er tracker green  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc er tracker green
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    Cell Signaling Technology Inc ikkβ
    Wee1/CDC2 and nuclear factor κB (NF-κB) signaling pathway molecules in parental tumor cells had heterogeneous expression levels and were upregulated by cisplatin (CDDP). Tumor cells were seeded in a 10-cm cell culture dish (1×10 5 cells/dish) and cultured for 48 hours. Cells were treated with CDDP (25 μg/mL) in fresh medium for 6 hours when they reached 70%–80% confluence and then harvested. Parental cells were treated with fresh medium alone. Protein quantification was performed after cell lysis, and 30 μg protein per well was subjected to western blot analysis. The protein expression levels of Wee1/CDC2 and NF-κB pathway <t>molecules</t> <t>IKKα,</t> <t>IKKβ,</t> RelA, and IκB in conventional cells were significantly different, with the hierarchy Saos2 > U2OS > MG63 for OS cell lines (p < 0.01) (A), UN-SCC46 > UM-SCC47 > UM-SCC1 for head and neck squamous cell lines (p < 0.01) (B), and PDX-OS3 > PDX-OS2 > PDX-OS1 for PDX OS cell lines (p < 0.01) (C). CDDP significantly increased the protein expression levels of Wee1, IKKα, IKKβ, and RelA and decreased that of IκB in these tumor cells (p < 0.01).
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    1) Product Images from "Enrichment of Wee1/CDC2 and NF-κB Signaling Pathway Constituents Mutually Contributes to CDDP Resistance in Human Osteosarcoma"

    Article Title: Enrichment of Wee1/CDC2 and NF-κB Signaling Pathway Constituents Mutually Contributes to CDDP Resistance in Human Osteosarcoma

    Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

    doi: 10.4143/crt.2021.320

    Wee1/CDC2 and nuclear factor κB (NF-κB) signaling pathway molecules in parental tumor cells had heterogeneous expression levels and were upregulated by cisplatin (CDDP). Tumor cells were seeded in a 10-cm cell culture dish (1×10 5 cells/dish) and cultured for 48 hours. Cells were treated with CDDP (25 μg/mL) in fresh medium for 6 hours when they reached 70%–80% confluence and then harvested. Parental cells were treated with fresh medium alone. Protein quantification was performed after cell lysis, and 30 μg protein per well was subjected to western blot analysis. The protein expression levels of Wee1/CDC2 and NF-κB pathway molecules IKKα, IKKβ, RelA, and IκB in conventional cells were significantly different, with the hierarchy Saos2 > U2OS > MG63 for OS cell lines (p < 0.01) (A), UN-SCC46 > UM-SCC47 > UM-SCC1 for head and neck squamous cell lines (p < 0.01) (B), and PDX-OS3 > PDX-OS2 > PDX-OS1 for PDX OS cell lines (p < 0.01) (C). CDDP significantly increased the protein expression levels of Wee1, IKKα, IKKβ, and RelA and decreased that of IκB in these tumor cells (p < 0.01).
    Figure Legend Snippet: Wee1/CDC2 and nuclear factor κB (NF-κB) signaling pathway molecules in parental tumor cells had heterogeneous expression levels and were upregulated by cisplatin (CDDP). Tumor cells were seeded in a 10-cm cell culture dish (1×10 5 cells/dish) and cultured for 48 hours. Cells were treated with CDDP (25 μg/mL) in fresh medium for 6 hours when they reached 70%–80% confluence and then harvested. Parental cells were treated with fresh medium alone. Protein quantification was performed after cell lysis, and 30 μg protein per well was subjected to western blot analysis. The protein expression levels of Wee1/CDC2 and NF-κB pathway molecules IKKα, IKKβ, RelA, and IκB in conventional cells were significantly different, with the hierarchy Saos2 > U2OS > MG63 for OS cell lines (p < 0.01) (A), UN-SCC46 > UM-SCC47 > UM-SCC1 for head and neck squamous cell lines (p < 0.01) (B), and PDX-OS3 > PDX-OS2 > PDX-OS1 for PDX OS cell lines (p < 0.01) (C). CDDP significantly increased the protein expression levels of Wee1, IKKα, IKKβ, and RelA and decreased that of IκB in these tumor cells (p < 0.01).

    Techniques Used: Expressing, Cell Culture, Lysis, Western Blot

    er tracker  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc er tracker
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    er tracker  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc er tracker
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    Wee1/CDC2 and nuclear factor κB (NF-κB) signaling pathway molecules in parental tumor cells had heterogeneous expression levels and were upregulated by cisplatin (CDDP). Tumor cells were seeded in a 10-cm cell culture dish (1×10 5 cells/dish) and cultured for 48 hours. Cells were treated with CDDP (25 μg/mL) in fresh medium for 6 hours when they reached 70%–80% confluence and then harvested. Parental cells were treated with fresh medium alone. Protein quantification was performed after cell lysis, and 30 μg protein per well was subjected to western blot analysis. The protein expression levels of Wee1/CDC2 and NF-κB pathway <t>molecules</t> <t>IKKα,</t> <t>IKKβ,</t> RelA, and IκB in conventional cells were significantly different, with the hierarchy Saos2 > U2OS > MG63 for OS cell lines (p < 0.01) (A), UN-SCC46 > UM-SCC47 > UM-SCC1 for head and neck squamous cell lines (p < 0.01) (B), and PDX-OS3 > PDX-OS2 > PDX-OS1 for PDX OS cell lines (p < 0.01) (C). CDDP significantly increased the protein expression levels of Wee1, IKKα, IKKβ, and RelA and decreased that of IκB in these tumor cells (p < 0.01).
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    Wee1/CDC2 and nuclear factor κB (NF-κB) signaling pathway molecules in parental tumor cells had heterogeneous expression levels and were upregulated by cisplatin (CDDP). Tumor cells were seeded in a 10-cm cell culture dish (1×10 5 cells/dish) and cultured for 48 hours. Cells were treated with CDDP (25 μg/mL) in fresh medium for 6 hours when they reached 70%–80% confluence and then harvested. Parental cells were treated with fresh medium alone. Protein quantification was performed after cell lysis, and 30 μg protein per well was subjected to western blot analysis. The protein expression levels of Wee1/CDC2 and NF-κB pathway molecules IKKα, IKKβ, RelA, and IκB in conventional cells were significantly different, with the hierarchy Saos2 > U2OS > MG63 for OS cell lines (p < 0.01) (A), UN-SCC46 > UM-SCC47 > UM-SCC1 for head and neck squamous cell lines (p < 0.01) (B), and PDX-OS3 > PDX-OS2 > PDX-OS1 for PDX OS cell lines (p < 0.01) (C). CDDP significantly increased the protein expression levels of Wee1, IKKα, IKKβ, and RelA and decreased that of IκB in these tumor cells (p < 0.01).

    Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

    Article Title: Enrichment of Wee1/CDC2 and NF-κB Signaling Pathway Constituents Mutually Contributes to CDDP Resistance in Human Osteosarcoma

    doi: 10.4143/crt.2021.320

    Figure Lengend Snippet: Wee1/CDC2 and nuclear factor κB (NF-κB) signaling pathway molecules in parental tumor cells had heterogeneous expression levels and were upregulated by cisplatin (CDDP). Tumor cells were seeded in a 10-cm cell culture dish (1×10 5 cells/dish) and cultured for 48 hours. Cells were treated with CDDP (25 μg/mL) in fresh medium for 6 hours when they reached 70%–80% confluence and then harvested. Parental cells were treated with fresh medium alone. Protein quantification was performed after cell lysis, and 30 μg protein per well was subjected to western blot analysis. The protein expression levels of Wee1/CDC2 and NF-κB pathway molecules IKKα, IKKβ, RelA, and IκB in conventional cells were significantly different, with the hierarchy Saos2 > U2OS > MG63 for OS cell lines (p < 0.01) (A), UN-SCC46 > UM-SCC47 > UM-SCC1 for head and neck squamous cell lines (p < 0.01) (B), and PDX-OS3 > PDX-OS2 > PDX-OS1 for PDX OS cell lines (p < 0.01) (C). CDDP significantly increased the protein expression levels of Wee1, IKKα, IKKβ, and RelA and decreased that of IκB in these tumor cells (p < 0.01).

    Article Snippet: The primary antibodies used in this study targeted the following proteins: Wee1 (95 kD, #5285, Santa Cruz Biotechnology, Inc.), CDC2 (34 kD, #9116S, Cell Signaling Technology, Inc., Danvers, MA), pCDC2Y15 (34 kD, #8242S, Cell Signaling Technology, Inc.), IKKα (85 kD, #1930S, Cell Signaling Technology, Inc.), IKKβ (87 kD, #8943, Cell Signaling Technology, Inc.), RelA (65 kD, #8242S, Cell Signaling Technology, Inc.), pRelAS536 (65 kD, #3033S, Cell Signaling Technology, Inc.), IκB (39 kD, #9247, Cell Signaling Technology, Inc.), cyclin D1 (36 kD, #2922, Cell Signaling Technology, Inc.), Bcl-2 (26 kD, #15071, Cell Signaling Technology, Inc.), PARP (116 kD, #9532, Cell Signaling Technology, Inc.), cleaved caspase-3 (17 kD, #9662, Cell Signaling Technology, Inc.), and β-actin (42 kD, #ab8226, Abcam, Inc., MA).

    Techniques: Expressing, Cell Culture, Lysis, Western Blot