anti cd24  (Cell Signaling Technology Inc)


Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Cell Signaling Technology Inc anti cd24
    3-Cl-AHPC-mediated inhibition and degradation of pancreatic cancer stem cells spheres of CD133 + , CD44 + <t>/CD24</t> + /EpCAM + , and stem-like spheres of CD44 + <t>/CD24</t> + PANC-1 cells. ((a), (b) and (c), (d)) 3-Cl-AHPC and AHP3 exposure resulted in inhibition of CD44 + /CD24 + /EpCAM + and CD133 + cells growth and sphere formation and degradation of the derived spheres. ((e), (f)) AHP3 and 3-Cl-AHPC inhibited sphere formation and inhibition of growth and degradation of the CD44 + /CD24 + -derived spheres. For sphere formation, the CD44 + /CD24 + /EpCAM + , CD133 + , and CD44 + /CD24 + cells were sorted by flow cytometry and approximately 200–300 cells were seeded with B27 containing DMEM/F12 medium in 96-well low attachment plates and 1 μ M 3-Cl-AHPC or AHP3 added either the day after seeding or 7 days following sphere formation. The sizes of spheres were photographed and measured on a 100 μ m scale and magnification 400X using Olympus fluorescence microscope digital camera software and DP2-BSW software. The error bars represent the mean of 15 sphere determinations ± the standard deviation. ** was significantly different in comparison to control spheres. Data were analyzed by ANOVA, Tukey HSD test for multiple comparisons. ** P < 0.01 versus control.
    Anti Cd24, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti cd24/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti cd24 - by Bioz Stars, 2023-02
    94/100 stars

    Images

    1) Product Images from "Adamantyl Retinoid-Related Molecules Induce Apoptosis in Pancreatic Cancer Cells by Inhibiting IGF-1R and Wnt/ β -Catenin Pathways"

    Article Title: Adamantyl Retinoid-Related Molecules Induce Apoptosis in Pancreatic Cancer Cells by Inhibiting IGF-1R and Wnt/ β -Catenin Pathways

    Journal: Journal of Oncology

    doi: 10.1155/2012/796729

    3-Cl-AHPC-mediated inhibition and degradation of pancreatic cancer stem cells spheres of CD133 + , CD44 + /CD24 + /EpCAM + , and stem-like spheres of CD44 + /CD24 + PANC-1 cells. ((a), (b) and (c), (d)) 3-Cl-AHPC and AHP3 exposure resulted in inhibition of CD44 + /CD24 + /EpCAM + and CD133 + cells growth and sphere formation and degradation of the derived spheres. ((e), (f)) AHP3 and 3-Cl-AHPC inhibited sphere formation and inhibition of growth and degradation of the CD44 + /CD24 + -derived spheres. For sphere formation, the CD44 + /CD24 + /EpCAM + , CD133 + , and CD44 + /CD24 + cells were sorted by flow cytometry and approximately 200–300 cells were seeded with B27 containing DMEM/F12 medium in 96-well low attachment plates and 1 μ M 3-Cl-AHPC or AHP3 added either the day after seeding or 7 days following sphere formation. The sizes of spheres were photographed and measured on a 100 μ m scale and magnification 400X using Olympus fluorescence microscope digital camera software and DP2-BSW software. The error bars represent the mean of 15 sphere determinations ± the standard deviation. ** was significantly different in comparison to control spheres. Data were analyzed by ANOVA, Tukey HSD test for multiple comparisons. ** P < 0.01 versus control.
    Figure Legend Snippet: 3-Cl-AHPC-mediated inhibition and degradation of pancreatic cancer stem cells spheres of CD133 + , CD44 + /CD24 + /EpCAM + , and stem-like spheres of CD44 + /CD24 + PANC-1 cells. ((a), (b) and (c), (d)) 3-Cl-AHPC and AHP3 exposure resulted in inhibition of CD44 + /CD24 + /EpCAM + and CD133 + cells growth and sphere formation and degradation of the derived spheres. ((e), (f)) AHP3 and 3-Cl-AHPC inhibited sphere formation and inhibition of growth and degradation of the CD44 + /CD24 + -derived spheres. For sphere formation, the CD44 + /CD24 + /EpCAM + , CD133 + , and CD44 + /CD24 + cells were sorted by flow cytometry and approximately 200–300 cells were seeded with B27 containing DMEM/F12 medium in 96-well low attachment plates and 1 μ M 3-Cl-AHPC or AHP3 added either the day after seeding or 7 days following sphere formation. The sizes of spheres were photographed and measured on a 100 μ m scale and magnification 400X using Olympus fluorescence microscope digital camera software and DP2-BSW software. The error bars represent the mean of 15 sphere determinations ± the standard deviation. ** was significantly different in comparison to control spheres. Data were analyzed by ANOVA, Tukey HSD test for multiple comparisons. ** P < 0.01 versus control.

    Techniques Used: Inhibition, Derivative Assay, Flow Cytometry, Fluorescence, Microscopy, Software, Standard Deviation

    3-Cl-AHPC- (1 μ M) and AHP3- (1 μ M) mediated inhibition of CD44 + /CD24 + stem-like cell sphere formation and degradation of spheres derived from MiaPaCa-2 and Capan-2 cell lines. 3-Cl-AHPC and AHP3 were added at the time cells were seeded ((a), (c)) or 7 days after cells sphere formation ((b), (d)). The ARR affect on sphere growth was assessed at days 7 and 14 (a), days 7 and 14 (b), days 14 and 21 (c), and days 14 and 21 (d), respectively. The error bars represent the mean of 15 sphere determinations ± the standard deviation. ** Was significantly different in comparison to control spheres. Data were analyzed by ANOVA, Tukey HSD test for multiple comparisons. ** P < 0.01 versus control.
    Figure Legend Snippet: 3-Cl-AHPC- (1 μ M) and AHP3- (1 μ M) mediated inhibition of CD44 + /CD24 + stem-like cell sphere formation and degradation of spheres derived from MiaPaCa-2 and Capan-2 cell lines. 3-Cl-AHPC and AHP3 were added at the time cells were seeded ((a), (c)) or 7 days after cells sphere formation ((b), (d)). The ARR affect on sphere growth was assessed at days 7 and 14 (a), days 7 and 14 (b), days 14 and 21 (c), and days 14 and 21 (d), respectively. The error bars represent the mean of 15 sphere determinations ± the standard deviation. ** Was significantly different in comparison to control spheres. Data were analyzed by ANOVA, Tukey HSD test for multiple comparisons. ** P < 0.01 versus control.

    Techniques Used: Inhibition, Derivative Assay, Standard Deviation

    Dose-response effect of 3-Cl-AHPC on CD44 + /CD24 + cells sphere formation and apoptosis in PANC-1 sphere cells. (a) Addition of 0.25, 0.5, and 1.0 μ M 3-Cl-AHPC added at time of cell seeding inhibited sphere formation at 7 and 14 days. (b) 0.5 and 1.0 μ M 3-Cl-AHPC inhibited sphere formation when added 7 days following sphere formation. (c) 1.0 μ M 3-Cl-AHPC induced apoptosis in CD44 + /CD24 + sphere cells as indicated by nuclear fragmentation detected by acridine orange/ethidium bromide and (d) DAPI staining. Spheres were visualized and photographed utilizing a fluorescence microscope. (e) Apoptosis of sphere cells as demonstrated by TUNEL assay. CD44 + /CD24 + spheres were treated with 1.0 μ M ARRs for 7 days (7D) after sphere formation. Details of slides preparation, visualization, antibodies utilized, and TUNNEL assay methodologies were as described in .
    Figure Legend Snippet: Dose-response effect of 3-Cl-AHPC on CD44 + /CD24 + cells sphere formation and apoptosis in PANC-1 sphere cells. (a) Addition of 0.25, 0.5, and 1.0 μ M 3-Cl-AHPC added at time of cell seeding inhibited sphere formation at 7 and 14 days. (b) 0.5 and 1.0 μ M 3-Cl-AHPC inhibited sphere formation when added 7 days following sphere formation. (c) 1.0 μ M 3-Cl-AHPC induced apoptosis in CD44 + /CD24 + sphere cells as indicated by nuclear fragmentation detected by acridine orange/ethidium bromide and (d) DAPI staining. Spheres were visualized and photographed utilizing a fluorescence microscope. (e) Apoptosis of sphere cells as demonstrated by TUNEL assay. CD44 + /CD24 + spheres were treated with 1.0 μ M ARRs for 7 days (7D) after sphere formation. Details of slides preparation, visualization, antibodies utilized, and TUNNEL assay methodologies were as described in .

    Techniques Used: Staining, Fluorescence, Microscopy, TUNEL Assay

    3-Cl-AHPC and AHP induced apoptosis in PANC-1 CD44 + /CD24 + cells and 3-Cl-AHPC decreased expression of IGF-1R, cyclin D1, and β -catenin in pancreatic cancer cells. (a) Percentage of total apoptotic cells. (b) Percentage of CD44 + /CD24 + cells in the early (Annexin V-FITC positive and PI negative) or late (Annexin V-FITC positive and PI positive) apoptotic cell populations. (c) Percentage of total CD44 + /CD24 + apoptotic cells (Annexin V-FITC positive and PI positive). Cells were treated with 1.0 μ M 3-Cl-AHPC and AHP3 for 96 h. Antibody-conjugated markers CD44-APC-Cy7, CD24-APC, Annexin V-FITC, and PI were used to detect apoptotic and CD44 + /CD24 + cells from the same samples. The error bars represent the mean of three separate determinations ± the standard deviation. ((d), (e)) IGF-1R, cyclin D1, and β -catenin expression decreased following 3-Cl-AHPC exposure in pancreatic cancer cells.
    Figure Legend Snippet: 3-Cl-AHPC and AHP induced apoptosis in PANC-1 CD44 + /CD24 + cells and 3-Cl-AHPC decreased expression of IGF-1R, cyclin D1, and β -catenin in pancreatic cancer cells. (a) Percentage of total apoptotic cells. (b) Percentage of CD44 + /CD24 + cells in the early (Annexin V-FITC positive and PI negative) or late (Annexin V-FITC positive and PI positive) apoptotic cell populations. (c) Percentage of total CD44 + /CD24 + apoptotic cells (Annexin V-FITC positive and PI positive). Cells were treated with 1.0 μ M 3-Cl-AHPC and AHP3 for 96 h. Antibody-conjugated markers CD44-APC-Cy7, CD24-APC, Annexin V-FITC, and PI were used to detect apoptotic and CD44 + /CD24 + cells from the same samples. The error bars represent the mean of three separate determinations ± the standard deviation. ((d), (e)) IGF-1R, cyclin D1, and β -catenin expression decreased following 3-Cl-AHPC exposure in pancreatic cancer cells.

    Techniques Used: Expressing, Standard Deviation

    3-Cl-AHPC decreased expression of IGF-1R, cyclin D1, β -catenin, and cleaved Notch-1 and increased levels of cleaved-caspase-3 in CD44 + /CD24 + spheres. (a) IGF-1R, cyclin D1, and β -catenin expression decreased cleaved-caspase-3 increased with no change in Notch-1 protein levels in spheres following exposure to 3-Cl-AHPC. Pancreatic cancer cells and PANC-1 spheres were exposed to 1.0 μ M 3-Cl-AHPC for 7 days. Western blots were prepared as described in Materials and Methods. (b) mRNA expression of GLI1, GLI2, and Ptch1 in PANC-1 cells. Cells were grown in the presence of 1 μ M 3-Cl-AHPC or vehicle alone (control). (c) Knockdown of IGF-1R expression by sh-IGF-1R inhibited sphere formation and enhanced ARR inhibition of sphere formation. The error bars represent the mean of three separate determinations +/− the standard deviation. •• was significantly different between spheres comprised of sh-vector cells treated with vehicle and 3-Cl-AHPC or AHP3. ♦♦ was significantly different between spheres comprised of sh-vector and IGF-1R-KD1 or IGF-1R-KD 2 at 7 and 14 days, respectively. ** was significantly different in comparison between IGF-1R-KDl/IGF-1R-KD2 spheres (vehicle treated) and IGF-1R-KD1/IGF-1R-KD2 spheres treated with 3-Cl-AHPC or AHP3. Data were analyzed by ANOVA, Tukey HSD test for multiple comparisons, ♦♦, ••, and ** P < 0.01. (d) Knockdown (KD) of IGF-1R enhanced AHP3- and 3-Cl-AHPC-mediated apoptosis in the PANC-1 cells and IGF-IR protein expression in IGF-1R knockdown cells. Apoptosis was assessed using acridine orange/ethidium bromide staining as described in .
    Figure Legend Snippet: 3-Cl-AHPC decreased expression of IGF-1R, cyclin D1, β -catenin, and cleaved Notch-1 and increased levels of cleaved-caspase-3 in CD44 + /CD24 + spheres. (a) IGF-1R, cyclin D1, and β -catenin expression decreased cleaved-caspase-3 increased with no change in Notch-1 protein levels in spheres following exposure to 3-Cl-AHPC. Pancreatic cancer cells and PANC-1 spheres were exposed to 1.0 μ M 3-Cl-AHPC for 7 days. Western blots were prepared as described in Materials and Methods. (b) mRNA expression of GLI1, GLI2, and Ptch1 in PANC-1 cells. Cells were grown in the presence of 1 μ M 3-Cl-AHPC or vehicle alone (control). (c) Knockdown of IGF-1R expression by sh-IGF-1R inhibited sphere formation and enhanced ARR inhibition of sphere formation. The error bars represent the mean of three separate determinations +/− the standard deviation. •• was significantly different between spheres comprised of sh-vector cells treated with vehicle and 3-Cl-AHPC or AHP3. ♦♦ was significantly different between spheres comprised of sh-vector and IGF-1R-KD1 or IGF-1R-KD 2 at 7 and 14 days, respectively. ** was significantly different in comparison between IGF-1R-KDl/IGF-1R-KD2 spheres (vehicle treated) and IGF-1R-KD1/IGF-1R-KD2 spheres treated with 3-Cl-AHPC or AHP3. Data were analyzed by ANOVA, Tukey HSD test for multiple comparisons, ♦♦, ••, and ** P < 0.01. (d) Knockdown (KD) of IGF-1R enhanced AHP3- and 3-Cl-AHPC-mediated apoptosis in the PANC-1 cells and IGF-IR protein expression in IGF-1R knockdown cells. Apoptosis was assessed using acridine orange/ethidium bromide staining as described in .

    Techniques Used: Expressing, Western Blot, Inhibition, Standard Deviation, Plasmid Preparation, Staining

    3-Cl-AHPC mediated inhibition of the activation of TCF/LEF in Wnt/ β -catenin pathway and decreased of β -catenin nuclear localization. (a) 3-Cl-AHPC decreased nuclear β -catenin as indicated by Western blot using nuclear extracts and densitometric quantification. (b) Nuclear β -catenin in control- (i) and 3-Cl-AHPC- (ii) treated PANC-1 cells using confocal fluorescent microscope (magnification 40X). Cells were grown in eight chambered slides and then treated with 3-Cl-AHPC for 24 h. Slide was prepared as described in . DAPI was used for nuclear staining for 1 min and mounted the slide with prolong gold antifade kit. (c) 3-Cl-AHPC inhibited TCF/LEF activity in Wnt/ β -catenin signaling in stably transfected Cignal TCF/LEF-Luc reporter PANC-1 cell lines and 50 mM LiCl was used as a positive control. For CD44/CD24 cells, TCF/LEF stably transfected cells were sorted by flow cytometry and followed the procedure same as wild type (Wt) stable cell line. Luciferase promoter activity values are expressed as fold using a total protein concentration for internal normalization. The error bars represent the mean of three separate determinations ± the standard deviation (SD). (d) 3-Cl-AHPC decreased Wnt/ β -catenin signaling responsive c-Myc protein. ((e) and (f)) Knock down of β -catenin inhibited cell proliferations and enhanced more apoptosis in sh- β -catenin knockdown (KD) PANC-1 cell lines. Proliferation inhibition was evaluated after 72 h of seeding the cells by MTT assay and expressed as absorbance measured at 570 nm. The error bars represent the mean of three separate determinations ± the standard deviation (SD). ** (<0.01) was significantly different in comparison between sh-vector and Catenin-KDl/Catenin-KD2 and also between sh-vector and catenin-KD1/Catenin-KD2 treated with 3-Cl-AHPC, respectively.
    Figure Legend Snippet: 3-Cl-AHPC mediated inhibition of the activation of TCF/LEF in Wnt/ β -catenin pathway and decreased of β -catenin nuclear localization. (a) 3-Cl-AHPC decreased nuclear β -catenin as indicated by Western blot using nuclear extracts and densitometric quantification. (b) Nuclear β -catenin in control- (i) and 3-Cl-AHPC- (ii) treated PANC-1 cells using confocal fluorescent microscope (magnification 40X). Cells were grown in eight chambered slides and then treated with 3-Cl-AHPC for 24 h. Slide was prepared as described in . DAPI was used for nuclear staining for 1 min and mounted the slide with prolong gold antifade kit. (c) 3-Cl-AHPC inhibited TCF/LEF activity in Wnt/ β -catenin signaling in stably transfected Cignal TCF/LEF-Luc reporter PANC-1 cell lines and 50 mM LiCl was used as a positive control. For CD44/CD24 cells, TCF/LEF stably transfected cells were sorted by flow cytometry and followed the procedure same as wild type (Wt) stable cell line. Luciferase promoter activity values are expressed as fold using a total protein concentration for internal normalization. The error bars represent the mean of three separate determinations ± the standard deviation (SD). (d) 3-Cl-AHPC decreased Wnt/ β -catenin signaling responsive c-Myc protein. ((e) and (f)) Knock down of β -catenin inhibited cell proliferations and enhanced more apoptosis in sh- β -catenin knockdown (KD) PANC-1 cell lines. Proliferation inhibition was evaluated after 72 h of seeding the cells by MTT assay and expressed as absorbance measured at 570 nm. The error bars represent the mean of three separate determinations ± the standard deviation (SD). ** (<0.01) was significantly different in comparison between sh-vector and Catenin-KDl/Catenin-KD2 and also between sh-vector and catenin-KD1/Catenin-KD2 treated with 3-Cl-AHPC, respectively.

    Techniques Used: Inhibition, Activation Assay, Western Blot, Microscopy, Staining, Activity Assay, Stable Transfection, Transfection, Positive Control, Flow Cytometry, Luciferase, Protein Concentration, Standard Deviation, MTT Assay, Plasmid Preparation

    anti cd24  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Cell Signaling Technology Inc anti cd24
    3-Cl-AHPC-mediated inhibition and degradation of pancreatic cancer stem cells spheres of CD133 + , CD44 + <t>/CD24</t> + /EpCAM + , and stem-like spheres of CD44 + <t>/CD24</t> + PANC-1 cells. ((a), (b) and (c), (d)) 3-Cl-AHPC and AHP3 exposure resulted in inhibition of CD44 + /CD24 + /EpCAM + and CD133 + cells growth and sphere formation and degradation of the derived spheres. ((e), (f)) AHP3 and 3-Cl-AHPC inhibited sphere formation and inhibition of growth and degradation of the CD44 + /CD24 + -derived spheres. For sphere formation, the CD44 + /CD24 + /EpCAM + , CD133 + , and CD44 + /CD24 + cells were sorted by flow cytometry and approximately 200–300 cells were seeded with B27 containing DMEM/F12 medium in 96-well low attachment plates and 1 μ M 3-Cl-AHPC or AHP3 added either the day after seeding or 7 days following sphere formation. The sizes of spheres were photographed and measured on a 100 μ m scale and magnification 400X using Olympus fluorescence microscope digital camera software and DP2-BSW software. The error bars represent the mean of 15 sphere determinations ± the standard deviation. ** was significantly different in comparison to control spheres. Data were analyzed by ANOVA, Tukey HSD test for multiple comparisons. ** P < 0.01 versus control.
    Anti Cd24, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti cd24/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti cd24 - by Bioz Stars, 2023-02
    94/100 stars

    Images

    1) Product Images from "Adamantyl Retinoid-Related Molecules Induce Apoptosis in Pancreatic Cancer Cells by Inhibiting IGF-1R and Wnt/ β -Catenin Pathways"

    Article Title: Adamantyl Retinoid-Related Molecules Induce Apoptosis in Pancreatic Cancer Cells by Inhibiting IGF-1R and Wnt/ β -Catenin Pathways

    Journal: Journal of Oncology

    doi: 10.1155/2012/796729

    3-Cl-AHPC-mediated inhibition and degradation of pancreatic cancer stem cells spheres of CD133 + , CD44 + /CD24 + /EpCAM + , and stem-like spheres of CD44 + /CD24 + PANC-1 cells. ((a), (b) and (c), (d)) 3-Cl-AHPC and AHP3 exposure resulted in inhibition of CD44 + /CD24 + /EpCAM + and CD133 + cells growth and sphere formation and degradation of the derived spheres. ((e), (f)) AHP3 and 3-Cl-AHPC inhibited sphere formation and inhibition of growth and degradation of the CD44 + /CD24 + -derived spheres. For sphere formation, the CD44 + /CD24 + /EpCAM + , CD133 + , and CD44 + /CD24 + cells were sorted by flow cytometry and approximately 200–300 cells were seeded with B27 containing DMEM/F12 medium in 96-well low attachment plates and 1 μ M 3-Cl-AHPC or AHP3 added either the day after seeding or 7 days following sphere formation. The sizes of spheres were photographed and measured on a 100 μ m scale and magnification 400X using Olympus fluorescence microscope digital camera software and DP2-BSW software. The error bars represent the mean of 15 sphere determinations ± the standard deviation. ** was significantly different in comparison to control spheres. Data were analyzed by ANOVA, Tukey HSD test for multiple comparisons. ** P < 0.01 versus control.
    Figure Legend Snippet: 3-Cl-AHPC-mediated inhibition and degradation of pancreatic cancer stem cells spheres of CD133 + , CD44 + /CD24 + /EpCAM + , and stem-like spheres of CD44 + /CD24 + PANC-1 cells. ((a), (b) and (c), (d)) 3-Cl-AHPC and AHP3 exposure resulted in inhibition of CD44 + /CD24 + /EpCAM + and CD133 + cells growth and sphere formation and degradation of the derived spheres. ((e), (f)) AHP3 and 3-Cl-AHPC inhibited sphere formation and inhibition of growth and degradation of the CD44 + /CD24 + -derived spheres. For sphere formation, the CD44 + /CD24 + /EpCAM + , CD133 + , and CD44 + /CD24 + cells were sorted by flow cytometry and approximately 200–300 cells were seeded with B27 containing DMEM/F12 medium in 96-well low attachment plates and 1 μ M 3-Cl-AHPC or AHP3 added either the day after seeding or 7 days following sphere formation. The sizes of spheres were photographed and measured on a 100 μ m scale and magnification 400X using Olympus fluorescence microscope digital camera software and DP2-BSW software. The error bars represent the mean of 15 sphere determinations ± the standard deviation. ** was significantly different in comparison to control spheres. Data were analyzed by ANOVA, Tukey HSD test for multiple comparisons. ** P < 0.01 versus control.

    Techniques Used: Inhibition, Derivative Assay, Flow Cytometry, Fluorescence, Microscopy, Software, Standard Deviation

    3-Cl-AHPC- (1 μ M) and AHP3- (1 μ M) mediated inhibition of CD44 + /CD24 + stem-like cell sphere formation and degradation of spheres derived from MiaPaCa-2 and Capan-2 cell lines. 3-Cl-AHPC and AHP3 were added at the time cells were seeded ((a), (c)) or 7 days after cells sphere formation ((b), (d)). The ARR affect on sphere growth was assessed at days 7 and 14 (a), days 7 and 14 (b), days 14 and 21 (c), and days 14 and 21 (d), respectively. The error bars represent the mean of 15 sphere determinations ± the standard deviation. ** Was significantly different in comparison to control spheres. Data were analyzed by ANOVA, Tukey HSD test for multiple comparisons. ** P < 0.01 versus control.
    Figure Legend Snippet: 3-Cl-AHPC- (1 μ M) and AHP3- (1 μ M) mediated inhibition of CD44 + /CD24 + stem-like cell sphere formation and degradation of spheres derived from MiaPaCa-2 and Capan-2 cell lines. 3-Cl-AHPC and AHP3 were added at the time cells were seeded ((a), (c)) or 7 days after cells sphere formation ((b), (d)). The ARR affect on sphere growth was assessed at days 7 and 14 (a), days 7 and 14 (b), days 14 and 21 (c), and days 14 and 21 (d), respectively. The error bars represent the mean of 15 sphere determinations ± the standard deviation. ** Was significantly different in comparison to control spheres. Data were analyzed by ANOVA, Tukey HSD test for multiple comparisons. ** P < 0.01 versus control.

    Techniques Used: Inhibition, Derivative Assay, Standard Deviation

    Dose-response effect of 3-Cl-AHPC on CD44 + /CD24 + cells sphere formation and apoptosis in PANC-1 sphere cells. (a) Addition of 0.25, 0.5, and 1.0 μ M 3-Cl-AHPC added at time of cell seeding inhibited sphere formation at 7 and 14 days. (b) 0.5 and 1.0 μ M 3-Cl-AHPC inhibited sphere formation when added 7 days following sphere formation. (c) 1.0 μ M 3-Cl-AHPC induced apoptosis in CD44 + /CD24 + sphere cells as indicated by nuclear fragmentation detected by acridine orange/ethidium bromide and (d) DAPI staining. Spheres were visualized and photographed utilizing a fluorescence microscope. (e) Apoptosis of sphere cells as demonstrated by TUNEL assay. CD44 + /CD24 + spheres were treated with 1.0 μ M ARRs for 7 days (7D) after sphere formation. Details of slides preparation, visualization, antibodies utilized, and TUNNEL assay methodologies were as described in .
    Figure Legend Snippet: Dose-response effect of 3-Cl-AHPC on CD44 + /CD24 + cells sphere formation and apoptosis in PANC-1 sphere cells. (a) Addition of 0.25, 0.5, and 1.0 μ M 3-Cl-AHPC added at time of cell seeding inhibited sphere formation at 7 and 14 days. (b) 0.5 and 1.0 μ M 3-Cl-AHPC inhibited sphere formation when added 7 days following sphere formation. (c) 1.0 μ M 3-Cl-AHPC induced apoptosis in CD44 + /CD24 + sphere cells as indicated by nuclear fragmentation detected by acridine orange/ethidium bromide and (d) DAPI staining. Spheres were visualized and photographed utilizing a fluorescence microscope. (e) Apoptosis of sphere cells as demonstrated by TUNEL assay. CD44 + /CD24 + spheres were treated with 1.0 μ M ARRs for 7 days (7D) after sphere formation. Details of slides preparation, visualization, antibodies utilized, and TUNNEL assay methodologies were as described in .

    Techniques Used: Staining, Fluorescence, Microscopy, TUNEL Assay

    3-Cl-AHPC and AHP induced apoptosis in PANC-1 CD44 + /CD24 + cells and 3-Cl-AHPC decreased expression of IGF-1R, cyclin D1, and β -catenin in pancreatic cancer cells. (a) Percentage of total apoptotic cells. (b) Percentage of CD44 + /CD24 + cells in the early (Annexin V-FITC positive and PI negative) or late (Annexin V-FITC positive and PI positive) apoptotic cell populations. (c) Percentage of total CD44 + /CD24 + apoptotic cells (Annexin V-FITC positive and PI positive). Cells were treated with 1.0 μ M 3-Cl-AHPC and AHP3 for 96 h. Antibody-conjugated markers CD44-APC-Cy7, CD24-APC, Annexin V-FITC, and PI were used to detect apoptotic and CD44 + /CD24 + cells from the same samples. The error bars represent the mean of three separate determinations ± the standard deviation. ((d), (e)) IGF-1R, cyclin D1, and β -catenin expression decreased following 3-Cl-AHPC exposure in pancreatic cancer cells.
    Figure Legend Snippet: 3-Cl-AHPC and AHP induced apoptosis in PANC-1 CD44 + /CD24 + cells and 3-Cl-AHPC decreased expression of IGF-1R, cyclin D1, and β -catenin in pancreatic cancer cells. (a) Percentage of total apoptotic cells. (b) Percentage of CD44 + /CD24 + cells in the early (Annexin V-FITC positive and PI negative) or late (Annexin V-FITC positive and PI positive) apoptotic cell populations. (c) Percentage of total CD44 + /CD24 + apoptotic cells (Annexin V-FITC positive and PI positive). Cells were treated with 1.0 μ M 3-Cl-AHPC and AHP3 for 96 h. Antibody-conjugated markers CD44-APC-Cy7, CD24-APC, Annexin V-FITC, and PI were used to detect apoptotic and CD44 + /CD24 + cells from the same samples. The error bars represent the mean of three separate determinations ± the standard deviation. ((d), (e)) IGF-1R, cyclin D1, and β -catenin expression decreased following 3-Cl-AHPC exposure in pancreatic cancer cells.

    Techniques Used: Expressing, Standard Deviation

    3-Cl-AHPC decreased expression of IGF-1R, cyclin D1, β -catenin, and cleaved Notch-1 and increased levels of cleaved-caspase-3 in CD44 + /CD24 + spheres. (a) IGF-1R, cyclin D1, and β -catenin expression decreased cleaved-caspase-3 increased with no change in Notch-1 protein levels in spheres following exposure to 3-Cl-AHPC. Pancreatic cancer cells and PANC-1 spheres were exposed to 1.0 μ M 3-Cl-AHPC for 7 days. Western blots were prepared as described in Materials and Methods. (b) mRNA expression of GLI1, GLI2, and Ptch1 in PANC-1 cells. Cells were grown in the presence of 1 μ M 3-Cl-AHPC or vehicle alone (control). (c) Knockdown of IGF-1R expression by sh-IGF-1R inhibited sphere formation and enhanced ARR inhibition of sphere formation. The error bars represent the mean of three separate determinations +/− the standard deviation. •• was significantly different between spheres comprised of sh-vector cells treated with vehicle and 3-Cl-AHPC or AHP3. ♦♦ was significantly different between spheres comprised of sh-vector and IGF-1R-KD1 or IGF-1R-KD 2 at 7 and 14 days, respectively. ** was significantly different in comparison between IGF-1R-KDl/IGF-1R-KD2 spheres (vehicle treated) and IGF-1R-KD1/IGF-1R-KD2 spheres treated with 3-Cl-AHPC or AHP3. Data were analyzed by ANOVA, Tukey HSD test for multiple comparisons, ♦♦, ••, and ** P < 0.01. (d) Knockdown (KD) of IGF-1R enhanced AHP3- and 3-Cl-AHPC-mediated apoptosis in the PANC-1 cells and IGF-IR protein expression in IGF-1R knockdown cells. Apoptosis was assessed using acridine orange/ethidium bromide staining as described in .
    Figure Legend Snippet: 3-Cl-AHPC decreased expression of IGF-1R, cyclin D1, β -catenin, and cleaved Notch-1 and increased levels of cleaved-caspase-3 in CD44 + /CD24 + spheres. (a) IGF-1R, cyclin D1, and β -catenin expression decreased cleaved-caspase-3 increased with no change in Notch-1 protein levels in spheres following exposure to 3-Cl-AHPC. Pancreatic cancer cells and PANC-1 spheres were exposed to 1.0 μ M 3-Cl-AHPC for 7 days. Western blots were prepared as described in Materials and Methods. (b) mRNA expression of GLI1, GLI2, and Ptch1 in PANC-1 cells. Cells were grown in the presence of 1 μ M 3-Cl-AHPC or vehicle alone (control). (c) Knockdown of IGF-1R expression by sh-IGF-1R inhibited sphere formation and enhanced ARR inhibition of sphere formation. The error bars represent the mean of three separate determinations +/− the standard deviation. •• was significantly different between spheres comprised of sh-vector cells treated with vehicle and 3-Cl-AHPC or AHP3. ♦♦ was significantly different between spheres comprised of sh-vector and IGF-1R-KD1 or IGF-1R-KD 2 at 7 and 14 days, respectively. ** was significantly different in comparison between IGF-1R-KDl/IGF-1R-KD2 spheres (vehicle treated) and IGF-1R-KD1/IGF-1R-KD2 spheres treated with 3-Cl-AHPC or AHP3. Data were analyzed by ANOVA, Tukey HSD test for multiple comparisons, ♦♦, ••, and ** P < 0.01. (d) Knockdown (KD) of IGF-1R enhanced AHP3- and 3-Cl-AHPC-mediated apoptosis in the PANC-1 cells and IGF-IR protein expression in IGF-1R knockdown cells. Apoptosis was assessed using acridine orange/ethidium bromide staining as described in .

    Techniques Used: Expressing, Western Blot, Inhibition, Standard Deviation, Plasmid Preparation, Staining

    3-Cl-AHPC mediated inhibition of the activation of TCF/LEF in Wnt/ β -catenin pathway and decreased of β -catenin nuclear localization. (a) 3-Cl-AHPC decreased nuclear β -catenin as indicated by Western blot using nuclear extracts and densitometric quantification. (b) Nuclear β -catenin in control- (i) and 3-Cl-AHPC- (ii) treated PANC-1 cells using confocal fluorescent microscope (magnification 40X). Cells were grown in eight chambered slides and then treated with 3-Cl-AHPC for 24 h. Slide was prepared as described in . DAPI was used for nuclear staining for 1 min and mounted the slide with prolong gold antifade kit. (c) 3-Cl-AHPC inhibited TCF/LEF activity in Wnt/ β -catenin signaling in stably transfected Cignal TCF/LEF-Luc reporter PANC-1 cell lines and 50 mM LiCl was used as a positive control. For CD44/CD24 cells, TCF/LEF stably transfected cells were sorted by flow cytometry and followed the procedure same as wild type (Wt) stable cell line. Luciferase promoter activity values are expressed as fold using a total protein concentration for internal normalization. The error bars represent the mean of three separate determinations ± the standard deviation (SD). (d) 3-Cl-AHPC decreased Wnt/ β -catenin signaling responsive c-Myc protein. ((e) and (f)) Knock down of β -catenin inhibited cell proliferations and enhanced more apoptosis in sh- β -catenin knockdown (KD) PANC-1 cell lines. Proliferation inhibition was evaluated after 72 h of seeding the cells by MTT assay and expressed as absorbance measured at 570 nm. The error bars represent the mean of three separate determinations ± the standard deviation (SD). ** (<0.01) was significantly different in comparison between sh-vector and Catenin-KDl/Catenin-KD2 and also between sh-vector and catenin-KD1/Catenin-KD2 treated with 3-Cl-AHPC, respectively.
    Figure Legend Snippet: 3-Cl-AHPC mediated inhibition of the activation of TCF/LEF in Wnt/ β -catenin pathway and decreased of β -catenin nuclear localization. (a) 3-Cl-AHPC decreased nuclear β -catenin as indicated by Western blot using nuclear extracts and densitometric quantification. (b) Nuclear β -catenin in control- (i) and 3-Cl-AHPC- (ii) treated PANC-1 cells using confocal fluorescent microscope (magnification 40X). Cells were grown in eight chambered slides and then treated with 3-Cl-AHPC for 24 h. Slide was prepared as described in . DAPI was used for nuclear staining for 1 min and mounted the slide with prolong gold antifade kit. (c) 3-Cl-AHPC inhibited TCF/LEF activity in Wnt/ β -catenin signaling in stably transfected Cignal TCF/LEF-Luc reporter PANC-1 cell lines and 50 mM LiCl was used as a positive control. For CD44/CD24 cells, TCF/LEF stably transfected cells were sorted by flow cytometry and followed the procedure same as wild type (Wt) stable cell line. Luciferase promoter activity values are expressed as fold using a total protein concentration for internal normalization. The error bars represent the mean of three separate determinations ± the standard deviation (SD). (d) 3-Cl-AHPC decreased Wnt/ β -catenin signaling responsive c-Myc protein. ((e) and (f)) Knock down of β -catenin inhibited cell proliferations and enhanced more apoptosis in sh- β -catenin knockdown (KD) PANC-1 cell lines. Proliferation inhibition was evaluated after 72 h of seeding the cells by MTT assay and expressed as absorbance measured at 570 nm. The error bars represent the mean of three separate determinations ± the standard deviation (SD). ** (<0.01) was significantly different in comparison between sh-vector and Catenin-KDl/Catenin-KD2 and also between sh-vector and catenin-KD1/Catenin-KD2 treated with 3-Cl-AHPC, respectively.

    Techniques Used: Inhibition, Activation Assay, Western Blot, Microscopy, Staining, Activity Assay, Stable Transfection, Transfection, Positive Control, Flow Cytometry, Luciferase, Protein Concentration, Standard Deviation, MTT Assay, Plasmid Preparation

    mouse monoclonal anti cd24  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Cell Signaling Technology Inc mouse monoclonal anti cd24
    Mouse Monoclonal Anti Cd24, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse monoclonal anti cd24/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse monoclonal anti cd24 - by Bioz Stars, 2023-02
    94/100 stars

    Images

    anti cd24  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Cell Signaling Technology Inc anti cd24
    Expression levels of miR-376c-3p and RAB2A in BCSCs. (A) Flow cytometry analysis of the proportion of <t>CD44+CD24-cells.</t> (B) Protein expression levels of stem cell markers OCT4 and SOX2 in MCF-7 cells and BCSCs. (C) Reverse transcription-quantitative PCR analysis of miR-376c-3p and RAB2A expression levels in MCF-10A cells, MCF-7 cells, non-BCSCs and BCSCs. (D) Protein expression levels of RAB2A in MCF-10A cells, MCF-7 cells, non-BCSCs and BCSCs. ** P<0.01 and *** P<0.001. miR, microRNA; RAB2A, Ras-related protein Rab-2A; BCSC, breast cancer stem cell; CD, cluster of differentiation; OCT4, octamer-binding transcription factor 4; SOX2, SRY-box transcription factor 2.
    Anti Cd24, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti cd24/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti cd24 - by Bioz Stars, 2023-02
    94/100 stars

    Images

    1) Product Images from "miR-376c-3p modulates the properties of breast cancer stem cells by targeting RAB2A"

    Article Title: miR-376c-3p modulates the properties of breast cancer stem cells by targeting RAB2A

    Journal: Experimental and Therapeutic Medicine

    doi: 10.3892/etm.2020.9196

    Expression levels of miR-376c-3p and RAB2A in BCSCs. (A) Flow cytometry analysis of the proportion of CD44+CD24-cells. (B) Protein expression levels of stem cell markers OCT4 and SOX2 in MCF-7 cells and BCSCs. (C) Reverse transcription-quantitative PCR analysis of miR-376c-3p and RAB2A expression levels in MCF-10A cells, MCF-7 cells, non-BCSCs and BCSCs. (D) Protein expression levels of RAB2A in MCF-10A cells, MCF-7 cells, non-BCSCs and BCSCs. ** P<0.01 and *** P<0.001. miR, microRNA; RAB2A, Ras-related protein Rab-2A; BCSC, breast cancer stem cell; CD, cluster of differentiation; OCT4, octamer-binding transcription factor 4; SOX2, SRY-box transcription factor 2.
    Figure Legend Snippet: Expression levels of miR-376c-3p and RAB2A in BCSCs. (A) Flow cytometry analysis of the proportion of CD44+CD24-cells. (B) Protein expression levels of stem cell markers OCT4 and SOX2 in MCF-7 cells and BCSCs. (C) Reverse transcription-quantitative PCR analysis of miR-376c-3p and RAB2A expression levels in MCF-10A cells, MCF-7 cells, non-BCSCs and BCSCs. (D) Protein expression levels of RAB2A in MCF-10A cells, MCF-7 cells, non-BCSCs and BCSCs. ** P<0.01 and *** P<0.001. miR, microRNA; RAB2A, Ras-related protein Rab-2A; BCSC, breast cancer stem cell; CD, cluster of differentiation; OCT4, octamer-binding transcription factor 4; SOX2, SRY-box transcription factor 2.

    Techniques Used: Expressing, Flow Cytometry, Real-time Polymerase Chain Reaction, Binding Assay

    miR-376-3p reduces the stem cell population and downregulates stem cell regulatory proteins by targeting RAB2A. (A) Flow cytometry analysis of the proportion of CD44+CD24-cells. The effect of (B) miR-376c-3p mimic, pcDNA-RAB2A, (C) miR-376c-3p inhibitor and si-RAB2A on RAB2A, OCT4 and SOX2 expression levels. * P<0.05, ** P<0.01 and *** P<0.001. miR, microRNA; RAB2A, Ras-related protein Rab-2A; CD, cluster of differentiation; si, small interfering RNA; OCT4, octamer-binding transcription factor 4; SOX2, SRY-box transcription factor 2; NC, negative control.
    Figure Legend Snippet: miR-376-3p reduces the stem cell population and downregulates stem cell regulatory proteins by targeting RAB2A. (A) Flow cytometry analysis of the proportion of CD44+CD24-cells. The effect of (B) miR-376c-3p mimic, pcDNA-RAB2A, (C) miR-376c-3p inhibitor and si-RAB2A on RAB2A, OCT4 and SOX2 expression levels. * P<0.05, ** P<0.01 and *** P<0.001. miR, microRNA; RAB2A, Ras-related protein Rab-2A; CD, cluster of differentiation; si, small interfering RNA; OCT4, octamer-binding transcription factor 4; SOX2, SRY-box transcription factor 2; NC, negative control.

    Techniques Used: Flow Cytometry, Expressing, Small Interfering RNA, Binding Assay, Negative Control

    cd24  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Cell Signaling Technology Inc cd24
    Cd24, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cd24/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cd24 - by Bioz Stars, 2023-02
    94/100 stars

    Images

    cd24  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Cell Signaling Technology Inc cd24
    a, Flow cytometry plots showing the sorting strategy and the purity of ST0 and ST1 cells after sorting. Shown one representative experiment out of three. b, Flow cytometry plots showing expression of PLZF, T-BET and RORγt in ST1 wild-type cells. Graphs show mean percentage and number ± SEM of the indicated populations. n=7 (PLZF hi , PLZF hi T-BET + , and PLZF lo T-BET + ), or n=3 (PLZF int RORγt + ) independent experiments c, Graphs showing the normalized counts of Bcl6 mRNA in ST0 and ST1 cells based on RNA-seq data. n=3 independent experiments (DESeq, ** P < 0.01). d, Flow cytometry plots showing the expression of BCL-6 and CD69 (left) or BCL-6 and EGR-2 (right) in Tetr + TCRβ + <t>CD24</t> + CD44 − iNKT cells. Shown one representative experiment out of four (CD69) or two (EGR-2).
    Cd24, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cd24/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cd24 - by Bioz Stars, 2023-02
    94/100 stars

    Images

    1) Product Images from "The transcription factor BCL-6 controls early development of innate-like T cells"

    Article Title: The transcription factor BCL-6 controls early development of innate-like T cells

    Journal: Nature immunology

    doi: 10.1038/s41590-020-0737-y

    a, Flow cytometry plots showing the sorting strategy and the purity of ST0 and ST1 cells after sorting. Shown one representative experiment out of three. b, Flow cytometry plots showing expression of PLZF, T-BET and RORγt in ST1 wild-type cells. Graphs show mean percentage and number ± SEM of the indicated populations. n=7 (PLZF hi , PLZF hi T-BET + , and PLZF lo T-BET + ), or n=3 (PLZF int RORγt + ) independent experiments c, Graphs showing the normalized counts of Bcl6 mRNA in ST0 and ST1 cells based on RNA-seq data. n=3 independent experiments (DESeq, ** P < 0.01). d, Flow cytometry plots showing the expression of BCL-6 and CD69 (left) or BCL-6 and EGR-2 (right) in Tetr + TCRβ + CD24 + CD44 − iNKT cells. Shown one representative experiment out of four (CD69) or two (EGR-2).
    Figure Legend Snippet: a, Flow cytometry plots showing the sorting strategy and the purity of ST0 and ST1 cells after sorting. Shown one representative experiment out of three. b, Flow cytometry plots showing expression of PLZF, T-BET and RORγt in ST1 wild-type cells. Graphs show mean percentage and number ± SEM of the indicated populations. n=7 (PLZF hi , PLZF hi T-BET + , and PLZF lo T-BET + ), or n=3 (PLZF int RORγt + ) independent experiments c, Graphs showing the normalized counts of Bcl6 mRNA in ST0 and ST1 cells based on RNA-seq data. n=3 independent experiments (DESeq, ** P < 0.01). d, Flow cytometry plots showing the expression of BCL-6 and CD69 (left) or BCL-6 and EGR-2 (right) in Tetr + TCRβ + CD24 + CD44 − iNKT cells. Shown one representative experiment out of four (CD69) or two (EGR-2).

    Techniques Used: Flow Cytometry, Expressing, RNA Sequencing Assay

    a, Volcano plot showing genes that were differentially expressed between ST0 (sorted as Tetramer + TCRβ + CD24 + CD44 − ) and ST1 (Tetramer + TCRβ + CD24 − CD44 − ) iNKT cells. Lines indicate a 2-fold difference between ST0 and ST1 cells (DESeq, P < 0.05). b, Metascape analysis showing the pathways enriched among the genes that were differentially expressed. Metascape analysis was performed with hypergeometric test coupled with Benjamini-Hochberg P value correction algorithm. c, GSEA analysis of the RNA-seq data showing enrichment of T cell memory-related genes in ST1 cells, compared to ST0 cells. Gene sets were obtained from MSigDB for memory CD8 and memory CD4 (GSE11057) T cells. Normalized enrichment scores (NES) and FDR as implemented by GSEA, based on 1,000 permutations. d,e, Heatmaps showing the expression of select transcription factors ( d, upper) and known ST0-expressed genes ( d, lower), and cytokines, chemokines and their receptors ( e ) in the RNA-seq data. n=3 independent experiments with 4-6 pooled thymi each from 4-5 weeks old wild-type mice. Differential gene expression analysis was performed with the DESeq algorithm. * P < 0.05, ** P < 0.01, *** P < 0.001
    Figure Legend Snippet: a, Volcano plot showing genes that were differentially expressed between ST0 (sorted as Tetramer + TCRβ + CD24 + CD44 − ) and ST1 (Tetramer + TCRβ + CD24 − CD44 − ) iNKT cells. Lines indicate a 2-fold difference between ST0 and ST1 cells (DESeq, P < 0.05). b, Metascape analysis showing the pathways enriched among the genes that were differentially expressed. Metascape analysis was performed with hypergeometric test coupled with Benjamini-Hochberg P value correction algorithm. c, GSEA analysis of the RNA-seq data showing enrichment of T cell memory-related genes in ST1 cells, compared to ST0 cells. Gene sets were obtained from MSigDB for memory CD8 and memory CD4 (GSE11057) T cells. Normalized enrichment scores (NES) and FDR as implemented by GSEA, based on 1,000 permutations. d,e, Heatmaps showing the expression of select transcription factors ( d, upper) and known ST0-expressed genes ( d, lower), and cytokines, chemokines and their receptors ( e ) in the RNA-seq data. n=3 independent experiments with 4-6 pooled thymi each from 4-5 weeks old wild-type mice. Differential gene expression analysis was performed with the DESeq algorithm. * P < 0.05, ** P < 0.01, *** P < 0.001

    Techniques Used: RNA Sequencing Assay, Expressing

    a, Graph showing average Bcl6 mRNA expression in the indicated FACS-sorted populations. n=3 independent experiments with 3-5 pooled thymi each from 4-5 weeks old mice. Data represent mean ± SEM. b, Flow cytometry histograms showing expression of BCL-6 protein in the indicated populations. c, Flow cytometry plots showing expression of BCL-6 and CD24 in CD44 − iNKT cells after magnetic bead-based enrichment. d, BCL-6 expression in pre-DP (CD69 − TCRβ − CD4 + CD8 + ), post-DP (CD69 + TCRβ + CD4 + CD8 + ) and ST0 iNKT cells. e, Graph showing the geometric mean fluorescence intensity (gMFI) of BCL-6 in the indicated thymic populations. Data represent mean ± SEM. n=6 independent experiments. Statistical analysis was performed with two-tailed unpaired t -test ** P < 0.01, *** P < 0.001
    Figure Legend Snippet: a, Graph showing average Bcl6 mRNA expression in the indicated FACS-sorted populations. n=3 independent experiments with 3-5 pooled thymi each from 4-5 weeks old mice. Data represent mean ± SEM. b, Flow cytometry histograms showing expression of BCL-6 protein in the indicated populations. c, Flow cytometry plots showing expression of BCL-6 and CD24 in CD44 − iNKT cells after magnetic bead-based enrichment. d, BCL-6 expression in pre-DP (CD69 − TCRβ − CD4 + CD8 + ), post-DP (CD69 + TCRβ + CD4 + CD8 + ) and ST0 iNKT cells. e, Graph showing the geometric mean fluorescence intensity (gMFI) of BCL-6 in the indicated thymic populations. Data represent mean ± SEM. n=6 independent experiments. Statistical analysis was performed with two-tailed unpaired t -test ** P < 0.01, *** P < 0.001

    Techniques Used: Expressing, Flow Cytometry, Fluorescence, Two Tailed Test

    a, Thymocytes from 3-6 weeks old control and Bcl6 Δ/Δ mice were enriched for iNKT cells with magnetic beads and were subsequently stained with anti-TCRβ, anti-CD24, anti-CD44 and anti-NK1.1 to identify the various iNKT developmental stages (ST0-3). Numbers in the plots indicate the percentage of the respective populations. b, Graphs show the absolute cell number and percentage of ST0 cells in control and Bcl6 Δ/Δ mice. Data represent mean ± SEM. n=4 independent experiments. c, Graphs showing the absolute cell number and percentage of ST1, ST2 and ST3 cells in control and Bcl6 Δ/Δ mice. Data represent mean ± SEM. n=6 independent experiments with multiple mice for each experiment. d,e, Flow cytometry plots showing the percentage of ( d ) ST0 and ( e ) ST1-3 cells among CD45.1 + (Comp) and CD45.2 + ( Bcl6 Δ/Δ ) iNKT cells. Graphs show the mean percentage ± SEM in the indicated populations. n=4 (ST0), or n=5 (ST1-3) independent experiments. Statistical analysis was performed with two-tailed unpaired t -test. * P < 0.05, *** P < 0.001
    Figure Legend Snippet: a, Thymocytes from 3-6 weeks old control and Bcl6 Δ/Δ mice were enriched for iNKT cells with magnetic beads and were subsequently stained with anti-TCRβ, anti-CD24, anti-CD44 and anti-NK1.1 to identify the various iNKT developmental stages (ST0-3). Numbers in the plots indicate the percentage of the respective populations. b, Graphs show the absolute cell number and percentage of ST0 cells in control and Bcl6 Δ/Δ mice. Data represent mean ± SEM. n=4 independent experiments. c, Graphs showing the absolute cell number and percentage of ST1, ST2 and ST3 cells in control and Bcl6 Δ/Δ mice. Data represent mean ± SEM. n=6 independent experiments with multiple mice for each experiment. d,e, Flow cytometry plots showing the percentage of ( d ) ST0 and ( e ) ST1-3 cells among CD45.1 + (Comp) and CD45.2 + ( Bcl6 Δ/Δ ) iNKT cells. Graphs show the mean percentage ± SEM in the indicated populations. n=4 (ST0), or n=5 (ST1-3) independent experiments. Statistical analysis was performed with two-tailed unpaired t -test. * P < 0.05, *** P < 0.001

    Techniques Used: Magnetic Beads, Staining, Flow Cytometry, Two Tailed Test

    cd24  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Cell Signaling Technology Inc cd24
    Cd24, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cd24/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cd24 - by Bioz Stars, 2023-02
    94/100 stars

    Images

    cd24  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Cell Signaling Technology Inc cd24
    Altered expression of <t>CD24</t> does not alter the viability or spontaneous apoptosis of CRC cells. HCT116 and HT29 CRC cells were transfected with the control plasmid (CD24VEC, CD24NC) or the CD24OE plasmid or CD24-specific short hairpin RNA (CD24KD). (A) Western blot analysis of the expression of CD24 to demonstrate overexpression of CD24 in HCT116 cells and CD24-silencing in HT29 cells. Western blotting was performed for analysis of the relative levels of c-caspase-3 and c-PARP in individual groups of CRC cells. (B) Impact of altered expression of CD24 on cell viability was determined using cell counting kit-8 assays. (C) Percentages of apoptotic cells were determined by Hoechst 33258 staining and quantitatively analyzed. Data are representative images (magnification, ×200) or are expressed as the mean ± SD of each group of cells from three separate experiments. CRC, colorectal cancer; VEC, vector control; NC, negative control; OE, overexpression; KD, knockdown; PARP, poly(ADP-ribose) polymerase; c-, cleaved.
    Cd24, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cd24/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cd24 - by Bioz Stars, 2023-02
    94/100 stars

    Images

    1) Product Images from "Combination of targeting CD24 and inhibiting autophagy suppresses the proliferation and enhances the apoptosis of colorectal cancer cells"

    Article Title: Combination of targeting CD24 and inhibiting autophagy suppresses the proliferation and enhances the apoptosis of colorectal cancer cells

    Journal: Molecular Medicine Reports

    doi: 10.3892/mmr.2019.10288

    Altered expression of CD24 does not alter the viability or spontaneous apoptosis of CRC cells. HCT116 and HT29 CRC cells were transfected with the control plasmid (CD24VEC, CD24NC) or the CD24OE plasmid or CD24-specific short hairpin RNA (CD24KD). (A) Western blot analysis of the expression of CD24 to demonstrate overexpression of CD24 in HCT116 cells and CD24-silencing in HT29 cells. Western blotting was performed for analysis of the relative levels of c-caspase-3 and c-PARP in individual groups of CRC cells. (B) Impact of altered expression of CD24 on cell viability was determined using cell counting kit-8 assays. (C) Percentages of apoptotic cells were determined by Hoechst 33258 staining and quantitatively analyzed. Data are representative images (magnification, ×200) or are expressed as the mean ± SD of each group of cells from three separate experiments. CRC, colorectal cancer; VEC, vector control; NC, negative control; OE, overexpression; KD, knockdown; PARP, poly(ADP-ribose) polymerase; c-, cleaved.
    Figure Legend Snippet: Altered expression of CD24 does not alter the viability or spontaneous apoptosis of CRC cells. HCT116 and HT29 CRC cells were transfected with the control plasmid (CD24VEC, CD24NC) or the CD24OE plasmid or CD24-specific short hairpin RNA (CD24KD). (A) Western blot analysis of the expression of CD24 to demonstrate overexpression of CD24 in HCT116 cells and CD24-silencing in HT29 cells. Western blotting was performed for analysis of the relative levels of c-caspase-3 and c-PARP in individual groups of CRC cells. (B) Impact of altered expression of CD24 on cell viability was determined using cell counting kit-8 assays. (C) Percentages of apoptotic cells were determined by Hoechst 33258 staining and quantitatively analyzed. Data are representative images (magnification, ×200) or are expressed as the mean ± SD of each group of cells from three separate experiments. CRC, colorectal cancer; VEC, vector control; NC, negative control; OE, overexpression; KD, knockdown; PARP, poly(ADP-ribose) polymerase; c-, cleaved.

    Techniques Used: Expressing, Transfection, Plasmid Preparation, shRNA, Western Blot, Over Expression, Cell Counting, Staining, Negative Control

    Altered expression of CD24 modulates autophagy in CRC cells. (A) Western blot analysis of the relative levels of Beclin-1, Atg5 and Atg3 in individual groups of cells. (B) Western blot analysis of the expression of LC3A and LC3B in individual groups of cells. (C) Western blot analysis of the dynamic changes in the relative expression levels of p62 in CD24OE:HCT116 and CD24KD:HT29 cells. (D) Immunofluorescent analysis of punctate LC3B expression in CRC cells. (E) Quantitative analysis of punctate cells. Data are representative images (magnification, ×200) or are expressed as the mean ± SD of each group of cells from three separate experiments. **P<0.01 vs. control. CRC, colorectal cancer; VEC, vector control; NC, negative control; OE, overexpression; KD, knockdown; LC3, microtubule-associated protein-1 light chain-3; Atg3, autophagy-related 3; Atg5, autophagy-related 5; 3-MA, 3-methyladenine.
    Figure Legend Snippet: Altered expression of CD24 modulates autophagy in CRC cells. (A) Western blot analysis of the relative levels of Beclin-1, Atg5 and Atg3 in individual groups of cells. (B) Western blot analysis of the expression of LC3A and LC3B in individual groups of cells. (C) Western blot analysis of the dynamic changes in the relative expression levels of p62 in CD24OE:HCT116 and CD24KD:HT29 cells. (D) Immunofluorescent analysis of punctate LC3B expression in CRC cells. (E) Quantitative analysis of punctate cells. Data are representative images (magnification, ×200) or are expressed as the mean ± SD of each group of cells from three separate experiments. **P<0.01 vs. control. CRC, colorectal cancer; VEC, vector control; NC, negative control; OE, overexpression; KD, knockdown; LC3, microtubule-associated protein-1 light chain-3; Atg3, autophagy-related 3; Atg5, autophagy-related 5; 3-MA, 3-methyladenine.

    Techniques Used: Expressing, Western Blot, Plasmid Preparation, Negative Control, Over Expression

    Inhibition of autophagy by Atg5 silencing modulates the altered expression of CD24-regulated cell viability and apoptosis in colorectal cancer cells. (A) HCT116 and HT29 cells were transfected Atg-specific siRNA or control siRNA and the relative expression levels of Atg5 were determined by a western blot assay. (B) CD24VEC:HCT116, CD24OE:HCT116, CD24NC:HT29 and CD24KD:HT29 cells were transfected Atg5-specific siRNA or control siRNA and the relative expression levels of Beclin-1, Atg5, cleaved PARP and cleaved caspase-3 were determined by western blot analysis. (C) Viabilities of individual groups of cells were determined using cell counting kit-8 assays. (D) Apoptotic cells were examined by Hoechst 33258 staining. (E) Quantitative analysis of apoptotic cells. Data are representative images (magnification, ×200) or are expressed as the mean ± SD of each group of cells from three separate experiments. *P<0.05 and **P<0.01 vs. corresponding control cells. VEC, vector control; NC, negative control; OE, overexpression; KD, knockdown; siRNA, small interfering RNA; PARP, poly (ADP-ribose) polymerase; c-, cleaved; Atg, autophagy-related 5.
    Figure Legend Snippet: Inhibition of autophagy by Atg5 silencing modulates the altered expression of CD24-regulated cell viability and apoptosis in colorectal cancer cells. (A) HCT116 and HT29 cells were transfected Atg-specific siRNA or control siRNA and the relative expression levels of Atg5 were determined by a western blot assay. (B) CD24VEC:HCT116, CD24OE:HCT116, CD24NC:HT29 and CD24KD:HT29 cells were transfected Atg5-specific siRNA or control siRNA and the relative expression levels of Beclin-1, Atg5, cleaved PARP and cleaved caspase-3 were determined by western blot analysis. (C) Viabilities of individual groups of cells were determined using cell counting kit-8 assays. (D) Apoptotic cells were examined by Hoechst 33258 staining. (E) Quantitative analysis of apoptotic cells. Data are representative images (magnification, ×200) or are expressed as the mean ± SD of each group of cells from three separate experiments. *P<0.05 and **P<0.01 vs. corresponding control cells. VEC, vector control; NC, negative control; OE, overexpression; KD, knockdown; siRNA, small interfering RNA; PARP, poly (ADP-ribose) polymerase; c-, cleaved; Atg, autophagy-related 5.

    Techniques Used: Inhibition, Expressing, Transfection, Western Blot, Cell Counting, Staining, Plasmid Preparation, Negative Control, Over Expression, Small Interfering RNA

    Autophagy regulated by altered expression of CD24 is partially regulated by activating NF-κBp65 signaling in colorectal cancer cells. (A) Western blot analysis of the relative levels of NF-κBp65 and IκB-α and p-NF-κBp65 in individual groups of cells. (B) Western blot analysis of p-NF-κBp65, Beclin-1, Atg5 and LC3B in individual groups of cells treated with vehicle DMSO or 5 µM BAY for 24 h. (C) Punctate LC3B expression was determined by immunofluorescence and (D) quantitatively analyzed for cells with punctate LC3B. Data are representative images (magnification, ×200) or are expressed as the mean ± SD of each group of cells from three separate experiments. **P<0.01 vs. untreated corresponding cells. VEC, vector control; NC, negative control; OE, overexpression; KD, knockdown; BAY, Bay11-7082; NF-κB, nuclear factor-κB; IκB-α, inhibitor of NF-κBα; p-, phosphorylated; Atg5, autophagy-related 5; LC3B, microtubule-associated protein-1 light chain-3B.
    Figure Legend Snippet: Autophagy regulated by altered expression of CD24 is partially regulated by activating NF-κBp65 signaling in colorectal cancer cells. (A) Western blot analysis of the relative levels of NF-κBp65 and IκB-α and p-NF-κBp65 in individual groups of cells. (B) Western blot analysis of p-NF-κBp65, Beclin-1, Atg5 and LC3B in individual groups of cells treated with vehicle DMSO or 5 µM BAY for 24 h. (C) Punctate LC3B expression was determined by immunofluorescence and (D) quantitatively analyzed for cells with punctate LC3B. Data are representative images (magnification, ×200) or are expressed as the mean ± SD of each group of cells from three separate experiments. **P<0.01 vs. untreated corresponding cells. VEC, vector control; NC, negative control; OE, overexpression; KD, knockdown; BAY, Bay11-7082; NF-κB, nuclear factor-κB; IκB-α, inhibitor of NF-κBα; p-, phosphorylated; Atg5, autophagy-related 5; LC3B, microtubule-associated protein-1 light chain-3B.

    Techniques Used: Expressing, Western Blot, Immunofluorescence, Plasmid Preparation, Negative Control, Over Expression

    Diagram illustration of the potential roles of CD24 in the development of CRC. CD24 is expressed on the membrane of CRC cells via a GPI-anchor. Over-expression of CD24 induces NF-kBp65 activation to inhibit autophagy in CRC cells, and its impact on CRC cell proliferation and apoptosis depends on the expression levels of CD24. White arrows indicate the effects on cell proliferation, apoptosis and autophagy of altered expression of CD24; black arrows represent the effects on cell proliferation, apoptosis and autophagy of combination treatment of targeting CD24 and inhibiting autophagy. NF-κB, nuclear factor-κB; Atg5, autophagy-related 5; siRNA, small interfering RNA; 3-MA, 3-methyladenine.
    Figure Legend Snippet: Diagram illustration of the potential roles of CD24 in the development of CRC. CD24 is expressed on the membrane of CRC cells via a GPI-anchor. Over-expression of CD24 induces NF-kBp65 activation to inhibit autophagy in CRC cells, and its impact on CRC cell proliferation and apoptosis depends on the expression levels of CD24. White arrows indicate the effects on cell proliferation, apoptosis and autophagy of altered expression of CD24; black arrows represent the effects on cell proliferation, apoptosis and autophagy of combination treatment of targeting CD24 and inhibiting autophagy. NF-κB, nuclear factor-κB; Atg5, autophagy-related 5; siRNA, small interfering RNA; 3-MA, 3-methyladenine.

    Techniques Used: Over Expression, Activation Assay, Expressing, Small Interfering RNA

    mouse monoclonal anti cd24  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Cell Signaling Technology Inc mouse monoclonal anti cd24
    Mouse Monoclonal Anti Cd24, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse monoclonal anti cd24/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse monoclonal anti cd24 - by Bioz Stars, 2023-02
    94/100 stars

    Images

    cd24  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Cell Signaling Technology Inc cd24
    α‐Mangostin inhibits cell proliferation and induces apoptosis in pancreatic cancer stem cells (CSCs) and cell lines but has no effect on human pancreatic normal ductal epithelial (HPNE) cells. (A‐D), <t>CD44+CD24+ESA+CSCs</t> were isolated from human primary pancreatic tumours. Pancreatic CSCs, cancer cell lines (AsPC‐1 and PANC‐1) and (HPNE) cells were treated with α‐Mangostin (0‐10 µmol/L) for 48 h, and cell proliferation was measured by trypan blue assay. (E‐G), Pancreatic CSCs and cancer cell lines (AsPC‐1 and PANC‐1) were treated with α‐Mangostin (0‐10 µmol/L) for 48 h, and apoptosis was measured by TUNEL assay. Data represent mean (n = 4) ± SD. *, #, % and @ = significantly different from control, and each other, P < 0.05
    Cd24, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cd24/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cd24 - by Bioz Stars, 2023-02
    94/100 stars

    Images

    1) Product Images from "Inhibition of pancreatic cancer stem cell characteristics by α‐Mangostin: Molecular mechanisms involving Sonic hedgehog and Nanog"

    Article Title: Inhibition of pancreatic cancer stem cell characteristics by α‐Mangostin: Molecular mechanisms involving Sonic hedgehog and Nanog

    Journal: Journal of Cellular and Molecular Medicine

    doi: 10.1111/jcmm.14178

    α‐Mangostin inhibits cell proliferation and induces apoptosis in pancreatic cancer stem cells (CSCs) and cell lines but has no effect on human pancreatic normal ductal epithelial (HPNE) cells. (A‐D), CD44+CD24+ESA+CSCs were isolated from human primary pancreatic tumours. Pancreatic CSCs, cancer cell lines (AsPC‐1 and PANC‐1) and (HPNE) cells were treated with α‐Mangostin (0‐10 µmol/L) for 48 h, and cell proliferation was measured by trypan blue assay. (E‐G), Pancreatic CSCs and cancer cell lines (AsPC‐1 and PANC‐1) were treated with α‐Mangostin (0‐10 µmol/L) for 48 h, and apoptosis was measured by TUNEL assay. Data represent mean (n = 4) ± SD. *, #, % and @ = significantly different from control, and each other, P < 0.05
    Figure Legend Snippet: α‐Mangostin inhibits cell proliferation and induces apoptosis in pancreatic cancer stem cells (CSCs) and cell lines but has no effect on human pancreatic normal ductal epithelial (HPNE) cells. (A‐D), CD44+CD24+ESA+CSCs were isolated from human primary pancreatic tumours. Pancreatic CSCs, cancer cell lines (AsPC‐1 and PANC‐1) and (HPNE) cells were treated with α‐Mangostin (0‐10 µmol/L) for 48 h, and cell proliferation was measured by trypan blue assay. (E‐G), Pancreatic CSCs and cancer cell lines (AsPC‐1 and PANC‐1) were treated with α‐Mangostin (0‐10 µmol/L) for 48 h, and apoptosis was measured by TUNEL assay. Data represent mean (n = 4) ± SD. *, #, % and @ = significantly different from control, and each other, P < 0.05

    Techniques Used: Isolation, TUNEL Assay

    α‐Mangostin inhibits cell viability in spheroids, and colony formation and the expression of stem cell markers and pluripotency maintain factors by CSCs from pancreatic tumours. (A and B), CSCs were isolated from pancreatic tumours of humans and KrasG12D mice and treated with α‐Mangostin (0‐10 µmol/L) for 7 d to obtain primary spheroids. At the end of the incubation period for a week, the spheroids were collected, reseeded and treated with α‐Mangostin for another week to obtain secondary spheroids. Further, secondary spheroids were collected, reseeded and treated with α‐Mangostin for another week to obtain tertiary spheroids. Cell viability in the spheroids was measured by trypan blue assay at the end of 7, 14 and 21 d. Data represent mean ± SD. *, &, #, %, ˄ and ** = significantly different from control, P < 0.05. C, In soft agar, human pancreatic CSCs were seeded and treated with α‐Mangostin (0‐10 μmol/L) for 21 d. At the end of the incubation period, the number of colonies was counted. *, #, % and @ = significantly different from control (n = 4), P < 0.05. D, Human pancreatic CSCs were treated with α‐Mangostin (0‐10 μmol/L) for 48 h. The expression of CD24, CD44 and CD133 was measured by the Western blot analysis. E, human pancreatic CSCs were treated with α‐Mangostin (0‐10 μmol/L) for 48 h, and the expression of Nanog, Oct4, Sox2, KLF4 and c‐Myc was measured by the Western blot analysis. β‐actin was used as a loading control
    Figure Legend Snippet: α‐Mangostin inhibits cell viability in spheroids, and colony formation and the expression of stem cell markers and pluripotency maintain factors by CSCs from pancreatic tumours. (A and B), CSCs were isolated from pancreatic tumours of humans and KrasG12D mice and treated with α‐Mangostin (0‐10 µmol/L) for 7 d to obtain primary spheroids. At the end of the incubation period for a week, the spheroids were collected, reseeded and treated with α‐Mangostin for another week to obtain secondary spheroids. Further, secondary spheroids were collected, reseeded and treated with α‐Mangostin for another week to obtain tertiary spheroids. Cell viability in the spheroids was measured by trypan blue assay at the end of 7, 14 and 21 d. Data represent mean ± SD. *, &, #, %, ˄ and ** = significantly different from control, P < 0.05. C, In soft agar, human pancreatic CSCs were seeded and treated with α‐Mangostin (0‐10 μmol/L) for 21 d. At the end of the incubation period, the number of colonies was counted. *, #, % and @ = significantly different from control (n = 4), P < 0.05. D, Human pancreatic CSCs were treated with α‐Mangostin (0‐10 μmol/L) for 48 h. The expression of CD24, CD44 and CD133 was measured by the Western blot analysis. E, human pancreatic CSCs were treated with α‐Mangostin (0‐10 μmol/L) for 48 h, and the expression of Nanog, Oct4, Sox2, KLF4 and c‐Myc was measured by the Western blot analysis. β‐actin was used as a loading control

    Techniques Used: Expressing, Isolation, Incubation, Western Blot

    anti cd24  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Cell Signaling Technology Inc anti cd24
    Anti Cd24, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti cd24/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti cd24 - by Bioz Stars, 2023-02
    94/100 stars

    Images

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94
    Cell Signaling Technology Inc anti cd24
    3-Cl-AHPC-mediated inhibition and degradation of pancreatic cancer stem cells spheres of CD133 + , CD44 + <t>/CD24</t> + /EpCAM + , and stem-like spheres of CD44 + <t>/CD24</t> + PANC-1 cells. ((a), (b) and (c), (d)) 3-Cl-AHPC and AHP3 exposure resulted in inhibition of CD44 + /CD24 + /EpCAM + and CD133 + cells growth and sphere formation and degradation of the derived spheres. ((e), (f)) AHP3 and 3-Cl-AHPC inhibited sphere formation and inhibition of growth and degradation of the CD44 + /CD24 + -derived spheres. For sphere formation, the CD44 + /CD24 + /EpCAM + , CD133 + , and CD44 + /CD24 + cells were sorted by flow cytometry and approximately 200–300 cells were seeded with B27 containing DMEM/F12 medium in 96-well low attachment plates and 1 μ M 3-Cl-AHPC or AHP3 added either the day after seeding or 7 days following sphere formation. The sizes of spheres were photographed and measured on a 100 μ m scale and magnification 400X using Olympus fluorescence microscope digital camera software and DP2-BSW software. The error bars represent the mean of 15 sphere determinations ± the standard deviation. ** was significantly different in comparison to control spheres. Data were analyzed by ANOVA, Tukey HSD test for multiple comparisons. ** P < 0.01 versus control.
    Anti Cd24, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti cd24/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti cd24 - by Bioz Stars, 2023-02
    94/100 stars
      Buy from Supplier

    94
    Cell Signaling Technology Inc mouse monoclonal anti cd24
    3-Cl-AHPC-mediated inhibition and degradation of pancreatic cancer stem cells spheres of CD133 + , CD44 + <t>/CD24</t> + /EpCAM + , and stem-like spheres of CD44 + <t>/CD24</t> + PANC-1 cells. ((a), (b) and (c), (d)) 3-Cl-AHPC and AHP3 exposure resulted in inhibition of CD44 + /CD24 + /EpCAM + and CD133 + cells growth and sphere formation and degradation of the derived spheres. ((e), (f)) AHP3 and 3-Cl-AHPC inhibited sphere formation and inhibition of growth and degradation of the CD44 + /CD24 + -derived spheres. For sphere formation, the CD44 + /CD24 + /EpCAM + , CD133 + , and CD44 + /CD24 + cells were sorted by flow cytometry and approximately 200–300 cells were seeded with B27 containing DMEM/F12 medium in 96-well low attachment plates and 1 μ M 3-Cl-AHPC or AHP3 added either the day after seeding or 7 days following sphere formation. The sizes of spheres were photographed and measured on a 100 μ m scale and magnification 400X using Olympus fluorescence microscope digital camera software and DP2-BSW software. The error bars represent the mean of 15 sphere determinations ± the standard deviation. ** was significantly different in comparison to control spheres. Data were analyzed by ANOVA, Tukey HSD test for multiple comparisons. ** P < 0.01 versus control.
    Mouse Monoclonal Anti Cd24, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse monoclonal anti cd24/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse monoclonal anti cd24 - by Bioz Stars, 2023-02
    94/100 stars
      Buy from Supplier

    94
    Cell Signaling Technology Inc cd24
    3-Cl-AHPC-mediated inhibition and degradation of pancreatic cancer stem cells spheres of CD133 + , CD44 + <t>/CD24</t> + /EpCAM + , and stem-like spheres of CD44 + <t>/CD24</t> + PANC-1 cells. ((a), (b) and (c), (d)) 3-Cl-AHPC and AHP3 exposure resulted in inhibition of CD44 + /CD24 + /EpCAM + and CD133 + cells growth and sphere formation and degradation of the derived spheres. ((e), (f)) AHP3 and 3-Cl-AHPC inhibited sphere formation and inhibition of growth and degradation of the CD44 + /CD24 + -derived spheres. For sphere formation, the CD44 + /CD24 + /EpCAM + , CD133 + , and CD44 + /CD24 + cells were sorted by flow cytometry and approximately 200–300 cells were seeded with B27 containing DMEM/F12 medium in 96-well low attachment plates and 1 μ M 3-Cl-AHPC or AHP3 added either the day after seeding or 7 days following sphere formation. The sizes of spheres were photographed and measured on a 100 μ m scale and magnification 400X using Olympus fluorescence microscope digital camera software and DP2-BSW software. The error bars represent the mean of 15 sphere determinations ± the standard deviation. ** was significantly different in comparison to control spheres. Data were analyzed by ANOVA, Tukey HSD test for multiple comparisons. ** P < 0.01 versus control.
    Cd24, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cd24/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cd24 - by Bioz Stars, 2023-02
    94/100 stars
      Buy from Supplier

    Image Search Results


    3-Cl-AHPC-mediated inhibition and degradation of pancreatic cancer stem cells spheres of CD133 + , CD44 + /CD24 + /EpCAM + , and stem-like spheres of CD44 + /CD24 + PANC-1 cells. ((a), (b) and (c), (d)) 3-Cl-AHPC and AHP3 exposure resulted in inhibition of CD44 + /CD24 + /EpCAM + and CD133 + cells growth and sphere formation and degradation of the derived spheres. ((e), (f)) AHP3 and 3-Cl-AHPC inhibited sphere formation and inhibition of growth and degradation of the CD44 + /CD24 + -derived spheres. For sphere formation, the CD44 + /CD24 + /EpCAM + , CD133 + , and CD44 + /CD24 + cells were sorted by flow cytometry and approximately 200–300 cells were seeded with B27 containing DMEM/F12 medium in 96-well low attachment plates and 1 μ M 3-Cl-AHPC or AHP3 added either the day after seeding or 7 days following sphere formation. The sizes of spheres were photographed and measured on a 100 μ m scale and magnification 400X using Olympus fluorescence microscope digital camera software and DP2-BSW software. The error bars represent the mean of 15 sphere determinations ± the standard deviation. ** was significantly different in comparison to control spheres. Data were analyzed by ANOVA, Tukey HSD test for multiple comparisons. ** P < 0.01 versus control.

    Journal: Journal of Oncology

    Article Title: Adamantyl Retinoid-Related Molecules Induce Apoptosis in Pancreatic Cancer Cells by Inhibiting IGF-1R and Wnt/ β -Catenin Pathways

    doi: 10.1155/2012/796729

    Figure Lengend Snippet: 3-Cl-AHPC-mediated inhibition and degradation of pancreatic cancer stem cells spheres of CD133 + , CD44 + /CD24 + /EpCAM + , and stem-like spheres of CD44 + /CD24 + PANC-1 cells. ((a), (b) and (c), (d)) 3-Cl-AHPC and AHP3 exposure resulted in inhibition of CD44 + /CD24 + /EpCAM + and CD133 + cells growth and sphere formation and degradation of the derived spheres. ((e), (f)) AHP3 and 3-Cl-AHPC inhibited sphere formation and inhibition of growth and degradation of the CD44 + /CD24 + -derived spheres. For sphere formation, the CD44 + /CD24 + /EpCAM + , CD133 + , and CD44 + /CD24 + cells were sorted by flow cytometry and approximately 200–300 cells were seeded with B27 containing DMEM/F12 medium in 96-well low attachment plates and 1 μ M 3-Cl-AHPC or AHP3 added either the day after seeding or 7 days following sphere formation. The sizes of spheres were photographed and measured on a 100 μ m scale and magnification 400X using Olympus fluorescence microscope digital camera software and DP2-BSW software. The error bars represent the mean of 15 sphere determinations ± the standard deviation. ** was significantly different in comparison to control spheres. Data were analyzed by ANOVA, Tukey HSD test for multiple comparisons. ** P < 0.01 versus control.

    Article Snippet: Anti-CD44, anti-IGF-1R β , anti- β -catenin, anti-caspase 3, anti-cleaved caspase-3, and activated anti-cleaved Notch-1 (Val 1744) were from Cell Signaling Technology (Boston, MA); anti-CD24 and anti-cyclin D1 antibodies from Santa Cruz Biotechnology (Santa Cruz, CA); CD133-PE and CD326-FITC (EpCAM) from Miltenyi Biotec Inc. (Auburn, CA), and anti- α -tubulin antibody from Oncogene Research Products (Boston, MA).

    Techniques: Inhibition, Derivative Assay, Flow Cytometry, Fluorescence, Microscopy, Software, Standard Deviation

    3-Cl-AHPC- (1 μ M) and AHP3- (1 μ M) mediated inhibition of CD44 + /CD24 + stem-like cell sphere formation and degradation of spheres derived from MiaPaCa-2 and Capan-2 cell lines. 3-Cl-AHPC and AHP3 were added at the time cells were seeded ((a), (c)) or 7 days after cells sphere formation ((b), (d)). The ARR affect on sphere growth was assessed at days 7 and 14 (a), days 7 and 14 (b), days 14 and 21 (c), and days 14 and 21 (d), respectively. The error bars represent the mean of 15 sphere determinations ± the standard deviation. ** Was significantly different in comparison to control spheres. Data were analyzed by ANOVA, Tukey HSD test for multiple comparisons. ** P < 0.01 versus control.

    Journal: Journal of Oncology

    Article Title: Adamantyl Retinoid-Related Molecules Induce Apoptosis in Pancreatic Cancer Cells by Inhibiting IGF-1R and Wnt/ β -Catenin Pathways

    doi: 10.1155/2012/796729

    Figure Lengend Snippet: 3-Cl-AHPC- (1 μ M) and AHP3- (1 μ M) mediated inhibition of CD44 + /CD24 + stem-like cell sphere formation and degradation of spheres derived from MiaPaCa-2 and Capan-2 cell lines. 3-Cl-AHPC and AHP3 were added at the time cells were seeded ((a), (c)) or 7 days after cells sphere formation ((b), (d)). The ARR affect on sphere growth was assessed at days 7 and 14 (a), days 7 and 14 (b), days 14 and 21 (c), and days 14 and 21 (d), respectively. The error bars represent the mean of 15 sphere determinations ± the standard deviation. ** Was significantly different in comparison to control spheres. Data were analyzed by ANOVA, Tukey HSD test for multiple comparisons. ** P < 0.01 versus control.

    Article Snippet: Anti-CD44, anti-IGF-1R β , anti- β -catenin, anti-caspase 3, anti-cleaved caspase-3, and activated anti-cleaved Notch-1 (Val 1744) were from Cell Signaling Technology (Boston, MA); anti-CD24 and anti-cyclin D1 antibodies from Santa Cruz Biotechnology (Santa Cruz, CA); CD133-PE and CD326-FITC (EpCAM) from Miltenyi Biotec Inc. (Auburn, CA), and anti- α -tubulin antibody from Oncogene Research Products (Boston, MA).

    Techniques: Inhibition, Derivative Assay, Standard Deviation

    Dose-response effect of 3-Cl-AHPC on CD44 + /CD24 + cells sphere formation and apoptosis in PANC-1 sphere cells. (a) Addition of 0.25, 0.5, and 1.0 μ M 3-Cl-AHPC added at time of cell seeding inhibited sphere formation at 7 and 14 days. (b) 0.5 and 1.0 μ M 3-Cl-AHPC inhibited sphere formation when added 7 days following sphere formation. (c) 1.0 μ M 3-Cl-AHPC induced apoptosis in CD44 + /CD24 + sphere cells as indicated by nuclear fragmentation detected by acridine orange/ethidium bromide and (d) DAPI staining. Spheres were visualized and photographed utilizing a fluorescence microscope. (e) Apoptosis of sphere cells as demonstrated by TUNEL assay. CD44 + /CD24 + spheres were treated with 1.0 μ M ARRs for 7 days (7D) after sphere formation. Details of slides preparation, visualization, antibodies utilized, and TUNNEL assay methodologies were as described in .

    Journal: Journal of Oncology

    Article Title: Adamantyl Retinoid-Related Molecules Induce Apoptosis in Pancreatic Cancer Cells by Inhibiting IGF-1R and Wnt/ β -Catenin Pathways

    doi: 10.1155/2012/796729

    Figure Lengend Snippet: Dose-response effect of 3-Cl-AHPC on CD44 + /CD24 + cells sphere formation and apoptosis in PANC-1 sphere cells. (a) Addition of 0.25, 0.5, and 1.0 μ M 3-Cl-AHPC added at time of cell seeding inhibited sphere formation at 7 and 14 days. (b) 0.5 and 1.0 μ M 3-Cl-AHPC inhibited sphere formation when added 7 days following sphere formation. (c) 1.0 μ M 3-Cl-AHPC induced apoptosis in CD44 + /CD24 + sphere cells as indicated by nuclear fragmentation detected by acridine orange/ethidium bromide and (d) DAPI staining. Spheres were visualized and photographed utilizing a fluorescence microscope. (e) Apoptosis of sphere cells as demonstrated by TUNEL assay. CD44 + /CD24 + spheres were treated with 1.0 μ M ARRs for 7 days (7D) after sphere formation. Details of slides preparation, visualization, antibodies utilized, and TUNNEL assay methodologies were as described in .

    Article Snippet: Anti-CD44, anti-IGF-1R β , anti- β -catenin, anti-caspase 3, anti-cleaved caspase-3, and activated anti-cleaved Notch-1 (Val 1744) were from Cell Signaling Technology (Boston, MA); anti-CD24 and anti-cyclin D1 antibodies from Santa Cruz Biotechnology (Santa Cruz, CA); CD133-PE and CD326-FITC (EpCAM) from Miltenyi Biotec Inc. (Auburn, CA), and anti- α -tubulin antibody from Oncogene Research Products (Boston, MA).

    Techniques: Staining, Fluorescence, Microscopy, TUNEL Assay

    3-Cl-AHPC and AHP induced apoptosis in PANC-1 CD44 + /CD24 + cells and 3-Cl-AHPC decreased expression of IGF-1R, cyclin D1, and β -catenin in pancreatic cancer cells. (a) Percentage of total apoptotic cells. (b) Percentage of CD44 + /CD24 + cells in the early (Annexin V-FITC positive and PI negative) or late (Annexin V-FITC positive and PI positive) apoptotic cell populations. (c) Percentage of total CD44 + /CD24 + apoptotic cells (Annexin V-FITC positive and PI positive). Cells were treated with 1.0 μ M 3-Cl-AHPC and AHP3 for 96 h. Antibody-conjugated markers CD44-APC-Cy7, CD24-APC, Annexin V-FITC, and PI were used to detect apoptotic and CD44 + /CD24 + cells from the same samples. The error bars represent the mean of three separate determinations ± the standard deviation. ((d), (e)) IGF-1R, cyclin D1, and β -catenin expression decreased following 3-Cl-AHPC exposure in pancreatic cancer cells.

    Journal: Journal of Oncology

    Article Title: Adamantyl Retinoid-Related Molecules Induce Apoptosis in Pancreatic Cancer Cells by Inhibiting IGF-1R and Wnt/ β -Catenin Pathways

    doi: 10.1155/2012/796729

    Figure Lengend Snippet: 3-Cl-AHPC and AHP induced apoptosis in PANC-1 CD44 + /CD24 + cells and 3-Cl-AHPC decreased expression of IGF-1R, cyclin D1, and β -catenin in pancreatic cancer cells. (a) Percentage of total apoptotic cells. (b) Percentage of CD44 + /CD24 + cells in the early (Annexin V-FITC positive and PI negative) or late (Annexin V-FITC positive and PI positive) apoptotic cell populations. (c) Percentage of total CD44 + /CD24 + apoptotic cells (Annexin V-FITC positive and PI positive). Cells were treated with 1.0 μ M 3-Cl-AHPC and AHP3 for 96 h. Antibody-conjugated markers CD44-APC-Cy7, CD24-APC, Annexin V-FITC, and PI were used to detect apoptotic and CD44 + /CD24 + cells from the same samples. The error bars represent the mean of three separate determinations ± the standard deviation. ((d), (e)) IGF-1R, cyclin D1, and β -catenin expression decreased following 3-Cl-AHPC exposure in pancreatic cancer cells.

    Article Snippet: Anti-CD44, anti-IGF-1R β , anti- β -catenin, anti-caspase 3, anti-cleaved caspase-3, and activated anti-cleaved Notch-1 (Val 1744) were from Cell Signaling Technology (Boston, MA); anti-CD24 and anti-cyclin D1 antibodies from Santa Cruz Biotechnology (Santa Cruz, CA); CD133-PE and CD326-FITC (EpCAM) from Miltenyi Biotec Inc. (Auburn, CA), and anti- α -tubulin antibody from Oncogene Research Products (Boston, MA).

    Techniques: Expressing, Standard Deviation

    3-Cl-AHPC decreased expression of IGF-1R, cyclin D1, β -catenin, and cleaved Notch-1 and increased levels of cleaved-caspase-3 in CD44 + /CD24 + spheres. (a) IGF-1R, cyclin D1, and β -catenin expression decreased cleaved-caspase-3 increased with no change in Notch-1 protein levels in spheres following exposure to 3-Cl-AHPC. Pancreatic cancer cells and PANC-1 spheres were exposed to 1.0 μ M 3-Cl-AHPC for 7 days. Western blots were prepared as described in Materials and Methods. (b) mRNA expression of GLI1, GLI2, and Ptch1 in PANC-1 cells. Cells were grown in the presence of 1 μ M 3-Cl-AHPC or vehicle alone (control). (c) Knockdown of IGF-1R expression by sh-IGF-1R inhibited sphere formation and enhanced ARR inhibition of sphere formation. The error bars represent the mean of three separate determinations +/− the standard deviation. •• was significantly different between spheres comprised of sh-vector cells treated with vehicle and 3-Cl-AHPC or AHP3. ♦♦ was significantly different between spheres comprised of sh-vector and IGF-1R-KD1 or IGF-1R-KD 2 at 7 and 14 days, respectively. ** was significantly different in comparison between IGF-1R-KDl/IGF-1R-KD2 spheres (vehicle treated) and IGF-1R-KD1/IGF-1R-KD2 spheres treated with 3-Cl-AHPC or AHP3. Data were analyzed by ANOVA, Tukey HSD test for multiple comparisons, ♦♦, ••, and ** P < 0.01. (d) Knockdown (KD) of IGF-1R enhanced AHP3- and 3-Cl-AHPC-mediated apoptosis in the PANC-1 cells and IGF-IR protein expression in IGF-1R knockdown cells. Apoptosis was assessed using acridine orange/ethidium bromide staining as described in .

    Journal: Journal of Oncology

    Article Title: Adamantyl Retinoid-Related Molecules Induce Apoptosis in Pancreatic Cancer Cells by Inhibiting IGF-1R and Wnt/ β -Catenin Pathways

    doi: 10.1155/2012/796729

    Figure Lengend Snippet: 3-Cl-AHPC decreased expression of IGF-1R, cyclin D1, β -catenin, and cleaved Notch-1 and increased levels of cleaved-caspase-3 in CD44 + /CD24 + spheres. (a) IGF-1R, cyclin D1, and β -catenin expression decreased cleaved-caspase-3 increased with no change in Notch-1 protein levels in spheres following exposure to 3-Cl-AHPC. Pancreatic cancer cells and PANC-1 spheres were exposed to 1.0 μ M 3-Cl-AHPC for 7 days. Western blots were prepared as described in Materials and Methods. (b) mRNA expression of GLI1, GLI2, and Ptch1 in PANC-1 cells. Cells were grown in the presence of 1 μ M 3-Cl-AHPC or vehicle alone (control). (c) Knockdown of IGF-1R expression by sh-IGF-1R inhibited sphere formation and enhanced ARR inhibition of sphere formation. The error bars represent the mean of three separate determinations +/− the standard deviation. •• was significantly different between spheres comprised of sh-vector cells treated with vehicle and 3-Cl-AHPC or AHP3. ♦♦ was significantly different between spheres comprised of sh-vector and IGF-1R-KD1 or IGF-1R-KD 2 at 7 and 14 days, respectively. ** was significantly different in comparison between IGF-1R-KDl/IGF-1R-KD2 spheres (vehicle treated) and IGF-1R-KD1/IGF-1R-KD2 spheres treated with 3-Cl-AHPC or AHP3. Data were analyzed by ANOVA, Tukey HSD test for multiple comparisons, ♦♦, ••, and ** P < 0.01. (d) Knockdown (KD) of IGF-1R enhanced AHP3- and 3-Cl-AHPC-mediated apoptosis in the PANC-1 cells and IGF-IR protein expression in IGF-1R knockdown cells. Apoptosis was assessed using acridine orange/ethidium bromide staining as described in .

    Article Snippet: Anti-CD44, anti-IGF-1R β , anti- β -catenin, anti-caspase 3, anti-cleaved caspase-3, and activated anti-cleaved Notch-1 (Val 1744) were from Cell Signaling Technology (Boston, MA); anti-CD24 and anti-cyclin D1 antibodies from Santa Cruz Biotechnology (Santa Cruz, CA); CD133-PE and CD326-FITC (EpCAM) from Miltenyi Biotec Inc. (Auburn, CA), and anti- α -tubulin antibody from Oncogene Research Products (Boston, MA).

    Techniques: Expressing, Western Blot, Inhibition, Standard Deviation, Plasmid Preparation, Staining

    3-Cl-AHPC mediated inhibition of the activation of TCF/LEF in Wnt/ β -catenin pathway and decreased of β -catenin nuclear localization. (a) 3-Cl-AHPC decreased nuclear β -catenin as indicated by Western blot using nuclear extracts and densitometric quantification. (b) Nuclear β -catenin in control- (i) and 3-Cl-AHPC- (ii) treated PANC-1 cells using confocal fluorescent microscope (magnification 40X). Cells were grown in eight chambered slides and then treated with 3-Cl-AHPC for 24 h. Slide was prepared as described in . DAPI was used for nuclear staining for 1 min and mounted the slide with prolong gold antifade kit. (c) 3-Cl-AHPC inhibited TCF/LEF activity in Wnt/ β -catenin signaling in stably transfected Cignal TCF/LEF-Luc reporter PANC-1 cell lines and 50 mM LiCl was used as a positive control. For CD44/CD24 cells, TCF/LEF stably transfected cells were sorted by flow cytometry and followed the procedure same as wild type (Wt) stable cell line. Luciferase promoter activity values are expressed as fold using a total protein concentration for internal normalization. The error bars represent the mean of three separate determinations ± the standard deviation (SD). (d) 3-Cl-AHPC decreased Wnt/ β -catenin signaling responsive c-Myc protein. ((e) and (f)) Knock down of β -catenin inhibited cell proliferations and enhanced more apoptosis in sh- β -catenin knockdown (KD) PANC-1 cell lines. Proliferation inhibition was evaluated after 72 h of seeding the cells by MTT assay and expressed as absorbance measured at 570 nm. The error bars represent the mean of three separate determinations ± the standard deviation (SD). ** (<0.01) was significantly different in comparison between sh-vector and Catenin-KDl/Catenin-KD2 and also between sh-vector and catenin-KD1/Catenin-KD2 treated with 3-Cl-AHPC, respectively.

    Journal: Journal of Oncology

    Article Title: Adamantyl Retinoid-Related Molecules Induce Apoptosis in Pancreatic Cancer Cells by Inhibiting IGF-1R and Wnt/ β -Catenin Pathways

    doi: 10.1155/2012/796729

    Figure Lengend Snippet: 3-Cl-AHPC mediated inhibition of the activation of TCF/LEF in Wnt/ β -catenin pathway and decreased of β -catenin nuclear localization. (a) 3-Cl-AHPC decreased nuclear β -catenin as indicated by Western blot using nuclear extracts and densitometric quantification. (b) Nuclear β -catenin in control- (i) and 3-Cl-AHPC- (ii) treated PANC-1 cells using confocal fluorescent microscope (magnification 40X). Cells were grown in eight chambered slides and then treated with 3-Cl-AHPC for 24 h. Slide was prepared as described in . DAPI was used for nuclear staining for 1 min and mounted the slide with prolong gold antifade kit. (c) 3-Cl-AHPC inhibited TCF/LEF activity in Wnt/ β -catenin signaling in stably transfected Cignal TCF/LEF-Luc reporter PANC-1 cell lines and 50 mM LiCl was used as a positive control. For CD44/CD24 cells, TCF/LEF stably transfected cells were sorted by flow cytometry and followed the procedure same as wild type (Wt) stable cell line. Luciferase promoter activity values are expressed as fold using a total protein concentration for internal normalization. The error bars represent the mean of three separate determinations ± the standard deviation (SD). (d) 3-Cl-AHPC decreased Wnt/ β -catenin signaling responsive c-Myc protein. ((e) and (f)) Knock down of β -catenin inhibited cell proliferations and enhanced more apoptosis in sh- β -catenin knockdown (KD) PANC-1 cell lines. Proliferation inhibition was evaluated after 72 h of seeding the cells by MTT assay and expressed as absorbance measured at 570 nm. The error bars represent the mean of three separate determinations ± the standard deviation (SD). ** (<0.01) was significantly different in comparison between sh-vector and Catenin-KDl/Catenin-KD2 and also between sh-vector and catenin-KD1/Catenin-KD2 treated with 3-Cl-AHPC, respectively.

    Article Snippet: Anti-CD44, anti-IGF-1R β , anti- β -catenin, anti-caspase 3, anti-cleaved caspase-3, and activated anti-cleaved Notch-1 (Val 1744) were from Cell Signaling Technology (Boston, MA); anti-CD24 and anti-cyclin D1 antibodies from Santa Cruz Biotechnology (Santa Cruz, CA); CD133-PE and CD326-FITC (EpCAM) from Miltenyi Biotec Inc. (Auburn, CA), and anti- α -tubulin antibody from Oncogene Research Products (Boston, MA).

    Techniques: Inhibition, Activation Assay, Western Blot, Microscopy, Staining, Activity Assay, Stable Transfection, Transfection, Positive Control, Flow Cytometry, Luciferase, Protein Concentration, Standard Deviation, MTT Assay, Plasmid Preparation