Naive CD4+ T cells were plated under iTreg differentiation conditions at 1.0x105 cells/well in 96 U-bottom well plates (Falcon). For cell stimulation, plates were coated at least 5 hours prior to use with 5 ug/ml plate-bound anti-CD3 antibody (clone OKT3; BioLegend, LEAF grade), which is then washed off with PBS, with 1 ug/ml soluble anti-CD28 antibody (BioLegend, LEAF grade) and 5 ng/ml IL-2 (carrier-free; Tocris R&D Systems) added with cells. Cells treated with only these functioned as the mock control. For iTreg induction, TGF- β (5ng/ml; carrier-free; Tocris R&D Systems), anti-IFN γ antibody (2 ug/ml; clone MD-1, Biolegend), bexarotene (1 µM, Abcam) or atRA (100 nM; Sigma-Aldrich) were added additionally. The DMSO control (for bexarotene, atRA and AGN190) had no effect on FOXP3 expression. Cells were incubated for 7 days unless otherwise stated.
Bexarotene
Manufactured by Abcam
Sourced in United Kingdom
Bexarotene is a synthetic retinoid compound. It is used as a laboratory reagent for research purposes.
Automatically generated - may contain errors
Lab products found in correlation
Soluble anti cd28 antibody, by BioLegend (2 mentions)
96 u bottom well plates, by BD (2 mentions)
Clone md 1, by BioLegend (2 mentions)
Anti cd3 antibody clone okt3, by BioLegend (2 mentions)
Tgf β, by R&D Systems (2 mentions)
Interleukin 2 (il 2), by R&D Systems (2 mentions)
Il 1β, by R&D Systems (1 mentions)
Interleukin 6 (il 6), by R&D Systems (1 mentions)
Il 23, by R&D Systems (1 mentions)
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
β actin, by Proteintech (1 mentions)
Pcdna3, by Genecreate (1 mentions)
3 protocols using bexarotene
1
Induction of Regulatory T Cells
2
Th17 Cell Differentiation Protocol
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After either MACS or FACS isolation, naive cells were plated under Th17 differentiation conditions at 5x105 cells/well in 96 U-bottom well plates (Falcon). For cell stimulation, plates were coated at least 5 hours prior to use with 3 ug/ml plate-bound anti-CD3 antibody (clone OKT3; BioLegend, LEAF grade), 1 ug/ml soluble anti-CD28 antibody (BioLegend, LEAF grade) and 2 ng/ml IL-2 (carrier-free; Tocris R&D Systems). Cells treated with only these functioned as the mock control. For Th17 induction, TGF- β (30 ng/ml; carrier-free; Tocris R&D Systems), IL-6 (30 ng/ml; carrier-free; Tocris R&D Systems), IL-1 β (10 ng/ml; carrier-free; Tocris R&D Systems), IL-23 (50 ng/ml; carrier-free; Tocris R&D Systems), anti-IFN γ antibody (2 ug/ml; clone MD-1, Biolegend), anti-IL-4 antibody (2 ug/ml; clone MD-1, Biolegend), bexarotene (1 µM, Abcam), or atRA (100 nM; Sigma-Aldrich) were added additionally. The DMSO control (for bexarotene, atRA and AGN190) had no effect on IL-17A cytokine expression. Cells were incubated for 7 days unless otherwise stated.
3
Bexarotene Neuroprotection in HT22 Cells
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Bexarotene was purchased from Abcam Bio-tech （Cambridge, MA, UK). The immortalized mouse hippocampal neuron line (HT22) was obtained from JENNIO Biological Technology (Guangzhou, China).The sodium dodecyl sulfate (SDS), Acr-Bis, Bovine serum albumin (BSA) were purchased from GEN-VIEW SCIENTIFIC Inc (Florida, USA). The Tunelkit , Phosphate Buffer Saline (PBS) , Tris-HCl buffer, Methyl Thiazolyl Tetrazolium (MTT), Fetal bovine serum (FBS), Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12(DMEMF-12) and Dulbecco’s modified Eagle’s medium (DMEM) were purchased from Dingguo Co. Ltd. (Beijing, China). Antibodies against JIP1, JIP3, phospho-JNK (1/2), phospho-ASK1, ASK1, Cleaved Caspase 3 were purchased from Cell Signaling Technology (Danvers, MA, USA). Antibodies against β-actin and horse radish peroxide (HRP)-conjugated goat anti-rabbit IgG (H + L) were purchased from Proteintech Biotechnology (Proteintech, Wuhan, China). The plasimid vector of pCDNA 3.1 was purchased from Gene Create Biological Engineering (Wuhan, China).
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