer cell lines. GAPDH was used as a loading control. (D) Expression of MMPs in cancer cell lines cultured for 48 h in serum-free medium (SF), 1% fetal bovine serum (FBS)-containing medium (1% CM), supernatant from CAR-T cells (CAR-T sup), or supernatant from RLN2-secreting CAR-T cells (RLN2 CAR-T sup), collected after 24 h in SF. RT-qPCR was conducted in three independent runs using CAR-T cells from donors 1, 2, and 3, in triplicate (mean ± SD). * P < 0.05; ** P < 0.01; *** P < 0.001; ns, not significant (one-way ANOVA followed by Bonferroni’s post hoc test and unpaired t -test for comparisons between CAR-T and RLN2-secreting CAR-T groups). (E) Gelatin zymography of conditioned media from SU86.86 cells cultured in SF, complete medium with 10% FBS (CM), or undiluted (×1) or 5-fold diluted (×1/5) supernatants from CAR-T or RLN2 CAR-T cells. (F) Western blot analysis of LGR7/RXFP1 expression in conventional and RLN2-secreting CAR-T cells. MMP expression in imhPSCs treated with the indicated supernatants (evaluated as described in D ). (G) Western blot analysis of α-SMA and collagen type I alpha 1 (COL1A1) in imhPSCs cultured for 48 h in SF, SF + TGF-β (10 ng/mL), CAR-T sup ± TGF-β, or RLN2 CAR-T sup ± TGF-β. GAPDH was used as a loading control. Supernatants were derived from donor 1-generated CAR-T cells. (H) MMP expression in CAR-T cells. Conventional and RLN2-secreting CAR-T cells were cultured in RPMI-1640 with 10% FBS under resting conditions or with stimulation (anti-CD3/CD28 antibodies + IL-15 and IL-21) for 3 d RT-qPCR was performed using cells from donors 1, 2, and 3 (mean ± SD; triplicate experiments). * P < 0.05; *** P < 0.001; ns, not significant (comparisons between CAR-T and RLN2 CAR-T cells analyzed using unpaired t -test).

Journal: Frontiers in Immunology

Article Title: Relaxin-2-secreting CAR-T cells exhibit enhanced efficacy in stromal-rich xenograft tumors

doi: 10.3389/fimmu.2025.1506204

Figure Lengend Snippet: Development of CD44v6 CAR-T cells and engineering for RLN2 secretion. (A) Schematic illustration of plasmid constructs used for CAR-T cell generation, including CD44v6 CAR and co-expression of either Luc2 or human RLN2 via transposon-based vectors. (B) RLN2 protein levels secreted by conventional and RLN2-secreting CAR-T cells quantified using ELISA. Data are presented as mean ± SD from three independent experiments performed in triplicate using CAR-T cells derived from donors 1, 2, and 3. (C) Western blot analysis of the RLN2 receptor LGR7/RXFP1 in multiple cancer cell lines. GAPDH was used as a loading control. (D) Expression of MMPs in cancer cell lines cultured for 48 h in serum-free medium (SF), 1% fetal bovine serum (FBS)-containing medium (1% CM), supernatant from CAR-T cells (CAR-T sup), or supernatant from RLN2-secreting CAR-T cells (RLN2 CAR-T sup), collected after 24 h in SF. RT-qPCR was conducted in three independent runs using CAR-T cells from donors 1, 2, and 3, in triplicate (mean ± SD). * P < 0.05; ** P < 0.01; *** P < 0.001; ns, not significant (one-way ANOVA followed by Bonferroni’s post hoc test and unpaired t -test for comparisons between CAR-T and RLN2-secreting CAR-T groups). (E) Gelatin zymography of conditioned media from SU86.86 cells cultured in SF, complete medium with 10% FBS (CM), or undiluted (×1) or 5-fold diluted (×1/5) supernatants from CAR-T or RLN2 CAR-T cells. (F) Western blot analysis of LGR7/RXFP1 expression in conventional and RLN2-secreting CAR-T cells. MMP expression in imhPSCs treated with the indicated supernatants (evaluated as described in D ). (G) Western blot analysis of α-SMA and collagen type I alpha 1 (COL1A1) in imhPSCs cultured for 48 h in SF, SF + TGF-β (10 ng/mL), CAR-T sup ± TGF-β, or RLN2 CAR-T sup ± TGF-β. GAPDH was used as a loading control. Supernatants were derived from donor 1-generated CAR-T cells. (H) MMP expression in CAR-T cells. Conventional and RLN2-secreting CAR-T cells were cultured in RPMI-1640 with 10% FBS under resting conditions or with stimulation (anti-CD3/CD28 antibodies + IL-15 and IL-21) for 3 d RT-qPCR was performed using cells from donors 1, 2, and 3 (mean ± SD; triplicate experiments). * P < 0.05; *** P < 0.001; ns, not significant (comparisons between CAR-T and RLN2 CAR-T cells analyzed using unpaired t -test).

Article Snippet: The expression of CD44v6 in various cancer cell lines was assessed using a mouse anti-human FITC-conjugated CD44v6 antibody (clone VFF-18, eBioscience).

Techniques: Plasmid Preparation, Construct, Expressing, Enzyme-linked Immunosorbent Assay, Derivative Assay, Western Blot, Control, Cell Culture, Quantitative RT-PCR, Zymography, Generated

Histological characterization of subcutaneous xenograft tumor models. (A) Histological analysis of Panc-1, AsPC-1-CD44v6, and SU86.86 xenograft tumors. Tumor sections were analyzed for CD44v6 expression and stromal architecture. Formalin-fixed paraffin-embedded samples were subjected to H&E staining for tissue morphology, Picro-Sirius Red staining for collagen types I and III, and immunohistochemistry for CD44v6 and collagen type IV analysis. Scale bar = 200 µm. (B) Immunofluorescence staining of vascular and stromal markers in Panc-1 and SU86.86 subcutaneous xenograft tumors. Endothelial cells were labeled with anti-CD31 (green), myofibroblasts and pericytes with anti-α-SMA (red), and nuclei with DAPI (blue). Scale bar = 100 µm.

Journal: Frontiers in Immunology

Article Title: Relaxin-2-secreting CAR-T cells exhibit enhanced efficacy in stromal-rich xenograft tumors

doi: 10.3389/fimmu.2025.1506204

Figure Lengend Snippet: Histological characterization of subcutaneous xenograft tumor models. (A) Histological analysis of Panc-1, AsPC-1-CD44v6, and SU86.86 xenograft tumors. Tumor sections were analyzed for CD44v6 expression and stromal architecture. Formalin-fixed paraffin-embedded samples were subjected to H&E staining for tissue morphology, Picro-Sirius Red staining for collagen types I and III, and immunohistochemistry for CD44v6 and collagen type IV analysis. Scale bar = 200 µm. (B) Immunofluorescence staining of vascular and stromal markers in Panc-1 and SU86.86 subcutaneous xenograft tumors. Endothelial cells were labeled with anti-CD31 (green), myofibroblasts and pericytes with anti-α-SMA (red), and nuclei with DAPI (blue). Scale bar = 100 µm.

Article Snippet: The expression of CD44v6 in various cancer cell lines was assessed using a mouse anti-human FITC-conjugated CD44v6 antibody (clone VFF-18, eBioscience).

Techniques: Expressing, Formalin-fixed Paraffin-Embedded, Staining, Immunohistochemistry, Immunofluorescence, Labeling

Efficacy of conventional CD44v6 CAR-T cell therapy in xenograft models with either scant or abundant stroma. (A–C) In vitro cytotoxicity (left) and in vivo antitumor activity (right) of donor 1-derived CD44v6 CAR-T cells against Panc-1 (A) , AsPC-1-CD44v6 (B) , and SU86.86 (C) tumor models. For in vitro assays, CAR-T cells were co-cultured with the indicated cancer cell lines for 3 d, and surviving cancer cells were quantified using a crystal violet assay. Absorbance at 590 nm (A 590 ) was measured and normalized to untreated controls. Data are presented as mean ± SD from three independent experiments conducted in triplicate. ns, not significant; *** P < 0.001 (two-way ANOVA). For in vivo studies, tumor-bearing mice received a single intravenous injection of either Ctrl-T ( n = 6) or CAR-T cells ( n = 6). Data are presented as mean ± SD. *** P < 0.001; ns, not significant (two-way ANOVA). (D) In vivo bioluminescence imaging of Luc2-expressing CAR-T cell accumulation in subcutaneous xenograft tumors. A total of 1 × 10 7 Luc2-expressing CAR-T cells were injected intravenously via the tail vein. Bioluminescence was measured using the IVIS imaging system on days 7 and 14 post-injection. (E) Immunofluorescence analysis of CAR-T cell infiltration in AsPC-1-CD44v6 and SU86.86 xenograft tumors 7 d after systemic injection of Luc2-expressing CAR-T cells. Tumor sections were stained for CD3 (green), CD44v6 (red), and CD31 (blue). Scale bar = 100 µm.

Journal: Frontiers in Immunology

Article Title: Relaxin-2-secreting CAR-T cells exhibit enhanced efficacy in stromal-rich xenograft tumors

doi: 10.3389/fimmu.2025.1506204

Figure Lengend Snippet: Efficacy of conventional CD44v6 CAR-T cell therapy in xenograft models with either scant or abundant stroma. (A–C) In vitro cytotoxicity (left) and in vivo antitumor activity (right) of donor 1-derived CD44v6 CAR-T cells against Panc-1 (A) , AsPC-1-CD44v6 (B) , and SU86.86 (C) tumor models. For in vitro assays, CAR-T cells were co-cultured with the indicated cancer cell lines for 3 d, and surviving cancer cells were quantified using a crystal violet assay. Absorbance at 590 nm (A 590 ) was measured and normalized to untreated controls. Data are presented as mean ± SD from three independent experiments conducted in triplicate. ns, not significant; *** P < 0.001 (two-way ANOVA). For in vivo studies, tumor-bearing mice received a single intravenous injection of either Ctrl-T ( n = 6) or CAR-T cells ( n = 6). Data are presented as mean ± SD. *** P < 0.001; ns, not significant (two-way ANOVA). (D) In vivo bioluminescence imaging of Luc2-expressing CAR-T cell accumulation in subcutaneous xenograft tumors. A total of 1 × 10 7 Luc2-expressing CAR-T cells were injected intravenously via the tail vein. Bioluminescence was measured using the IVIS imaging system on days 7 and 14 post-injection. (E) Immunofluorescence analysis of CAR-T cell infiltration in AsPC-1-CD44v6 and SU86.86 xenograft tumors 7 d after systemic injection of Luc2-expressing CAR-T cells. Tumor sections were stained for CD3 (green), CD44v6 (red), and CD31 (blue). Scale bar = 100 µm.

Article Snippet: The expression of CD44v6 in various cancer cell lines was assessed using a mouse anti-human FITC-conjugated CD44v6 antibody (clone VFF-18, eBioscience).

Techniques: In Vitro, In Vivo, Activity Assay, Derivative Assay, Cell Culture, Crystal Violet Assay, Injection, Imaging, Expressing, Immunofluorescence, Staining

Antitumor effects of RLN2-secreting CD44v6 CAR-T cell therapy in stromal-rich SU86.86 xenograft tumors. (A) Tumor growth in SU86.86 subcutaneous xenografts after single intravenous injection of Ctrl-T (1 × 10 7 cells, n = 6), conventional CD44v6 CAR-T (1 × 10 7 cells, n = 6), or RLN2-secreting CD44v6 CAR-T cells (1 × 10 7 cells, n = 6). Tumor size was measured over time, and representative images of resected tumors were captured following euthanasia. Data are presented as mean ± SD. ns, not significant; *** P < 0.001 (two-way ANOVA with Tukey’s multiple comparisons test). (B) Histological analysis of SU86.86 xenograft tumors collected 7 d after treatment with conventional or RLN2-secreting CAR-T cells. Formalin-fixed paraffin-embedded sections were subjected to H&E staining, Picro-Sirius Red staining (collagen types I and III), and immunohistochemistry. Immunofluorescence analysis of tumor-infiltrating CAR-T cells was performed on frozen sections. Tumor sections were stained for CD44v6 (red), CD3 (green), and CD31 (blue). Scale bar = 100 µm. (C) Quantitative analysis of MMP expression in SU86.86 xenograft tumors treated with conventional or RLN2-secreting CAR-T cells and harvested at either 3 or 7 d after single treatment with 1 × 10 7 CAR-T cells. MMP-7 and MMP-9 mRNA levels were assessed using RT-qPCR. CAR-T group ( n = 9, day 3; n = 5, day 7), RLN2 CAR-T group ( n = 7, day 3; n = 7, day 7). Data from two independent experiments are shown as mean ± SD. ns, not significant; * P < 0.05; *** P < 0.001; ns, not significant (unpaired t -test).

Journal: Frontiers in Immunology

Article Title: Relaxin-2-secreting CAR-T cells exhibit enhanced efficacy in stromal-rich xenograft tumors

doi: 10.3389/fimmu.2025.1506204

Figure Lengend Snippet: Antitumor effects of RLN2-secreting CD44v6 CAR-T cell therapy in stromal-rich SU86.86 xenograft tumors. (A) Tumor growth in SU86.86 subcutaneous xenografts after single intravenous injection of Ctrl-T (1 × 10 7 cells, n = 6), conventional CD44v6 CAR-T (1 × 10 7 cells, n = 6), or RLN2-secreting CD44v6 CAR-T cells (1 × 10 7 cells, n = 6). Tumor size was measured over time, and representative images of resected tumors were captured following euthanasia. Data are presented as mean ± SD. ns, not significant; *** P < 0.001 (two-way ANOVA with Tukey’s multiple comparisons test). (B) Histological analysis of SU86.86 xenograft tumors collected 7 d after treatment with conventional or RLN2-secreting CAR-T cells. Formalin-fixed paraffin-embedded sections were subjected to H&E staining, Picro-Sirius Red staining (collagen types I and III), and immunohistochemistry. Immunofluorescence analysis of tumor-infiltrating CAR-T cells was performed on frozen sections. Tumor sections were stained for CD44v6 (red), CD3 (green), and CD31 (blue). Scale bar = 100 µm. (C) Quantitative analysis of MMP expression in SU86.86 xenograft tumors treated with conventional or RLN2-secreting CAR-T cells and harvested at either 3 or 7 d after single treatment with 1 × 10 7 CAR-T cells. MMP-7 and MMP-9 mRNA levels were assessed using RT-qPCR. CAR-T group ( n = 9, day 3; n = 5, day 7), RLN2 CAR-T group ( n = 7, day 3; n = 7, day 7). Data from two independent experiments are shown as mean ± SD. ns, not significant; * P < 0.05; *** P < 0.001; ns, not significant (unpaired t -test).

Article Snippet: The expression of CD44v6 in various cancer cell lines was assessed using a mouse anti-human FITC-conjugated CD44v6 antibody (clone VFF-18, eBioscience).

Techniques: Injection, Formalin-fixed Paraffin-Embedded, Staining, Immunohistochemistry, Immunofluorescence, Expressing, Quantitative RT-PCR

(A) Schematic representation of CD44 exons. (B) Gene expression of CD44v6 in matched cancer and adjacent normal mucosa tissues in 2 independent cohorts (Mann-Whitney U test). (C) CD44v6 gene expression categorized by tumor stages (cohort 1: n = 184, cohort 2: n = 150, 1-way ANOVA). (D) Kaplan-Meier analysis for overall survival (OS) in 2 independent cohorts (log-rank test). (E) Kaplan-Meier analysis of disease-free survival (DFS) in 2 independent cohorts (log-rank test). (F) Forest plots illustrate the Cox hazard proportional analysis based on overall survival and DFS stratified by CD44v6 expression and clinical factors. *P < 0.05, **P < 0.01, ***P < 0.001.

Journal: JCI Insight

Article Title: Cancer stem cell–associated miRNAs serve as prognostic biomarkers in colorectal cancer

doi: 10.1172/jci.insight.125294

Figure Lengend Snippet: (A) Schematic representation of CD44 exons. (B) Gene expression of CD44v6 in matched cancer and adjacent normal mucosa tissues in 2 independent cohorts (Mann-Whitney U test). (C) CD44v6 gene expression categorized by tumor stages (cohort 1: n = 184, cohort 2: n = 150, 1-way ANOVA). (D) Kaplan-Meier analysis for overall survival (OS) in 2 independent cohorts (log-rank test). (E) Kaplan-Meier analysis of disease-free survival (DFS) in 2 independent cohorts (log-rank test). (F) Forest plots illustrate the Cox hazard proportional analysis based on overall survival and DFS stratified by CD44v6 expression and clinical factors. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: SDCSCs derived from HCT116 and HT26 cell lines were stained with CD44v6-FITC monoclonal antibody (catalog MA5-16966; Invitrogen), and propidium iodide was used to eliminate dead cells.

Techniques: Expressing, MANN-WHITNEY

(A) Images of sphere-derived cancer stem cells (SDCSCs) and their respective parental cells. (B) Gene expression of pluripotency markers Oct4 and Nanog in SDCSCs and their parental cell lines (n = 3, Mann-Whitney’s U test). (C) Expression of the stemness-suppressive miRNAs miR-34a and miR-200c in SDCSCs and their parental cell lines (n = 3, Mann-Whitney U test). (D) Cytotoxicity from various doses of 5FU (top 2 panels) and oxaliplatin (bottom 2 panels) was assessed for SDCSCs and their respective parental cells lines, HCT116 and HT29. (E) Relative gene expression of CD44 variants in SDCSCs and their parental cell lines (n = 3, Mann-Whitney U test). (F) CD44v6 protein expression in SDCSCs and their parental cell lines, HCT116 and HT29. (G) Immunofluorescence assay of CD44v6 in SDCSCs from HCT116 cells with FITC staining (left) and DAPI nuclear staining (middle); a merged image is shown on the right. *P < 0.05, **P < 0.01, ***P < 0.001.

Journal: JCI Insight

Article Title: Cancer stem cell–associated miRNAs serve as prognostic biomarkers in colorectal cancer

doi: 10.1172/jci.insight.125294

Figure Lengend Snippet: (A) Images of sphere-derived cancer stem cells (SDCSCs) and their respective parental cells. (B) Gene expression of pluripotency markers Oct4 and Nanog in SDCSCs and their parental cell lines (n = 3, Mann-Whitney’s U test). (C) Expression of the stemness-suppressive miRNAs miR-34a and miR-200c in SDCSCs and their parental cell lines (n = 3, Mann-Whitney U test). (D) Cytotoxicity from various doses of 5FU (top 2 panels) and oxaliplatin (bottom 2 panels) was assessed for SDCSCs and their respective parental cells lines, HCT116 and HT29. (E) Relative gene expression of CD44 variants in SDCSCs and their parental cell lines (n = 3, Mann-Whitney U test). (F) CD44v6 protein expression in SDCSCs and their parental cell lines, HCT116 and HT29. (G) Immunofluorescence assay of CD44v6 in SDCSCs from HCT116 cells with FITC staining (left) and DAPI nuclear staining (middle); a merged image is shown on the right. *P < 0.05, **P < 0.01, ***P < 0.001.

Techniques: Derivative Assay, Expressing, MANN-WHITNEY, Immunofluorescence, Staining

(A) CD44v6 protein expression following transient knockdown (KD) in HT29 and HCT116 cell lines. NC, negative control. (B) Cell proliferation of CD44v6 KD cells and parental cell lines (n = 8 per time point, Mann-Whitney U test). (C) Clonogenicity of HCT116 and HT29 cells with and without KD of CD44v6 (n = 6, Mann-Whitney U test). (D) Spheroid formation capacity of HCT116 and HT29 cells with and without KD of CD44v6 (HCT116: n = 20, HT29: n = 8, Mann-Whitney U test). (E) Cytotoxicity of various doses of 5FU (left) and oxaliplatin (right) was assessed for HCT116 cells with and without KD of CD44v6. *P < 0.05, ***P < 0.001.

Journal: JCI Insight

Article Title: Cancer stem cell–associated miRNAs serve as prognostic biomarkers in colorectal cancer

doi: 10.1172/jci.insight.125294

Figure Lengend Snippet: (A) CD44v6 protein expression following transient knockdown (KD) in HT29 and HCT116 cell lines. NC, negative control. (B) Cell proliferation of CD44v6 KD cells and parental cell lines (n = 8 per time point, Mann-Whitney U test). (C) Clonogenicity of HCT116 and HT29 cells with and without KD of CD44v6 (n = 6, Mann-Whitney U test). (D) Spheroid formation capacity of HCT116 and HT29 cells with and without KD of CD44v6 (HCT116: n = 20, HT29: n = 8, Mann-Whitney U test). (E) Cytotoxicity of various doses of 5FU (left) and oxaliplatin (right) was assessed for HCT116 cells with and without KD of CD44v6. *P < 0.05, ***P < 0.001.

Techniques: Expressing, Negative Control, MANN-WHITNEY

(A) FACS analysis of CD44v6-positive and -negative sphere-derived cancer stem cells (SDCSCs) derived from a HCT116 cell line. (B) Spheroid-forming capacity of HCT116- and HT29-derived CD44v6+ and CD44v6– SDCSCs (n = 16, Mann-Whitney U test). (C) Spheroid-forming ability of HCT116 (left) and HT29 (right) CD44v6+ and CD44v6– SDCSCs in the presence of 5FU (top) and oxaliplatin (bottom; n = 6 per time point, Mann-Whitney U test). *P < 0.05, **P < 0.01, ***P < 0.001.

Journal: JCI Insight

Article Title: Cancer stem cell–associated miRNAs serve as prognostic biomarkers in colorectal cancer

doi: 10.1172/jci.insight.125294

Figure Lengend Snippet: (A) FACS analysis of CD44v6-positive and -negative sphere-derived cancer stem cells (SDCSCs) derived from a HCT116 cell line. (B) Spheroid-forming capacity of HCT116- and HT29-derived CD44v6+ and CD44v6– SDCSCs (n = 16, Mann-Whitney U test). (C) Spheroid-forming ability of HCT116 (left) and HT29 (right) CD44v6+ and CD44v6– SDCSCs in the presence of 5FU (top) and oxaliplatin (bottom; n = 6 per time point, Mann-Whitney U test). *P < 0.05, **P < 0.01, ***P < 0.001.

Techniques: Derivative Assay, MANN-WHITNEY

(A) Heatmap of differentially expressed miRNAs between CD44v6+ and CD44v6– sphere-derived cancer stem cells (SDCSCs). (B) List of upregulated and downregulated miRNAs ranked by P values. (C) List of pathways identified by KEGG analysis based on predicted downstream target genes for differentially expressed miRNAs between CD44v6+ and CD44v6– SDCSCs. Down, downregulated; Up, upregulated.

Journal: JCI Insight

Article Title: Cancer stem cell–associated miRNAs serve as prognostic biomarkers in colorectal cancer

doi: 10.1172/jci.insight.125294

Figure Lengend Snippet: (A) Heatmap of differentially expressed miRNAs between CD44v6+ and CD44v6– sphere-derived cancer stem cells (SDCSCs). (B) List of upregulated and downregulated miRNAs ranked by P values. (C) List of pathways identified by KEGG analysis based on predicted downstream target genes for differentially expressed miRNAs between CD44v6+ and CD44v6– SDCSCs. Down, downregulated; Up, upregulated.

Techniques: Derivative Assay

(A) Expression of miR-1246 in CD44v6+ and CD44v6– sphere-derived cancer stem cells (SDCSCs) derived from HCT116 and HT29 cell lines (n = 3, Mann-Whitney U test). (B) Expression of miR-1246 in 5FUR and parental HCT116 cell line (n = 3, Mann-Whitney U test). (C) Correlation analysis between CD44v6 and miR-1246 expression in CRC tissue from clinical cohort 1. (D) miR-1246 expression in cancer tissues versus adjacent normal mucosa (left), and in normal tissue and CRC at different stages (right; 1-way ANOVA). (E) Kaplan-Meier analysis of overall survival and disease-free survival in cohort 2 (log-rank test). *P < 0.05, **P < 0.01, ***P < 0.001.

Journal: JCI Insight

Article Title: Cancer stem cell–associated miRNAs serve as prognostic biomarkers in colorectal cancer

doi: 10.1172/jci.insight.125294

Figure Lengend Snippet: (A) Expression of miR-1246 in CD44v6+ and CD44v6– sphere-derived cancer stem cells (SDCSCs) derived from HCT116 and HT29 cell lines (n = 3, Mann-Whitney U test). (B) Expression of miR-1246 in 5FUR and parental HCT116 cell line (n = 3, Mann-Whitney U test). (C) Correlation analysis between CD44v6 and miR-1246 expression in CRC tissue from clinical cohort 1. (D) miR-1246 expression in cancer tissues versus adjacent normal mucosa (left), and in normal tissue and CRC at different stages (right; 1-way ANOVA). (E) Kaplan-Meier analysis of overall survival and disease-free survival in cohort 2 (log-rank test). *P < 0.05, **P < 0.01, ***P < 0.001.

Techniques: Expressing, Derivative Assay, MANN-WHITNEY

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