Paroxetine was purchased from Apexbio (Suzhou, China). Alpha-MEM and DMEM were procured from Biosharp (Anhui, China). The IL-1β and Cell Counting Kit-8 (CCK-8) were purchased from Med Chem Express (New Jersey, USA). RANKL and M-CSF were procured from LifeTein (Beijing, China). Primary antibodies against Nlrp3 (ab263899, 1:1000), IL-1β (ab254360, 1:1000), Caspase1 (ab179515, 1:1000) and ADAMTS5 (ab185722, 1:1000) were purchased from Abcam (Cambridge, UK). The aggrecan antibody (PA1-1746, 1:1000) was procured from Thermo Fisher Scientific (Waltham, MA, USA), and MMP3 (ET1705-98, 1:1000) and SOX9 (ET1611-56, 1:1000) antibodies were purchased from Huaan Biotechnology (Hangzhou, China). Primary antibodies against P65 (8242, 1:1000), p-P65 (3033, 1:1000), IκBα (4812, 1:1000), p-IκBα (2859, 1:1000), and β-actin (4970, 1:10,000) were procured from Cell Signaling Technology (Danvers, MA, USA). The NFATc1 antibody (YT5381, 1:1000) was procured from ImmunoWay (Shanghai, China).
M csf
Manufactured by LifeTein
Sourced in China
M-CSF is a lab equipment product that functions as a macrophage colony-stimulating factor. It is a cytokine that plays a role in the differentiation and proliferation of macrophages, which are a type of immune cell.
Automatically generated - may contain errors
Lab products found in correlation
Rankl, by LifeTein (2 mentions)
Ab254360, by Abcam (1 mentions)
Ab179515, by Abcam (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Ab185722, by Abcam (1 mentions)
Il 1β, by MedChemExpress (1 mentions)
Dulbecco modified eagle medium (dmem), by Biosharp (1 mentions)
Iκbα 4812, by Cell Signaling Technology (1 mentions)
P iκbα 2859, by Cell Signaling Technology (1 mentions)
Cell counting kit 8 (cck8), by MedChemExpress (1 mentions)
Ab263899, by Abcam (1 mentions)
Enzyme labeling instrument, by Thermo Fisher Scientific (1 mentions)
3 protocols using m csf
1
Paroxetine Modulates RANKL-Induced Osteoclastogenesis
2
Cytotoxicity of FTY720 on Bone Marrow Macrophages
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The cytotoxic effect of FTY720 (MCE, China) on BMMs was studied using the cell counting kit-8 (CCK-8) assay (MCE, USA) according to the manufacturer's protocols. Briefly, BMMs were seeded in 96-well plates at a density of 3 × 103 cells/well in α-MEM containing 25 ng/ml M-CSF (LifeTein, LLC, Beijing, China). After 24 h of incubation, the cells were administered FTY720 (0, 4, 8, 16, 32, 64, 128, 256, and 512 nM) for 48 h, 72 h, and 96 h. At the specified time point, 10 μL of CCK-8 reagent was added to each well. The plate was incubated at 37°C for another 2 hours in the dark. An enzyme labeling instrument (Thermo Fisher Scientific, USA) was used to determine the absorbance at 450 nm.
3
CDDO-Me Inhibits RANKL-Induced Osteoclastogenesis
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BMMs were seeded in 96-well plates at a concentration of 8 × 103 cells per well in α-MEM containing 25 ng/mL M-CSF (LifeTein, LLC., Beijing, China) overnight. The BMMs were stimulated with 100 ng/mL RANKL (LifeTein, LLC., Beijing, China) after adherence. Moreover, various concentrations of CDDO-Me (0 nM, 3.9 nM, 7.8 nM, 15.6 nM, 31.2 nM, 62.5 nM) were added to the cultured cells. The media, which contained 25 ng/mL M-CSF and 100 ng/mL RANKL, were replaced every 2 days, Furthermore, media containing CDDO-Me was changed every 2 days until the Day 7, when OCs formed. Tartrate-resistant acid phosphatase (TRAP) levels in differentiated OCs were evaluated after the cells were fixed with 4% paraformaldehyde for 1 h. OC differentiation was evaluated by counting the number of TRAP-positive multinucleated cells with 3 or more nuclei.
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