Dorsomorphin

Manufactured by Cayman Chemical

Sourced in United States

Dorsomorphin is a chemical compound used as a tool in laboratory research. It functions as a selective inhibitor of bone morphogenetic protein (BMP) signaling pathways. Dorsomorphin can be utilized in experiments to investigate the role of BMP signaling in various biological processes.

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Dorsomorphin (compound C) (2 citations)

13 protocols using dorsomorphin

Tendon Cell Ablation and Regeneration Assays

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For tendon cell ablation, animals at indicated time points were treated with 5mM Mtz (Sigma, M3761) in embryo medium in the dark for 1 or 2 days at 28.5°C. The efficacy of ablation was confirmed by checking the RFP fluorescence. At the end of ablation, the treated animals were fixed for staining and analysis or transferred to petri dishes or the zebrafish housing system to raise for regeneration assays. For Dorsomorphin (Cayman, 866405–64-3) treatments, 6 dpf ablated embryos (1 day post ablation) were incubated in 10uM Dorsomorphin for 3 days. For LDN-193189 (Cayman, 1062368–62-0) treatment, 6 dpf ablated embryos were bathed in 10uM LDN-193189 for 2 days. For Dorsomorphin treatment at 1–3 dpf, 1 dpf embryos were bathed in 20uM Dorsomorphin for 2 days.

Niu X., Subramanian A., Hwang T.H., Schilling T.F, & Galloway J.L. (2020). Tendon cell regeneration is mediated by attachment site-resident progenitors and BMP signaling. Current biology : CB, 30(17), 3277-3292.e5.

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Differentiation of iPSCs into NSCs

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We differentiated iPSCs into NSCs by following a default differentiation protocol as previously described^{9 (link),55 (link)}. Briefly, iPSC colonies were detached by collagenase (Gibco) to form embryoid bodies (EBs) and then cultured in suspension in the presence of 1 μM dorsomorphin (Cayman Chemical). After floating for seven days, EBs were attached on poly-ornithine (10 μg/ml, Life Technologies) and laminin (5 μg/ml, Life Technologies) coated surfaces. At day 16, emerging rosette colonies were manually detached, gently dissociated with accutase (Innovative Cell Technologies) and plated as NSC monolayers onto coated dishes for further neuronal differentiation.

Schafer S.T., Paquola A.C., Stern S., Gosselin D., Ku M., Pena M., Kuret T.J., Liyanage M., Mansour A.A., Jaeger B.N., Marchetto M.C., Glass C.K., Mertens J, & Gage F.H. (2019). Pathological priming causes developmental gene network heterochronicity in autism patient-derived neurons. Nature neuroscience, 22(2), 243-255.

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Anterior Foregut Endoderm Differentiation

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Anterior foregut endoderm was differentiated from definitive endoderm cells and treated for 72 h with anterior foregut endoderm differentiation medium containing Ham’s F-12 Nutrient Mix (Thermo Fisher Scientific, Waltham, MA, USA) and IMDM (Thermo Fisher Scientific) with B27 Supplement (Thermo Fisher Scientific), N2 Supplement (Thermo Fisher Scientific), 0.1% bovine serum albumin fraction V (Millipore Sigma, St. Louis, MO, USA), 1-thioglycerol (Millipore Sigma), 1× GlutaMAX Supplement (Thermo Fisher Scientific), 1% penicillin-streptomycin, 50 μg/mL L-ascorbic acid, 10 mM SB431542 (Cayman Chemical, Ann Arbor, MI, USA), and 2 mM dorsomorphin (Cayman Chemical).

Hirai H., Liang X., Sun Y., Zhang Y., Zhang J., Chen Y.E., Mou H., Zhao Y, & Xu J. (2021). The sodium/glucose cotransporters as potential therapeutic targets for CF lung diseases revealed by human lung organoid swelling assay. Molecular Therapy. Methods & Clinical Development, 24, 11-19.

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Investigating Diazoxide-Induced K-ATP Channel Currents

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Drugs and chemicals were dissolved in water or DMSO as stock solutions before being added to superfusate at 1:1000 dilutions. Control studies showed no effect of this DMSO concentration on membrane conductance or firing rate. Superfusate containing a chemical reached the recording chamber within 1 min after passing through a heat exchanger. The K-ATP channel opener Diazoxide was superfused for 5 min every 20 min; net Diazoxide-induced currents were measured as the difference between peak current and current immediately before application of Diazoxide. Test agents (dorsomorphin, STO-609, U73122, m-3M3FBS, m-CPP, and DHPG) were added to the superfusate 5 min after the first application of Diazoxide. The first application of Diazoxide was begun about 10 min after starting whole-cell recording. In some experiments dorsomorphin was added to the pipette internal solution and allowed to diffuse passively into the cell. Diazoxide was obtained from Sigma-Aldrich (St. Louis, MO, USA), m-CPP (meta- chlorphenylpiperazine) was from Tocris Cookson (Bristol, UK), dorsomorphin and STO-609 were from Cayman Chemical (Ann Arbor, MI, USA), U73122 and m-3M3FBS were from R&D Systems (Minneapolis, MN, USA), and DHPG ((S)-3,5-dihydroxyphenylglycine) was from Ascent Scientific (Cambridge, MA, USA).

Shen K.Z., Munhall A.C, & Johnson S.W. (2018). Phosphoinositol metabolism affects AMP kinase-dependent K-ATP currents in rat substantia nigra dopamine neurons. Brain research, 1706, 32-40.

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Directed Differentiation of iPSCs to Neural Stem Cells

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For embryoid body (EB) formation, jm-iPSCs were dissociated and transferred to 6-well low attachment culture plates (corning) at 1 × 10⁶ cells/ml in 15%FBS/DMEM. After 2 weeks culture, the EBs were transferred to Geltrex-coated plates and cultured at an adhesion condition with 15%FBS/DMEM for another 3 weeks.
For directed differentiation into neural stem cells, jm-iPSCs were transferred to 6-well low attachment plates in KBM Neural Stem Cell medium (Kohjin Bio, 16050200) supplemented with 2 μM dorsomorphin (Cayman, 11967), 10 μM SB431542 (Cayman, 13031), and 1 × B-27 (Gibco, 17504-044) at 1.5 × 10⁵ cells/ml. Soon after iPSC dissociation, 10 μM Y-27632 was added to the medium. On day 1 and 4, 3 ng/ml Stembeads FGF2 (StemCultures, SB500) were added into the medium and the medium was half-exchanged on day 4. To induce neuronal differentiation, 1-week old neurospheres were plated onto Geltrex-coated culture plates and cultured in KBM Neural Stem Cell medium without FGF2 and EGF but supplemented with 1 × B-27 for 2 weeks.

Nakai R., Ohnuki M., Kuroki K., Ito H., Hirai H., Kitajima R., Fujimoto T., Nakagawa M., Enard W, & Imamura M. (2018). Derivation of induced pluripotent stem cells in Japanese macaque (Macaca fuscata). Scientific Reports, 8, 12187.

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Directed Differentiation of hESCs to NPCs

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The cells were grown on MEFs to similarly sized, defined colonies using hESC medium. Induced embryoid body (EB) formation was achieved by detaching colonies with collagenase IV for 30 min. The cells were centrifuged and transferred as colonies to a Petri dish containing hESC medium without basic fibroblast growth factor (bFGF) + 5 μM dorsomorphin (catalog #11967, Cayman) and 5 μM SB431542 (SB #13031, Cayman). The EBs were cultured for 4–5 days with medium replacement every other day. For the NPC expansion, the cells were plated on Matrigel-coated plates (without dissociation) and cultured with KO DMEM medium + 1XN2 supplement (stock X100) and 20 ng/mL bFGF for 8–10 days with medium replacement every other day.

Cohen-Hadad Y., Altarescu G., Eldar-Geva T., Levi-Lahad E., Zhang M., Rogaeva E., Gotkine M., Bartok O., Ashwal-Fluss R., Kadener S., Epsztejn-Litman S, & Eiges R. (2016). Marked Differences in C9orf72 Methylation Status and Isoform Expression between C9/ALS Human Embryonic and Induced Pluripotent Stem Cells. Stem Cell Reports, 7(5), 927-940.

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Directed Neuronal Differentiation of iPSCs

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The neuronal differentiation of iPSCs was performed as previously described [80 (link)]. Briefly, iPSCs were dissociated using Accutase (FoliBio, Taiwan) and seeded on Matrigel-coated plates supplemented with StemFlex medium. The cells were then cultured in 3 N medium (DMEM/F12, neurobasal medium, non-essential amino acids, sodium pyruvate, Glutamax; penicillin-streptomycin, B27, N2, 2-mercaptoethanol, all from Thermo Fisher Scientific, MA, USA) containing 10 μM SB431542 (Selleckchem, TX, USA) and 1 μM Dorsomorphin (Cayman Chemical, MI, USA) for 12 days. The formed neuroepithelial clumps were digested by dispase, reseeded on Matrigel-coated plates, and incubated in 3 N medium containing 20 ng/ml bFGF (Thermo Fisher Scientific, MA, USA) for four days. Next, the cells were maintained in a 3 N medium without bFGF, which was replaced every three days until used for the experiments. After four weeks of differentiation, TUJ1 expression was detected by flow cytometry (FACS Calibur, BD Biosciences, NJ, USA). Neuronal differentiation for more than four weeks was also performed using a 3 N medium without bFGF.

Chen T.Y., Lin S.P., Huang D.F., Huang H.S., Tsai F.C., Lee L.J., Lin H.Y, & Huang H.P. (2024). Mature neurons from iPSCs unveil neurodegeneration-related pathways in mucopolysaccharidosis type II: GSK-3β inhibition for therapeutic potential. Cell Death & Disease, 15(4), 302.

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Exploring Ferroptosis Regulators in Cells

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Regents used in this study including Rotenone (no. S2348, Selleck), TW-37 (no. 20999, Cayman Chemical), Rapamycin (no. HY-10219, MCE),Dorsomorphin (no. 11967, Cayman Chemical), 5-Fluorouracil (no. HY-90006, MCE), Shikonin (no. HY-N0822, MCE), FIN56 (no. HY-103087, MCE), Polyphyllin VI (no. S9302, Selleck), RSL3 (no. 19288, Cayman Chemical), Z-VAD-FMK (no. HY-16658B, MCE), Necrostatin-1 (no. HY-15760, MCE), Ferrostatin-1 (no. HY-100579, MCE), Erastin (no. S7247, Selleck), iFSP1 (no. 29483, Cayman Chemical), Sorafenib (no. 10009644, Cayman Chemical), BSO (no. S9728, Selleck), Acetylcysteine (no. HY-B0215, MCE), GSH (no. HY-D0187, MCE), Cisplatin (no. HY-17394, MCE), Abrine (no. HY-N1436, MCE) were obtained from the indicated vendors.
The antibodies were as follows: SPTBN2 (Abcam, no. ab238055 for IF, IHC, WB; no.ab264178 for IP), SLC7A11 (Proteintech, 26864-1-AP for WB; Abcam, no.ab307601 for IF, IHC, WB, IP, Flow Cyt), Arp1 (Abcam, no.ab203833), Flag (Abcam, no.ab205606), HA (Abcam, no.ab9110); Tubulin (Proteintech, no.11224-1-AP), Vinculin (Proteintech, no.66305-1-Ig), Na⁺/K⁺ATPase (Beyotime, no.AF1864).

Deng J., Lin X., Qin J., Li Q., Zhang Y., Zhang Q., Ji C., Shen S., Li Y., Zhang B, & Lin N. (2024). SPTBN2 suppresses ferroptosis in NSCLC cells by facilitating SLC7A11 membrane trafficking and localization. Redox Biology, 70, 103039.

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Differentiation of iPSCs into NSCs

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Inhibition of EHEC Infection in BMDMs

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BMDMs were pretreated with 10 μM of dorsomorphin (11967, Cayman Chemicals) or 10 μM of kb-NB 142-70 (18002, Cayman Chemicals) 1 h prior to infection with EHEC or ΔEhaF at MOI = 50. At 1 h of infection, media was replaced with gentamicin containing media containing the corresponding inhibitors. Cell lysates and supernatants were collected at indicated time points to assess the levels of TFE3 and cytokines.

Ta A., Ricci-Azevedo R., Vasudevan S.O., Wright S.S., Kumari P., Havira M.S., Surendran Nair M., Rathinam V.A, & Vanaja S.K. (2023). A bacterial autotransporter impairs innate immune responses by targeting the transcription factor TFE3. Nature Communications, 14, 2035.

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