ml 1 hygromycin b u2os human osteosarcoma epithelial cells  (ATCC)


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    ATCC ml 1 hygromycin b u2os human osteosarcoma epithelial cells
    Ml 1 Hygromycin B U2os Human Osteosarcoma Epithelial Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1 hygromycin b  (InvivoGen)


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    1 hygromycin b u2os human osteosarcoma epithelial cells  (ATCC)


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    ATCC 1 hygromycin b u2os human osteosarcoma epithelial cells
    Related to Figs. and . a – c , Phosphorylation Site Preference (PSP) plots showing single-cell correlation of Rb phosphorylation at different sites in RPE-1 cells ( a ), BJ-5ta cells ( b ), or <t>U2OS</t> cells ( c ). Each phosphorylation signal was normalized by the total Rb antibody signal in the same cell and each axis was adjusted to the average phosphorylation signal in S phase of 1. Color indicates relative cell population density. A red line shows fitting with a preferential relative phosphorylation or dephosphorylation rate between the two sites (PSP coeff ) (see more details in Methods). Rb phosphorylation at T373 or T826 was plotted against S807/S811. a , b , Cells were fixed 8 h after release with growth media ( a : n = 5019, 4900 cells for T373, T826, respectively. b : n = 3409, 3502 cells for T373, T826, respectively. 1 of n = 3 biological replicates). c , Cells asynchronously cycling in growth media were fixed 6 h after DMSO or CDK4/6i (100 nM) treatment (DMSO: n = 704, 700 cells for T373, T826, respectively. CDK4/6i: n = 850, 878 cells for T373, T826, respectively. 1 of n = 3 biological replicates). d , Single-cell correlation of CDK2 activity and Rb phosphorylation at T373 and T826. Cells were fixed 48 h after release with starvation media + EGF (20 ng/mL) + CDK4/6i (1 μM). Rb phosphorylation levels as a function CDK2 activity were fitted by sigmoidal curves (sigmoidal curves with Hill coefficients. n = 1967, 2050 cells for T373, T826, respectively).
    1 Hygromycin B U2os Human Osteosarcoma Epithelial Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "An intermediate Rb–E2F activity state safeguards proliferation commitment"

    Article Title: An intermediate Rb–E2F activity state safeguards proliferation commitment

    Journal: Nature

    doi: 10.1038/s41586-024-07554-2

    Related to Figs. and . a – c , Phosphorylation Site Preference (PSP) plots showing single-cell correlation of Rb phosphorylation at different sites in RPE-1 cells ( a ), BJ-5ta cells ( b ), or U2OS cells ( c ). Each phosphorylation signal was normalized by the total Rb antibody signal in the same cell and each axis was adjusted to the average phosphorylation signal in S phase of 1. Color indicates relative cell population density. A red line shows fitting with a preferential relative phosphorylation or dephosphorylation rate between the two sites (PSP coeff ) (see more details in Methods). Rb phosphorylation at T373 or T826 was plotted against S807/S811. a , b , Cells were fixed 8 h after release with growth media ( a : n = 5019, 4900 cells for T373, T826, respectively. b : n = 3409, 3502 cells for T373, T826, respectively. 1 of n = 3 biological replicates). c , Cells asynchronously cycling in growth media were fixed 6 h after DMSO or CDK4/6i (100 nM) treatment (DMSO: n = 704, 700 cells for T373, T826, respectively. CDK4/6i: n = 850, 878 cells for T373, T826, respectively. 1 of n = 3 biological replicates). d , Single-cell correlation of CDK2 activity and Rb phosphorylation at T373 and T826. Cells were fixed 48 h after release with starvation media + EGF (20 ng/mL) + CDK4/6i (1 μM). Rb phosphorylation levels as a function CDK2 activity were fitted by sigmoidal curves (sigmoidal curves with Hill coefficients. n = 1967, 2050 cells for T373, T826, respectively).
    Figure Legend Snippet: Related to Figs. and . a – c , Phosphorylation Site Preference (PSP) plots showing single-cell correlation of Rb phosphorylation at different sites in RPE-1 cells ( a ), BJ-5ta cells ( b ), or U2OS cells ( c ). Each phosphorylation signal was normalized by the total Rb antibody signal in the same cell and each axis was adjusted to the average phosphorylation signal in S phase of 1. Color indicates relative cell population density. A red line shows fitting with a preferential relative phosphorylation or dephosphorylation rate between the two sites (PSP coeff ) (see more details in Methods). Rb phosphorylation at T373 or T826 was plotted against S807/S811. a , b , Cells were fixed 8 h after release with growth media ( a : n = 5019, 4900 cells for T373, T826, respectively. b : n = 3409, 3502 cells for T373, T826, respectively. 1 of n = 3 biological replicates). c , Cells asynchronously cycling in growth media were fixed 6 h after DMSO or CDK4/6i (100 nM) treatment (DMSO: n = 704, 700 cells for T373, T826, respectively. CDK4/6i: n = 850, 878 cells for T373, T826, respectively. 1 of n = 3 biological replicates). d , Single-cell correlation of CDK2 activity and Rb phosphorylation at T373 and T826. Cells were fixed 48 h after release with starvation media + EGF (20 ng/mL) + CDK4/6i (1 μM). Rb phosphorylation levels as a function CDK2 activity were fitted by sigmoidal curves (sigmoidal curves with Hill coefficients. n = 1967, 2050 cells for T373, T826, respectively).

    Techniques Used: De-Phosphorylation Assay, Activity Assay

    ant pr 1 hygromycin b gold invivogen  (InvivoGen)


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    ATCC ml 1 hygromycin b u2os human osteosarcoma epithelial cells
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    Related to Figs. and . a – c , Phosphorylation Site Preference (PSP) plots showing single-cell correlation of Rb phosphorylation at different sites in RPE-1 cells ( a ), BJ-5ta cells ( b ), or <t>U2OS</t> cells ( c ). Each phosphorylation signal was normalized by the total Rb antibody signal in the same cell and each axis was adjusted to the average phosphorylation signal in S phase of 1. Color indicates relative cell population density. A red line shows fitting with a preferential relative phosphorylation or dephosphorylation rate between the two sites (PSP coeff ) (see more details in Methods). Rb phosphorylation at T373 or T826 was plotted against S807/S811. a , b , Cells were fixed 8 h after release with growth media ( a : n = 5019, 4900 cells for T373, T826, respectively. b : n = 3409, 3502 cells for T373, T826, respectively. 1 of n = 3 biological replicates). c , Cells asynchronously cycling in growth media were fixed 6 h after DMSO or CDK4/6i (100 nM) treatment (DMSO: n = 704, 700 cells for T373, T826, respectively. CDK4/6i: n = 850, 878 cells for T373, T826, respectively. 1 of n = 3 biological replicates). d , Single-cell correlation of CDK2 activity and Rb phosphorylation at T373 and T826. Cells were fixed 48 h after release with starvation media + EGF (20 ng/mL) + CDK4/6i (1 μM). Rb phosphorylation levels as a function CDK2 activity were fitted by sigmoidal curves (sigmoidal curves with Hill coefficients. n = 1967, 2050 cells for T373, T826, respectively).
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    Related to Figs. and . a – c , Phosphorylation Site Preference (PSP) plots showing single-cell correlation of Rb phosphorylation at different sites in RPE-1 cells ( a ), BJ-5ta cells ( b ), or <t>U2OS</t> cells ( c ). Each phosphorylation signal was normalized by the total Rb antibody signal in the same cell and each axis was adjusted to the average phosphorylation signal in S phase of 1. Color indicates relative cell population density. A red line shows fitting with a preferential relative phosphorylation or dephosphorylation rate between the two sites (PSP coeff ) (see more details in Methods). Rb phosphorylation at T373 or T826 was plotted against S807/S811. a , b , Cells were fixed 8 h after release with growth media ( a : n = 5019, 4900 cells for T373, T826, respectively. b : n = 3409, 3502 cells for T373, T826, respectively. 1 of n = 3 biological replicates). c , Cells asynchronously cycling in growth media were fixed 6 h after DMSO or CDK4/6i (100 nM) treatment (DMSO: n = 704, 700 cells for T373, T826, respectively. CDK4/6i: n = 850, 878 cells for T373, T826, respectively. 1 of n = 3 biological replicates). d , Single-cell correlation of CDK2 activity and Rb phosphorylation at T373 and T826. Cells were fixed 48 h after release with starvation media + EGF (20 ng/mL) + CDK4/6i (1 μM). Rb phosphorylation levels as a function CDK2 activity were fitted by sigmoidal curves (sigmoidal curves with Hill coefficients. n = 1967, 2050 cells for T373, T826, respectively).
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    Related to Figs. and . a – c , Phosphorylation Site Preference (PSP) plots showing single-cell correlation of Rb phosphorylation at different sites in RPE-1 cells ( a ), BJ-5ta cells ( b ), or <t>U2OS</t> cells ( c ). Each phosphorylation signal was normalized by the total Rb antibody signal in the same cell and each axis was adjusted to the average phosphorylation signal in S phase of 1. Color indicates relative cell population density. A red line shows fitting with a preferential relative phosphorylation or dephosphorylation rate between the two sites (PSP coeff ) (see more details in Methods). Rb phosphorylation at T373 or T826 was plotted against S807/S811. a , b , Cells were fixed 8 h after release with growth media ( a : n = 5019, 4900 cells for T373, T826, respectively. b : n = 3409, 3502 cells for T373, T826, respectively. 1 of n = 3 biological replicates). c , Cells asynchronously cycling in growth media were fixed 6 h after DMSO or CDK4/6i (100 nM) treatment (DMSO: n = 704, 700 cells for T373, T826, respectively. CDK4/6i: n = 850, 878 cells for T373, T826, respectively. 1 of n = 3 biological replicates). d , Single-cell correlation of CDK2 activity and Rb phosphorylation at T373 and T826. Cells were fixed 48 h after release with starvation media + EGF (20 ng/mL) + CDK4/6i (1 μM). Rb phosphorylation levels as a function CDK2 activity were fitted by sigmoidal curves (sigmoidal curves with Hill coefficients. n = 1967, 2050 cells for T373, T826, respectively).
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    Related to Figs. and . a – c , Phosphorylation Site Preference (PSP) plots showing single-cell correlation of Rb phosphorylation at different sites in RPE-1 cells ( a ), BJ-5ta cells ( b ), or <t>U2OS</t> cells ( c ). Each phosphorylation signal was normalized by the total Rb antibody signal in the same cell and each axis was adjusted to the average phosphorylation signal in S phase of 1. Color indicates relative cell population density. A red line shows fitting with a preferential relative phosphorylation or dephosphorylation rate between the two sites (PSP coeff ) (see more details in Methods). Rb phosphorylation at T373 or T826 was plotted against S807/S811. a , b , Cells were fixed 8 h after release with growth media ( a : n = 5019, 4900 cells for T373, T826, respectively. b : n = 3409, 3502 cells for T373, T826, respectively. 1 of n = 3 biological replicates). c , Cells asynchronously cycling in growth media were fixed 6 h after DMSO or CDK4/6i (100 nM) treatment (DMSO: n = 704, 700 cells for T373, T826, respectively. CDK4/6i: n = 850, 878 cells for T373, T826, respectively. 1 of n = 3 biological replicates). d , Single-cell correlation of CDK2 activity and Rb phosphorylation at T373 and T826. Cells were fixed 48 h after release with starvation media + EGF (20 ng/mL) + CDK4/6i (1 μM). Rb phosphorylation levels as a function CDK2 activity were fitted by sigmoidal curves (sigmoidal curves with Hill coefficients. n = 1967, 2050 cells for T373, T826, respectively).
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    Related to Figs. and . a – c , Phosphorylation Site Preference (PSP) plots showing single-cell correlation of Rb phosphorylation at different sites in RPE-1 cells ( a ), BJ-5ta cells ( b ), or <t>U2OS</t> cells ( c ). Each phosphorylation signal was normalized by the total Rb antibody signal in the same cell and each axis was adjusted to the average phosphorylation signal in S phase of 1. Color indicates relative cell population density. A red line shows fitting with a preferential relative phosphorylation or dephosphorylation rate between the two sites (PSP coeff ) (see more details in Methods). Rb phosphorylation at T373 or T826 was plotted against S807/S811. a , b , Cells were fixed 8 h after release with growth media ( a : n = 5019, 4900 cells for T373, T826, respectively. b : n = 3409, 3502 cells for T373, T826, respectively. 1 of n = 3 biological replicates). c , Cells asynchronously cycling in growth media were fixed 6 h after DMSO or CDK4/6i (100 nM) treatment (DMSO: n = 704, 700 cells for T373, T826, respectively. CDK4/6i: n = 850, 878 cells for T373, T826, respectively. 1 of n = 3 biological replicates). d , Single-cell correlation of CDK2 activity and Rb phosphorylation at T373 and T826. Cells were fixed 48 h after release with starvation media + EGF (20 ng/mL) + CDK4/6i (1 μM). Rb phosphorylation levels as a function CDK2 activity were fitted by sigmoidal curves (sigmoidal curves with Hill coefficients. n = 1967, 2050 cells for T373, T826, respectively).
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    Related to Figs. and . a – c , Phosphorylation Site Preference (PSP) plots showing single-cell correlation of Rb phosphorylation at different sites in RPE-1 cells ( a ), BJ-5ta cells ( b ), or <t>U2OS</t> cells ( c ). Each phosphorylation signal was normalized by the total Rb antibody signal in the same cell and each axis was adjusted to the average phosphorylation signal in S phase of 1. Color indicates relative cell population density. A red line shows fitting with a preferential relative phosphorylation or dephosphorylation rate between the two sites (PSP coeff ) (see more details in Methods). Rb phosphorylation at T373 or T826 was plotted against S807/S811. a , b , Cells were fixed 8 h after release with growth media ( a : n = 5019, 4900 cells for T373, T826, respectively. b : n = 3409, 3502 cells for T373, T826, respectively. 1 of n = 3 biological replicates). c , Cells asynchronously cycling in growth media were fixed 6 h after DMSO or CDK4/6i (100 nM) treatment (DMSO: n = 704, 700 cells for T373, T826, respectively. CDK4/6i: n = 850, 878 cells for T373, T826, respectively. 1 of n = 3 biological replicates). d , Single-cell correlation of CDK2 activity and Rb phosphorylation at T373 and T826. Cells were fixed 48 h after release with starvation media + EGF (20 ng/mL) + CDK4/6i (1 μM). Rb phosphorylation levels as a function CDK2 activity were fitted by sigmoidal curves (sigmoidal curves with Hill coefficients. n = 1967, 2050 cells for T373, T826, respectively).
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    Related to Figs. and . a – c , Phosphorylation Site Preference (PSP) plots showing single-cell correlation of Rb phosphorylation at different sites in RPE-1 cells ( a ), BJ-5ta cells ( b ), or U2OS cells ( c ). Each phosphorylation signal was normalized by the total Rb antibody signal in the same cell and each axis was adjusted to the average phosphorylation signal in S phase of 1. Color indicates relative cell population density. A red line shows fitting with a preferential relative phosphorylation or dephosphorylation rate between the two sites (PSP coeff ) (see more details in Methods). Rb phosphorylation at T373 or T826 was plotted against S807/S811. a , b , Cells were fixed 8 h after release with growth media ( a : n = 5019, 4900 cells for T373, T826, respectively. b : n = 3409, 3502 cells for T373, T826, respectively. 1 of n = 3 biological replicates). c , Cells asynchronously cycling in growth media were fixed 6 h after DMSO or CDK4/6i (100 nM) treatment (DMSO: n = 704, 700 cells for T373, T826, respectively. CDK4/6i: n = 850, 878 cells for T373, T826, respectively. 1 of n = 3 biological replicates). d , Single-cell correlation of CDK2 activity and Rb phosphorylation at T373 and T826. Cells were fixed 48 h after release with starvation media + EGF (20 ng/mL) + CDK4/6i (1 μM). Rb phosphorylation levels as a function CDK2 activity were fitted by sigmoidal curves (sigmoidal curves with Hill coefficients. n = 1967, 2050 cells for T373, T826, respectively).

    Journal: Nature

    Article Title: An intermediate Rb–E2F activity state safeguards proliferation commitment

    doi: 10.1038/s41586-024-07554-2

    Figure Lengend Snippet: Related to Figs. and . a – c , Phosphorylation Site Preference (PSP) plots showing single-cell correlation of Rb phosphorylation at different sites in RPE-1 cells ( a ), BJ-5ta cells ( b ), or U2OS cells ( c ). Each phosphorylation signal was normalized by the total Rb antibody signal in the same cell and each axis was adjusted to the average phosphorylation signal in S phase of 1. Color indicates relative cell population density. A red line shows fitting with a preferential relative phosphorylation or dephosphorylation rate between the two sites (PSP coeff ) (see more details in Methods). Rb phosphorylation at T373 or T826 was plotted against S807/S811. a , b , Cells were fixed 8 h after release with growth media ( a : n = 5019, 4900 cells for T373, T826, respectively. b : n = 3409, 3502 cells for T373, T826, respectively. 1 of n = 3 biological replicates). c , Cells asynchronously cycling in growth media were fixed 6 h after DMSO or CDK4/6i (100 nM) treatment (DMSO: n = 704, 700 cells for T373, T826, respectively. CDK4/6i: n = 850, 878 cells for T373, T826, respectively. 1 of n = 3 biological replicates). d , Single-cell correlation of CDK2 activity and Rb phosphorylation at T373 and T826. Cells were fixed 48 h after release with starvation media + EGF (20 ng/mL) + CDK4/6i (1 μM). Rb phosphorylation levels as a function CDK2 activity were fitted by sigmoidal curves (sigmoidal curves with Hill coefficients. n = 1967, 2050 cells for T373, T826, respectively).

    Article Snippet: BJ-5ta human foreskin fibroblast cells (ATCC, CRL-4001, RRID: CVCL_6573 ) were cultured in DMEM growth medium (Gibco, 11995065) supplemented with 20% Medium 199 (Thermo Fisher, 11150059), 10% FBS and 0.01 mg ml –1 hygromycin B. U2OS human osteosarcoma epithelial cells (ATCC, HTB-96, RRID: CVCL_0042 ) and Lenti-X 293T human embryonic kidney cells (Takara Bio, 632180, RRID: CVCL_4401 ) were cultured in DMEM growth medium with 10% FBS.

    Techniques: De-Phosphorylation Assay, Activity Assay