hpDPSCs and MDPSCs were stimulated with IFN-γ (PROSPEC, East Brunswick NJ, USA) at a concentration of 20 ng/ml in DMEM without serum for 24 h according to the previous studies [27 (link), 28 (link)] with slight modification. Non-stimulated cells were used as a control. RNA was extracted using Trizol, and the CM was collected and concentrated. The mRNA expression of immunosuppressive markers IDO, TGF-β1, PTGE, and IL-6 as our previous study [29 (link)] was examined by RT-PCR. The concentration of nitric oxide (NO) was examined by measuring its stable end product, nitrite, in the CM using a Griess reagent (Promega Corporation, Madison, WI, USA) according to manufacturer’s protocol. Absorbance at 540 nm was measured by microplate reader (SpectraMax Gemini XPS/EM, Molecular Devices, San Jose, CA, USA), and nitrite concentrations were calculated using a standard nitrite curve.
Spectramax gemini xps em
Manufactured by Molecular Devices
Sourced in United States
The SpectraMax Gemini XPS/EM is a multimode microplate reader that can perform absorbance, fluorescence, and luminescence measurements. It is designed to provide accurate and reliable data for a wide range of assays and applications.
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Lab products found in correlation
2 protocols using spectramax gemini xps em
1
Immunosuppressive Marker Expression in hpDPSCs and MDPSCs
2
Heterologous Expression of VlAbcG1a in Yeast
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To identify the homologs of VlAbcG1a and VlAbcG1b in S. cerevisiae, phylogenetic analysis was conducted as described above. For heterologous expression in WT and deletion strains, the VlAbcG1a gene was amplified using cDNA from V. longisporum and cloned into the pYES‐2 vector driven by the GAL1 promoter, while the empty pYES‐2 vector was used as a negative control, followed by transformation using a polyethylene glycol‐based protocol in S. cerevisiae BY4742 (Gietz & Schiestl, 2007 (link)). The positive transformants were selected on synthetic complete (SC) without uracil (−URA) medium. For gene induction, strains were precultured in SC–URA medium with 1% raffinose to reach the log phase. Then, the OD600 was adjusted to 0.3 and transferred to SC–URA medium supplemented with 2% galactose. The growth of strains was investigated on exposure to 0.003% β‐pinene and measuring the OD600 in SpectraMax Gemini XPS/EM microplate reader (Molecular Devices) at 30°C in a time‐course assay. Five replicates per treatment were used.
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