To evaluate the effect of EGCG on cell growth of DPCs and ORSCs, DPCs and ORSCs were treated with different concentrations of EGCG (0.25, 0.5, 1, 2, and 4 μM) for 12, 24, or 48 h. To investigate whether the Shh and AKT signaling pathways are involved in the effect of EGCG on growth of DPCs and ORSCs, the cells were divided into the following groups, treated accordingly and incubated for 48 h: (1) Control; (2) EGCG (0.5 μM); (3) Cyclopamine (5 μM, Medchem Express, Monmouth Junction, NJ, United States); (4) EGCG+Cyclopamine; (5) GANT61 (10 μM; Medchem Express); (6) EGCG+GANT61; (7) LY2940002 (10 μM; Beyotime); (8) EGCG+LY2940002.
Cyclopamine
Manufactured by MedChemExpress
Sourced in United States
Cyclopamine is a chemical compound that acts as a hedgehog pathway inhibitor. It is commonly used in research settings to study cellular signaling pathways and developmental processes.
Automatically generated - may contain errors
Lab products found in correlation
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (4 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Gant61, by MedChemExpress (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin solution, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by MedChemExpress (1 mentions)
Ly2940002, by Beyotime (1 mentions)
Nerve growth factor (ngf), by Thermo Fisher Scientific (1 mentions)
Sh sy5y, by Merck Group (1 mentions)
Horse serum, by Merck Group (1 mentions)
6 ohda, by Merck Group (1 mentions)
Sh sy5y, by Procell (1 mentions)
A549 cells, by Thermo Fisher Scientific (1 mentions)
Mouse anti crkl, by Santa Cruz Biotechnology (1 mentions)
Mouse anti flag tag, by Merck Group (1 mentions)
Adriamycin, by Merck Group (1 mentions)
F 12k medium, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Cisplatin, by Merck Group (1 mentions)
Penicillin streptomycin, by Sangon (1 mentions)
5 protocols using cyclopamine
1
EGCG's Impact on DPC and ORSC Growth
2
Inhibition of Sonic Hedgehog Pathway in Ovarian Granulosa Cells
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The human ovarian granulosa cell line, KGN (Procell, Wuhan, China), was cultured in Dulbecco’s Modified Eagle Medium (DMEM; Invitrogen, Carlsbad, CA, USA) containing 10% fetal bovine serum (FBS; Hyclone, Logan, UT, USA) and 1% penicillin-streptomycin solution (Invitrogen) at 37 °C. To inhibit the sonic hedgehog (SHH) signalling pathway, 5 μM cyclopamine (MedChemExpress, Monmouth Junction, NJ, USA), an inhibitor of the SHH signalling pathway, was used to treat KGN cells for 24 h [22 (link)].
3
Osteoblast Behavior on Titanium Surfaces
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The human MG63 osteoblasts were obtained from ATCC (Rockville, MD, USA). The human MG63 osteoblasts were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM; Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 1% antibiotic mixture (penicillin/streptomycin; HyClone, Logan, UT, USA) and 10% fetal bovine serum (FBS; Thermo Fisher Scientific) at 37°C under a 5% CO2 humidified atmosphere. The MG63 osteoblasts were seeded onto the four different surfaces of titanium disks in 12-well plates with a concentration of 5×104/well. The cyclopamine (MedChem Express, Princeton, NJ, USA) was dissolved at 5 mg/mL in dimethyl sulphoxide, and the solution was diluted to the final concentration with DMEM supplemented with 1% antibiotic mixture (penicillin/streptomycin) and 10% FBS. Treatment with control vehicle was used for control cells.
4
Neurotoxin Exposure in PC-12 and SH-SY5Y Cells
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The rat adrenal pheochromocytoma cell line PC-12 and the human neuroblastoma cell line SH-SY5Y were purchased from Procell (Wuhan, China). PC-12 cells were cultured in RPMI-1640 medium (Gibco, Grand Island, NY, USA) supplemented with 10% horse serum (Sigma, St. Louis, MO, USA) and 5% fetal bovine serum (FBS; Hyclone, Logan, UT, USA). SH-SY5Y cells were cultured in RPMI-1640 medium supplemented with 10% FBS. Cells were maintained at 37°C in an atmosphere of 95% air and 5% CO 2 . Highly differentiated PC-12 cells were induced using 50 ng/mL nerve growth factor (Gibco) for 4 days.
For neurotoxin exposure, PC-12 and SH-SY5Y cells were exposed to 1, 5, 10, 50, or 100 μm 6-OHDA (Sigma) for 24 h followed by two washes with phosphate-buffered saline and were used in the following experiments. To inhibit the SHH signaling pathway, cells were treated with 5 μm cyclopamine (MedChemExpress, Monmouth Junction, NJ, USA) for 24 h.
For neurotoxin exposure, PC-12 and SH-SY5Y cells were exposed to 1, 5, 10, 50, or 100 μm 6-OHDA (Sigma) for 24 h followed by two washes with phosphate-buffered saline and were used in the following experiments. To inhibit the SHH signaling pathway, cells were treated with 5 μm cyclopamine (MedChemExpress, Monmouth Junction, NJ, USA) for 24 h.
5
Evaluating Anti-Cancer Drug Effects
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All cell lines used in this study were purchased from ATCC and cultured as following: A549 cells with F‐12K medium (Gibco) containing 10% FBS (Gibco) and 1% penicillin/streptomycin (Sangon Biotech), H1299, H1975 and H1650 with RPMI‐1640 medium (Gibco) containing 10% FBS and 1% penicillin/streptomycin, 16HBE cells with DMEM (Gibco) medium containing 10% FBS and 1% penicillin/streptomycin. Cell transfection with plasmids or siRNAs was carried out as previously described.17 The antibodies used for Western blot were as follows: mouse anti‐CRKL (1:1000; Santa Cruz); mouse anti‐Flag tag (abbreviation for Fg; 1:5000; Sigma); rabbit anti‐GLI2 (1:1000; ABclonal); and mouse anti‐ACTIN (1:5000; Genscript). The siRNAs sequences were shown as follows: MOCK‐siRNA, 5′‐CAA ACA CUU CCU UGG AAU GdTdT‐3′; CRKL‐siRNA‐1, 5′‐GCU CUG CUC UAC CAU GUU UdTdT‐3′; CRKL‐siRNA‐2, 5′‐CGT GAA AGU CAC AAG GAU GdTdT‐3′; GLI2‐siRNA‐1, 5′‐GUU CCU CAC GGC GUC GUA GdTdT‐3′; GLI2‐siRNA‐2, 5′‐CAA GAC CGA GCC UGA GGG CdTdT‐3′. For chemical treatment, cells were seeded into 96‐well plate at 1 × 104/well. After 24 hours, cells were treated with 5 μg/mL adriamycin (Sigma), 20 μg/mL 5‐FU (Sigma), 10 μg/mL cisplatin (Sigma), 20 μmol/L cyclopamine (MedChemExpress), 10 μmol/L gefitinib (MedChemExpress) or 10 μmol/L GANT61 (MedChemExpress) for additional 24 hours.
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