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Pd0325901

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PD0325901 is a selective and potent MEK inhibitor. It inhibits the activation of MEK1 and MEK2, which are key components of the MAPK/ERK signaling pathway.

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Lab products found in correlation

Chir99021, by LC Laboratories (11 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) Rpmi 1640, by Thermo Fisher Scientific (3 mentions) Lapatinib, by LC Laboratories (2 mentions) Gdc 0941, by LC Laboratories (2 mentions) Mk2206, by LC Laboratories (2 mentions) Mln8237, by LC Laboratories (2 mentions) Lee011, by LC Laboratories (2 mentions) Mda mb 231, by American Type Culture Collection (2 mentions) Mcf 7, by American Type Culture Collection (2 mentions) Hcc38, by American Type Culture Collection (2 mentions) Bt474, by American Type Culture Collection (2 mentions) Mda mb 453, by American Type Culture Collection (2 mentions) Bt 20, by American Type Culture Collection (2 mentions) Hcc1428, by American Type Culture Collection (2 mentions) Sum52pe, by American Type Culture Collection (2 mentions) Horse serum, by Thermo Fisher Scientific (2 mentions) Recombinant bmp4, by Thermo Fisher Scientific (2 mentions) Hek293t cell, by American Type Culture Collection (2 mentions) Fibroblast growth factor 2 (fgf2), by Thermo Fisher Scientific (2 mentions)

25 protocols using pd0325901

1

Expansion of Feeder-free Mouse ESCs

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ES cells were cultured in ESC media (15% FBS, 25 mM HEPES, 1× GlutaMAX, 1× MEM Non-essential Amino Acids Solution, 1× Penicillin/Streptomycin, and 0.055 mM β-Mercaptoethanol in DMEM High Glucose (4.5 g/L)) containing 2i (1 μM PD0325901, LC Laboratories; and 3μM CHIR99021, LC Laboratories) and LIF (1300 U/mL, in-house) on cell culture plates coated with 0.2% gelatin under feeder-free conditions. The expanded ES colonies were dissociated using 0.25% trypsin-EDTA solution for passaging.
Maezawa S., Sakashita A., Yukawa M., Chen X., Takahashi K., Alavattam K.G., Nakata I., Weirauch M.T., Barski A, & Namekawa S.H. (2020). Super-enhancer switching drives a burst in gene expression at the mitosis-to-meiosis transition. Nature structural & molecular biology, 27(10), 978-988.
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2

Culture of Mouse Embryonic Stem Cells

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The mouse ESC line AB2.2 was cultured on mouse embryonic fibroblast (MEF) feeder cells with ES cell medium supplemented with 15% fetal bovine serum (Hyclone, Logan, UT, USA, cat. no. SH30396.03), 1000 U/mL leukemia inhibitory factor (LIF, Millipore, Burlington, MA, USA, cat. no. ESG1107), 3 μM CHIR99021 (LC Laboratories, Woburn, MA, USA, cat. no. C-6556), and 1 μM PD0325901 (LC Laboratories, cat. no. P-9688). Fifty millimolar lactate (Sangon Biotech, Shanghai, China, cat. no. A604046) was added and maintained for 24 h before experimental procedures. Cells were cultured in a humidified chamber at 37 °C with 5% CO2. For culture of mouse ESC lines, the medium was refreshed daily, and cells were routinely passaged every 2 days.
Tian Q, & Zhou L.Q. (2022). Lactate Activates Germline and Cleavage Embryo Genes in Mouse Embryonic Stem Cells. Cells, 11(3), 548.
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3

Breast Cancer Cell Line Characterization

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BT549 and SKBR3 cells were obtained from the UCSF Cell Culture Facility.
BT20, BT474, HCC1428, HCC38, LY2, MCF7, MDAMB231, MDAMB453, T47D, SUM52PE, and
ZR75B cell lines were obtained from the American Type Culture Collection (ATCC).
Cell lines used for proteomic profiling and molecular analyses were
authenticated by STR analysis. Lines were grown according to published
protocols50 except for
SKBR3 which was cultured using RPMI media supplemented with 10% fetal
bovine serum (FBS) and 1% pen/strep. All cell lines tested negative for
mycoplasma contamination. Drugs used for cell culture experiments in this study
were purchased from Selleck Chemicals (GDC-0941, MK2206, PD0325901, Lapatinib,
MLN8237, and LEE011) and LC Laboratories (RAD001).
Donnella H.J., Webber J.T., Levin R.S., Camarda R., Momcilovic O., Bayani N., Shah K.N., Korkola J., Shokat K.M., Goga A., Gordan J.D, & Bandyopadhyay S. (2018). Kinome rewiring reveals AURKA limits PI3K-pathway inhibitor efficacy in breast cancer. Nature chemical biology, 14(8), 768-777.
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4

Generating Xist Deletion Mutant ES Cells

Cited 1 time
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16.7 female ES cells were used to generate Xist repeat E deletion mutant [30 (link)]. ES cells were grown under standard conditions as described except for the addition of 2i inhibitors (3 μM CHIR99021 and 1 μM PD0325901, LC laboratories) [28 (link)]. Xist expression from tet-inducible Xist cDNA transgene in T20 cell lines was induced by adding 1 μg/ml doxycycline (Dox) in the culture medium for differentiation for 2 days [21 (link)]. Cre-LoxP mediated insertion of the inducible Xist cDNA transgenes was done as described [21 (link)].
Yue M., Ogawa A., Yamada N., Charles Richard J.L., Barski A, & Ogawa Y. (2017). Xist RNA repeat E is essential for ASH2L recruitment to the inactive X and regulates histone modifications and escape gene expression. PLoS Genetics, 13(7), e1006890.
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5

Skeletal Muscle Differentiation of ESCs

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ESCs were cultured as previously described (Shakya et al., 2015a) with 2i conditions: ERK inhibitor PD0325901 (1 μM, LC Laboratories) and GSK3 inhibitor CHIR99021 (3 μM, LC Laboratories). Cultures were maintained on irradiated feeders (ThermoFisher). Prior to all experiments ESCs were plated on gelatin to deplete the feeders. For MD differentiation, ESCs were plated on gelatin and cultured as previously described (Chal et al., 2015) . Briefly, parental and cKO cells were cultured in N2B27 medium supplemented with recombinant Bmp4 (Peprotech) for 2 d. After 48 hr, media was changed to RDL (Rspo3, DMSO, LDN) medium. Cells were harvested 24 hr (day 3) or 96 hr (day 6) later. For muscle differentiation, cells were switched to HIFL (Hgf, Igf, Fgf, Ldn) medium and cultured for 48 hr (day 8) after which medium was switched to 2% horse serum (ThermoFisher). Cells were harvested on day 11 (overexpression experiments) or 19 (RT-qPCR).
Perovanović J., Wu Y., Abewe H., Shen Z., Hughes E.P., Gertz J., Chandrasekharan M.B., & Tantin D. (2020). Oct1 cooperates with Smad transcription factors to promote mesodermal lineage specification.
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6

Feeder-Free Expansion of Embryonic Stem Cells

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ES cells were cultured in ESC media (15% FBS, 25 mM HEPES, 1× GlutaMAX, 1× MEM Non-essential Amino Acids Solution, 1× Penicillin/Streptomycin, and 0.055 mM β-Mercaptoethanol in DMEM High Glucose (4.5 g/L)) containing 2i (1 μM PD0325901, LC Laboratories; and 3μM CHIR99021, LC Laboratories) and LIF (1300 U/mL, in-house) on cell culture plates coated with 0.2% gelatin under feeder-free conditions. HEK293T cells (CRL-11268, ATCC) were cultured in DMEM High Glucose supplemented with solution (10% FBS, 1× Penicillin/Streptomycin, 1 mM sodium pyruvate, 1× MEM Non-essential Amino Acids Solution, and 1× GlutaMAX) on cell culture plates. The expanded ES colonies and confluent HEK293T cells were dissociated using 0.25% trypsin-EDTA solution for passaging.
Sakashita A., Maezawa S., Takahashi K., Alavattam K.G., Yukawa M., Hu Y.C., Kojima S., Parrish N.F., Barski A., Pavlicev M, & Namekawa S.H. (2020). Endogenous retroviruses drive species-specific germline transcriptomes in mammals. Nature structural & molecular biology, 27(10), 967-977.
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7

Inhibitor Regulation of MAPK Signaling

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TAPI-1 and PD0325901 were purchased from Calbiochem (San Diego, CA). PD0325901 and TPA were purchased from LC laboratories (Woburn, MA). PD153035 was purchased from MedChemexpress (Princeton, NJ).
Hiratsuka T., Fujita Y., Naoki H., Aoki K., Kamioka Y, & Matsuda M. (2015). Intercellular propagation of extracellular signal-regulated kinase activation revealed by in vivo imaging of mouse skin. eLife, 4, e05178.
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8

Breast Cancer Cell Line Characterization

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BT549 and SKBR3 cells were obtained from the UCSF Cell Culture Facility.
BT20, BT474, HCC1428, HCC38, LY2, MCF7, MDAMB231, MDAMB453, T47D, SUM52PE, and
ZR75B cell lines were obtained from the American Type Culture Collection (ATCC).
Cell lines used for proteomic profiling and molecular analyses were
authenticated by STR analysis. Lines were grown according to published
protocols50 except for
SKBR3 which was cultured using RPMI media supplemented with 10% fetal
bovine serum (FBS) and 1% pen/strep. All cell lines tested negative for
mycoplasma contamination. Drugs used for cell culture experiments in this study
were purchased from Selleck Chemicals (GDC-0941, MK2206, PD0325901, Lapatinib,
MLN8237, and LEE011) and LC Laboratories (RAD001).
Donnella H.J., Webber J.T., Levin R.S., Camarda R., Momcilovic O., Bayani N., Shah K.N., Korkola J., Shokat K.M., Goga A., Gordan J.D, & Bandyopadhyay S. (2018). Kinome rewiring reveals AURKA limits PI3K-pathway inhibitor efficacy in breast cancer. Nature chemical biology, 14(8), 768-777.
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9

Expansion of Feeder-free Mouse ESCs

Check if the same lab product or an alternative is used in the 5 most similar protocols
ES cells were cultured in ESC media (15% FBS, 25 mM HEPES, 1× GlutaMAX, 1× MEM Non-essential Amino Acids Solution, 1× Penicillin/Streptomycin, and 0.055 mM β-Mercaptoethanol in DMEM High Glucose (4.5 g/L)) containing 2i (1 μM PD0325901, LC Laboratories; and 3μM CHIR99021, LC Laboratories) and LIF (1300 U/mL, in-house) on cell culture plates coated with 0.2% gelatin under feeder-free conditions. The expanded ES colonies were dissociated using 0.25% trypsin-EDTA solution for passaging.
Maezawa S., Sakashita A., Yukawa M., Chen X., Takahashi K., Alavattam K.G., Nakata I., Weirauch M.T., Barski A, & Namekawa S.H. (2020). Super-enhancer switching drives a burst in gene expression at the mitosis-to-meiosis transition. Nature structural & molecular biology, 27(10), 978-988.
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10

Doxycycline-inducible Dux ESC Line

Cited 1 time
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Doxycycline (Dox)-inducible Dux ESC line was generated using the PiggyBac Transposon System in our laboratory (hereafter referred to as ESCDUX), in which Dux expression was induced in the presence of Dox21 (link). The ESCDUX was cultured in ESC medium (10% FBS, 1× sodium pyruvate, 1× GlutaMAX, 1× MEM non-essential amino acids solution, 1× penicillin/streptomycin and 0.055 mM β-mercaptoethanol in DMEM high glucose (4.5 g liter−1)) containing 2i (1 µM PD0325901, LC Laboratories; and 3 µM CHIR99021, LC Laboratories) and LIF (1,000 U ml−1, in-house) on cell-culture plates coated with 0.1% gelatin under feeder-free conditions. The expanded colonies were dissociated using 0.05% trypsin-EDTA solution for passaging.
Sakashita A., Kitano T., Ishizu H., Guo Y., Masuda H., Ariura M., Murano K, & Siomi H. (2023). Transcription of MERVL retrotransposons is required for preimplantation embryo development. Nature Genetics, 55(3), 484-495.
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