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sch900776 mk8776  (MedChemExpress)
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MedChemExpress sch900776 mk8776
a Schematic diagram of the setup of the co-culture competition assay. b UT-7/Tpo CALR wild type and del61/WT cell line were incubated with 0.8 µM of CHK1 inhibitor rabusertib or 1 µM of <t>MK8776</t> over 12 days. Wild type (mCherry negative) and mutant cells (mCherry positive) were mixed at a 1:1 ratio and 1% of TPO conditioned media was present. Samples were collected every three days and the ratio of wild-type cells to mutant cells was measured by FACs analysis. c Ba/F3-MPL CALR wild type and del37/del37 cell line were incubated with 0.8 µM rabusertib or 1 µM MK8776 over 5 days. Wild type (mCherry negative) and mutant cells (mCherry positive) were mixed at a 1:1 ratio and 1% of TPO conditioned media was present. Samples were collected at day 0, day 2, and the end of treatment. The ratio of wild-type cells to mutant cells was measured by FACs analysis. d Ba/F3-MPL CALR wild type and CALR knockout cell line were incubated with 1 µM MK8776 over 5 days in a co-culture competition assay. e Ba/F3-MPL JAK2 wild type and JAK2V617F mutated cell lines were incubated with 1 µM MK8776 over 5 days in a co-culture competition assay.
Sch900776 Mk8776, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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a Schematic diagram of the setup of the co-culture competition assay. b UT-7/Tpo CALR wild type and del61/WT cell line were incubated with 0.8 µM of CHK1 inhibitor rabusertib or 1 µM of MK8776 over 12 days. Wild type (mCherry negative) and mutant cells (mCherry positive) were mixed at a 1:1 ratio and 1% of TPO conditioned media was present. Samples were collected every three days and the ratio of wild-type cells to mutant cells was measured by FACs analysis. c Ba/F3-MPL CALR wild type and del37/del37 cell line were incubated with 0.8 µM rabusertib or 1 µM MK8776 over 5 days. Wild type (mCherry negative) and mutant cells (mCherry positive) were mixed at a 1:1 ratio and 1% of TPO conditioned media was present. Samples were collected at day 0, day 2, and the end of treatment. The ratio of wild-type cells to mutant cells was measured by FACs analysis. d Ba/F3-MPL CALR wild type and CALR knockout cell line were incubated with 1 µM MK8776 over 5 days in a co-culture competition assay. e Ba/F3-MPL JAK2 wild type and JAK2V617F mutated cell lines were incubated with 1 µM MK8776 over 5 days in a co-culture competition assay.

Journal: Blood Cancer Journal

Article Title: High-throughput drug screening identifies the ATR-CHK1 pathway as a therapeutic vulnerability of CALR mutated hematopoietic cells

doi: 10.1038/s41408-021-00531-2

Figure Lengend Snippet: a Schematic diagram of the setup of the co-culture competition assay. b UT-7/Tpo CALR wild type and del61/WT cell line were incubated with 0.8 µM of CHK1 inhibitor rabusertib or 1 µM of MK8776 over 12 days. Wild type (mCherry negative) and mutant cells (mCherry positive) were mixed at a 1:1 ratio and 1% of TPO conditioned media was present. Samples were collected every three days and the ratio of wild-type cells to mutant cells was measured by FACs analysis. c Ba/F3-MPL CALR wild type and del37/del37 cell line were incubated with 0.8 µM rabusertib or 1 µM MK8776 over 5 days. Wild type (mCherry negative) and mutant cells (mCherry positive) were mixed at a 1:1 ratio and 1% of TPO conditioned media was present. Samples were collected at day 0, day 2, and the end of treatment. The ratio of wild-type cells to mutant cells was measured by FACs analysis. d Ba/F3-MPL CALR wild type and CALR knockout cell line were incubated with 1 µM MK8776 over 5 days in a co-culture competition assay. e Ba/F3-MPL JAK2 wild type and JAK2V617F mutated cell lines were incubated with 1 µM MK8776 over 5 days in a co-culture competition assay.

Article Snippet: For other assays, the following drugs and suppliers were used: Rabusertib/LY2603618 (Cayman Chemical Company, USA); SCH900776/MK8776 (MedChemExpress, USA); Adavosertib/MK-1775 (MedChemExpress, USA).

Techniques: Co-Culture Assay, Competitive Binding Assay, Incubation, Mutagenesis, Knock-Out

a Western blot analysis of UT-7/Tpo CALR WT and CALR del61/WT whole cell lysates following treatment of the indicated drugs for 24 h. 1 µM MK8776, 0.8 µM rabusertib, and 0.3 µM adavosertib were used as single-agent treatment. 0.2 µM adavosertib combined with 0.17 µM MK8776 or 0.13 µM rabusertib were used for combination treatment. b Top panel: Immunofluorescence of UT-7/Tpo CALR WT and CALR del61/WT following 1 µM MK8776 treatment for 6 h. Pseudocolor annotation: red represents TUNEL; green represents γ-H2AX; blue represents DAPI. Bottom panel: Quantification of γ-H2AX positive cells in UT-7/Tpo CALR WT and CALR del61/WT cells after 6 and 24 h treatment of MK8776. * p < 0.05; **** p < 0.0001.

Journal: Blood Cancer Journal

Article Title: High-throughput drug screening identifies the ATR-CHK1 pathway as a therapeutic vulnerability of CALR mutated hematopoietic cells

doi: 10.1038/s41408-021-00531-2

Figure Lengend Snippet: a Western blot analysis of UT-7/Tpo CALR WT and CALR del61/WT whole cell lysates following treatment of the indicated drugs for 24 h. 1 µM MK8776, 0.8 µM rabusertib, and 0.3 µM adavosertib were used as single-agent treatment. 0.2 µM adavosertib combined with 0.17 µM MK8776 or 0.13 µM rabusertib were used for combination treatment. b Top panel: Immunofluorescence of UT-7/Tpo CALR WT and CALR del61/WT following 1 µM MK8776 treatment for 6 h. Pseudocolor annotation: red represents TUNEL; green represents γ-H2AX; blue represents DAPI. Bottom panel: Quantification of γ-H2AX positive cells in UT-7/Tpo CALR WT and CALR del61/WT cells after 6 and 24 h treatment of MK8776. * p < 0.05; **** p < 0.0001.

Article Snippet: For other assays, the following drugs and suppliers were used: Rabusertib/LY2603618 (Cayman Chemical Company, USA); SCH900776/MK8776 (MedChemExpress, USA); Adavosertib/MK-1775 (MedChemExpress, USA).

Techniques: Western Blot, Immunofluorescence, TUNEL Assay

a Left panel: histogram of UT-7/Tpo CALR WT and CALR del61/WT cell lines treated with 1 µM MK8776 or DMSO for 24 h, followed by DAPI staining to indicate DNA content. The percentage of cells at pre-G1, G1, S, G2-M phases is labeled in the graphs. Right panel: FACS co-staining of DAPI and γ-H2AX at a 24 h time point post-treatment. γ-H2AX positive S-phase cells are labeled and the percentage of this population is indicated in the graphs. b Left panel: histogram of UT-7/Tpo CALR WT and CALR del61/WT cell lines treated with 1 µM MK8776 or DMSO for 72 h, followed by DAPI staining to indicate DNA content. The percentage of cells at pre-G1, G1, S, G2-M phases is labeled in the graphs. Right panel: FACS co-staining of DAPI and γ-H2AX at a 72 h time point post-treatment. γ-H2AX positive S-phase cells are labeled and the percentage of this population is indicated in the graphs.

Journal: Blood Cancer Journal

Article Title: High-throughput drug screening identifies the ATR-CHK1 pathway as a therapeutic vulnerability of CALR mutated hematopoietic cells

doi: 10.1038/s41408-021-00531-2

Figure Lengend Snippet: a Left panel: histogram of UT-7/Tpo CALR WT and CALR del61/WT cell lines treated with 1 µM MK8776 or DMSO for 24 h, followed by DAPI staining to indicate DNA content. The percentage of cells at pre-G1, G1, S, G2-M phases is labeled in the graphs. Right panel: FACS co-staining of DAPI and γ-H2AX at a 24 h time point post-treatment. γ-H2AX positive S-phase cells are labeled and the percentage of this population is indicated in the graphs. b Left panel: histogram of UT-7/Tpo CALR WT and CALR del61/WT cell lines treated with 1 µM MK8776 or DMSO for 72 h, followed by DAPI staining to indicate DNA content. The percentage of cells at pre-G1, G1, S, G2-M phases is labeled in the graphs. Right panel: FACS co-staining of DAPI and γ-H2AX at a 72 h time point post-treatment. γ-H2AX positive S-phase cells are labeled and the percentage of this population is indicated in the graphs.

Article Snippet: For other assays, the following drugs and suppliers were used: Rabusertib/LY2603618 (Cayman Chemical Company, USA); SCH900776/MK8776 (MedChemExpress, USA); Adavosertib/MK-1775 (MedChemExpress, USA).

Techniques: Staining, Labeling