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Hs00153408 m1

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Hs00153408_m1 is a TaqMan Gene Expression Assay provided by Thermo Fisher Scientific. It is designed to detect and quantify the expression of a specific gene target. The assay includes a pre-designed and pre-optimized set of primers and a probe to enable accurate and reliable gene expression analysis.

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Lab products found in correlation

Oligo dt primer, by Promega (3 mentions) Hs00984230 m1, by Thermo Fisher Scientific (3 mentions) M mlv reverse transcriptase, by Promega (3 mentions) Hoxa9, by Thermo Fisher Scientific (2 mentions) Hs00365956 m1, by Thermo Fisher Scientific (2 mentions) Meis1, by Thermo Fisher Scientific (2 mentions) Hs00180020 m1, by Thermo Fisher Scientific (2 mentions) Rna extraction kit, by Promega (1 mentions) Quantstudio 7 flex real time pcr system, by Thermo Fisher Scientific (1 mentions) Purelink rna mini kit, by Thermo Fisher Scientific (1 mentions) Taqman gene expression master mix, by Thermo Fisher Scientific (1 mentions) Rna extraction kit, by Qiagen (1 mentions)

6 protocols using hs00153408 m1

1

In Vivo Evaluation of Compound Efficacy Against MV4;11 Tumor Xenografts

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Female Balb/c nude mice (6–8 weeks, Beijing Anikeeper Biotech. Ltd) were inoculated subcutaneously on the right flank with MV4;11 tumor cells (1 × 107) in 0.1 mL of 1:1 v/v of IMDM:Matrigel for tumor development. Compound treatment was initiated when the mean tumor volume reached about 300 to 400 mm3. Mice were randomly assigned to respective treatment groups such that the average starting tumor size and body weight were the same for each treatment group. Mice were dosed with vehicle (5% DMSO:95% of 10% w/v Kleptose HPB, pH 4.0) or compound at 100 or 200 mg/kg p.o., b.i.d. at 12 hours intervals. For PD readouts (tumor volume and gene analysis), five animals per group were used. Animals were checked daily and body weights measured every 2 days throughout the study. Tumor volume was estimated using the formula: TV = a × b2/2 throughout the study, where “a” and “b” are the long and short diameters of a tumor, respectively. Individual tumor samples for gene analysis were taken at 2 hours after dose 8 days of dosing, and the following genes were analyzed: MYC (Hs00153408_m1, Thermo Fisher), HOXA9 (Hs00365956_m1, Thermo Fisher), MEIS1 (Hs00180020_m1, Thermo Fisher), and MYB (see Supplementary Table S4).
Liu Y., Li Q., Alikarami F., Barrett D.R., Mahdavi L., Li H., Tang S., Khan T.A., Michino M., Hill C., Song L., Yang L., Li Y., Pokharel S.P., Stamford A.W., Liverton N., Renzetti L.M., Taylor S., Watt G.F., Ladduwahetty T., Kargman S., Meinke P.T., Foley M.A., Shi J., Li H., Carroll M., Chen C.W., Gardini A., Maillard I., Huggins D.J., Bernt K.M, & Wan L. (2022). Small-Molecule Inhibition of the Acyl-Lysine Reader ENL as a Strategy against Acute Myeloid Leukemia. Cancer Discovery, 12(11), 2684-2709.
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2

Balb/c Nude Mice Xenograft Model

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Female Balb/c nude mice (6–8 weeks, Beijing Anikeeper Biotech. Ltd) were inoculated subcutaneously on the right flank with MV4–11 tumor cells (1 × 107) in 0.1 mL of 1:1 v/v of IMDM: Matrigel for tumor development. Compound treatment was initiated when the mean tumor volume reaches about 300~400 mm3. Mice were randomly assigned to respective treatment groups such that the average starting tumor size and body weight was the same for each treatment group. Mice were dosed with vehicle (5% DMSO & 95% (10% HPB in water) v/v adjusted to pH4.0) or compound at 100 or 200mg/kg P.O., BID at 12 hours intervals. For PD readouts (tumor volume ad gene analysis), 5 animals per group were used. Animals were checked daily and body weights measure every 2 days throughout the study. Tumor volume was estimated using the formula: TV = a × b2/2 throughout the study, where “a” and “b” are the long and short diameters of a tumor, respectively. Individual tumor samples for gene analysis were taken at 2 hours post-dose 8 days of dosing and the following genes were analyzed: MYC (Hs00153408_m1, ThermoFisher), HOXA9 (Hs00365956_m1, Thermofisher), MEIS1 (Hs00180020_m1, Thermofisher), MYB (see Supplementary Table S4).
Liu Y., Li Q., Alikarami F., Barrett D.R., Mahdavi L., Li H., Tang S., Khan T.A., Michino M., Hill C., Song L., Yang L., Li Y., Pokharel S.P., Stamford A.W., Liverton N., Renzetti L.M., Taylor S., Watt G.F., Ladduwahetty T., Kargman S., Meinke P.T., Foley M.A., Shi J., Li H., Carroll M., Chen C.W., Gardini A., Maillard I., Huggins D.J., Bernt K.M, & Wan L. (2022). Small-Molecule Inhibition of the Acyl-Lysine Reader ENL as a Strategy against Acute Myeloid Leukemia. Cancer Discovery, 12(11), 2684-2709.
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3

Quantifying mRNA Expression by qPCR

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Total mRNA were isolated by phenol/chloroform isolation. cDNA synthesis was performed using MMLV reverse transcriptase and oligo-dT primers (both Promega, Southampton, UK). MYC, PDCD4 and B2M mRNA expression was quantified by Q-PCR using probes Hs00153408_m1, Hs00377253_m1 and Hs00984230_m1, respectively (Life Technologies, Carlsbad, US). mRNA abundance was determined for each mRNA against a standard curve, providing cDNA values and relative mRNA expression was calculated by normalizing the obtained values against B2M mRNA.
Taylor J., Wilmore S., Marriot S., Rogers-Broadway K.R., Fell R., Minton A.R., Branch T., Ashton-Key M., Coldwell M., Stevenson F.K., Forconi F., Steele A.J., Packham G, & Yeomans A. (2022). B-cell receptor signaling induces proteasomal degradation of PDCD4 via MEK1/2 and mTORC1 in malignant B cells. Cellular Signalling, 94, 110311.
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4

Quantifying MYC, MCL1 mRNA Levels

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Total mRNA or fractions from polysome profiling were isolated using the Promega RNA extraction kit (Promega, Southampton, UK). cDNA synthesis was performed using MMLV reverse transcriptase and oligo-dT primers (both Promega, Southampton, UK). MYC, MCL1 and B2M mRNA expression was quantified by Q-PCR using probes Hs00153408_m1, Hs00172036_m1 and Hs00984230_m1, respectively (Life Technologies). mRNA abundance was determined for each mRNA against a standard curve, providing cDNA values and relative mRNA expression was calculated by normalizing the obtained values against B2M mRNA.
Wilmore S., Rogers-Broadway K.R., Taylor J., Lemm E., Fell R., Stevenson F.K., Forconi F., Steele A.J., Coldwell M., Packham G, & Yeomans A. (2021). Targeted inhibition of eIF4A suppresses B-cell receptor-induced translation and expression of MYC and MCL1 in chronic lymphocytic leukemia cells. Cellular and Molecular Life Sciences, 78(17-18), 6337-6349.
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5

Quantitative Analysis of Immune Genes

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RNA was extracted using Purelink RNA mini kit (Life Technologies) per standard techniques. Real-time PCR was performed using TaqMan Gene Expression Master Mix (Life Technologies) in a QuantStudio 7 Flex Real-Time PCR System (Thermo Fisher Scientific), according to the manufacturer’s instructions. Each of the reactions was performed in triplicate, including no template controls and amplification of a housekeeping gene, GAPDH. Gene-specific assays were Mm00452054_m1 for mouse PD-L1, Mm00803857_m1 for mouse n-myc, Mm00487804_m1 for mouse c-myc, Mm00775963_g1 for mouse Id1, Mm00711781_m1 for mouse Id2, Mm00492575_m1 for mouse Id3, Mm00499701_m1 for mouse Id4, Mm99999915_g1 for mouse Gapdh, Hs00232074_m1 for human MYCN, Hs00153408_m1 for human c-MYC, Hs00204257_m1 for human PD-L1 and Hs02786624_g1 for human GAPDH (Life Technologies). Changes in relative gene expression normalized to GAPDH levels were determined using the ΔΔCt method. Results were averaged and statistically analyzed using t-tests.
Wu X., Nelson M., Basu M., Srinivasan P., Lazarski C., Zhang P., Zheng P, & Sandler A.D. (2021). MYC oncogene is associated with suppression of tumor immunity and targeting Myc induces tumor cell immunogenicity for therapeutic whole cell vaccination. Journal for Immunotherapy of Cancer, 9(3), e001388.
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6

Quantitative mRNA Expression Analysis

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Total mRNA was isolated using the Qiagen RNA extraction kit and cDNA synthesis was performed using MMLV reverse transcriptase and oligo-dT primers (both Promega, Southampton, UK). MYC, CCND2, BCL2A1 and B2M mRNA expression was quantified by Q-PCR using probes Hs00153408_m1, Hs00277041_m1, Hs00187845_m1 and Hs00984230_m1, respectively (Life Technologies). RNA abundance was determined for each RNA against a standard curve, providing cDNA values and relative RNA expression was calculated by normalizing the obtained values against B2M expression.
Arthur R., Wathen A., Lemm E.A., Stevenson F.K., Forconi F., Linley A.J., Steele A.J., Packham G, & Valle-Argos B. (2022). BTK-independent regulation of calcium signalling downstream of the B-cell receptor in malignant B-cells. Cellular signalling, 96.
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