hct116 cells (ATCC)
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hct116 cells
Hct116 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 4089 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/representative+tissue+microarray+staining/HCT+116/pmc06663864-387-12-27
Average 99 stars, based on 4089 article reviews
Hct116 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 4089 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/representative+tissue+microarray+staining/HCT+116/pmc06663864-387-12-27
Average 99 stars, based on 4089 article reviews
hct116 cells - by Bioz Stars,
2026-10
99/100 stars
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1) Product Images from "Trophoblast cell surface antigen 2 (Trop-2) phosphorylation by protein kinase C α/δ (PKCα/δ) enhances cell motility"
Article Title: Trophoblast cell surface antigen 2 (Trop-2) phosphorylation by protein kinase C α/δ (PKCα/δ) enhances cell motility
Journal: The Journal of Biological Chemistry
doi: 10.1074/jbc.RA119.008084
Figure Legend Snippet: Phosphorylation of Trop-2. A, FLAG-tagged Trop-2 was immunoprecipitated (IP) from lysates of HCT116/M (M) and HCT116/WT (WT: #1) cells and subjected to SDS-PAGE, followed by immunoblotting (IB). B, FLAG-tagged Trop-2 immunoprecipitated from a lysate of HCT116/WT (#1) cells with anti-FLAG magnetic beads was treated with or without N-glycanase and then subjected to SDS-PAGE, followed by immunoblotting. C, after N-glycanase treatment, FLAG-tagged Trop-2 obtained from a lysate of HCT116/WT (#1) cells as described above was subjected to SDS-PAGE using a Phos-tag–containing gel (Phos-tag SDS-PAGE), followed by immunoblotting. The top and bottom bands represent phosphorylated and unphosphorylated FLAG-tagged Trop-2, respectively. D, schematic model of WT and mutated FLAG-tagged Trop-2. ED, ectodomain; TM, transmembrane domain; CD, cytoplasmic domain. E, after N-glycanase treatment, FLAG-tagged Trop-2 immunoprecipitated from lysates of HCT116/WT (#1 and #2), HCT116/S303A (S303A: #1 and #2), and HCT116/S322A (S322A: #1 and #2) cells as described above was subjected to Phos-tag (top) and ordinary (bottom) SDS-PAGE, followed by immunoblotting. The arrowhead indicates phosphorylated FLAG-tagged Trop-2. F, intensities of the bands in the Phos-tag gel panel in E were measured, and then the ratio of phosphorylated to total FLAG-tagged Trop-2 was calculated (means ± S.E. (error bars), n = 4).
Techniques Used: Phospho-proteomics, Immunoprecipitation, SDS Page, Western Blot, Magnetic Beads
Figure Legend Snippet: Interaction of Trop-2 with claudin-7. A, lysates (left) of HCT116/M (M) and HCT116/WT (WT: #1) cells and the immunoprecipitates (IP) (right) obtained from the lysates with anti-FLAG magnetic beads were subjected to SDS-PAGE, followed by immunoblotting (IB), and detection of claudin-7, occludin, E-cadherin, and FLAG-tagged Trop-2. β-Actin served as a loading control. B and C, HCT116 cells were stably transfected with both a Tet-ON regulator plasmid and a response plasmid–inserted FLAG-tagged Trop-2 gene. Lysates of the cells treated with or without 10 μg/ml doxycycline (Dox) for 1–6 days were subjected to SDS-PAGE, followed by immunoblotting. β-Actin served as a loading control. The intensity of the claudin-7 and β-actin bands in B was measured, and the ratio of claudin-7 to β-actin in each Dox-treated cell is indicated, with that in Dox-nontreated cells being taken as 1 (means ± S.E. (error bars), n = 3; *, p < 0.05; **, p < 0.01). D and E, immunoprecipitates from the lysates of HCT116/WT (#1), HCT116/S322A (SA: #1), and HCT116/S322E (SE: #1) cells with anti-claudin-7 antibodies were subjected to SDS-PAGE, followed by immunoblotting. Histograms show the intensity of the FLAG-tagged Trop-2 bands, each density being normalized as to that of claudin-7, and the value for HCT116/WT (#1) cells was taken as 1 (means ± S.E., n = 3; *, p < 0.05; **, p < 0.01). F, after treatment of HCT116/WT (#1) cells with PMA or 4α-PMA for 2 h, their lysates (left) and claudin-7 coimmunoprecipitated with anti-FLAG magnetic beads (right) were subjected to SDS-PAGE, followed by immunoblotting, and detection with anti-claudin-7 and anti-FLAG antibodies. β-Actin served as a loading control.
Techniques Used: Magnetic Beads, SDS Page, Western Blot, Control, Stable Transfection, Transfection, Plasmid Preparation
Figure Legend Snippet: Expression and distribution of Trop-2 and claudin-7 in various types of Trop-2–expressing cells and in cancer tissues. A, levels of claudin-7 mRNA in HCT116/M (M), HCT116/WT (WT: #1), HCT116/S322A (SA: #1), and HCT116/S322E (SE: #1) cells were determined by DNA microarray analysis, and its level in HCT116/M cells was taken as 1. B, expression of claudin-7 protein in the four cell types described above was analyzed by SDS-PAGE, followed by immunoblotting, and that in HCT116/M cells was normalized as to β-actin, and the value for HCT116/M cells was taken as 1. β-Actin served as a loading control (means ± S.E. (error bars), n = 4; *, p < 0.05; **, p < 0.01). C, distribution of claudin-7 (green) and FLAG-tagged Trop-2 (magenta) in HCT116/M, HCT116/WT (#1), HCT116/S322A (#1), and HCT116/S322E (#1) cells was observed immunochemically. Nuclei were stained with DAPI (blue). Scale bars, 10 μm. D, distribution of claudin-7 and Trop-2 was observed immunohistochemically in colorectal cancer tissues. Nuclei were stained with hematoxylin. Scale bars, 100 μm.
Techniques Used: Expressing, Microarray, SDS Page, Western Blot, Control, Staining
Figure Legend Snippet: Phosphorylation of Trop-2 by PKCα and PKCδ. A, expression of PKC and PKD isoforms in HCT116/WT (WT: #1) cells was examined by DNA microarray analysis. B and C, HCT116/WT (#1) cells were treated with or without a PKC or PKD inhibitor for 1 h and subsequently with PMA or 4α-PMA for 2 h. Each cell lysate was subjected to SDS-PAGE, followed by immunoblotting. The intensities of the bands in B were determined. Histograms show the inhibitory effect of each inhibitor, the levels of phosphorylated Trop-2 being represented as the ratio of phosphorylated to total FLAG-tagged Trop-2 (p-Trop-2/FLAG), and the ratio obtained for 4α-PMA–treated cells was taken as 1 (means ± S.E. (error bars), n = 3; **, p < 0.01; NS, not significant). D and E, HCT116/WT (#1) cells were transiently transfected with PKCα siRNA (#1), PKCδ siRNA (#1), both PKCα and PKCδ siRNAs (#1), or a control siRNA and then treated with PMA or 4α-PMA for 2 h. A lysate of each cell type was subjected to SDS-PAGE, followed by immunoblotting. β-Actin served as a loading control. The intensities of the bands in D were determined, and the relative value (p-Trop-2/FLAG) was calculated as described in B and C. Histograms show the p-Trop-2/FLAG ratio in PMA-treated cells and that in 4α-PMA–treated cells (mean ± S.E., n = 3; **, p < 0.01; NS, not significant).
Techniques Used: Phospho-proteomics, Expressing, Microarray, SDS Page, Western Blot, Transfection, Control
Figure Legend Snippet: Effects of Trop-2 phosphorylation on motility of WT and mutated Trop-2–expressing HCT116 cells. A and B, migration of HCT116/M (M, n = 7), HCT116/WT (WT: #1, n = 4), HCT116/S322A (SA: #1, n = 4), and HCT116/S322E (SE: #1, n = 3) cells was evaluated by means of Transwell assays. A representative photograph of each migrated cell type is shown in A. Scale bars, 100 μm. The number of migrated cells per field was determined and is shown as a histogram in B (means ± S.E. (error bars); **, p < 0.01). C, proliferation of HCT116/M (n = 7), HCT116/WT (#1: n = 4), HCT116/S322A (#1: n = 4), and HCT116/S322E (#1: n = 3) cells was analyzed using MTT assays (means ± S.E.). D and E, migration of HCT116/WT (#1) cells treated with or without PKC inhibitors was measured as described in A and B. A representative photograph of each migrated cell is shown in D. Scale bars, 100 μm. The number of migrated cells per field was determined as described in A and B and is shown as a histogram in E (means ± S.E., n = 3; **, p < 0.01; NS, not significant). F, Proliferation of cells was examined using MTT assays as described in C (means ± S.E., n = 3). G and H, migration of HCT116/WT (#1) cells treated with both PKCα and PKCδ siRNAs (#1) or a control siRNA and then with PMA or 4α-PMA was analyzed by using Transwells. A representative photograph of each migrated cell type is shown in G. Scale bars, 100 μm. The number of migrated cells per field was determined as described in A and B and is shown as a histogram in H (means ± S.E., n = 3; **, p < 0.01; NS, not significant). I, proliferation of cells was analyzed using MTT assays as described in C (means ± S.E., n = 3).
Techniques Used: Phospho-proteomics, Expressing, Migration, Control
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