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Vitamin d2

Manufactured by Santa Cruz Biotechnology
Sourced in United States, Germany

Vitamin D2 is a laboratory product used for research purposes. It is a form of vitamin D that is derived from plant sources. Vitamin D2 serves as a precursor for the production of active vitamin D metabolites, which are essential for various biological processes.

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2 protocols using vitamin d2

1

Vitamin D Analogs in A549 Cells

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Alveolar epithelial cells type II (A549, ATCC) were maintained in DMEM medium supplemented with 10% FBS (Sigma-Aldrich, Burlington, MA, USA), 2 mM glutamine and 100 U/mL of penicillin and streptomycin (Lonza, Basel, Switzerland). We used the active form of vitamin D (1α,25-Dihydroxyvitamin D3 or calcitriol) (cat.#D1530; Sigma-Aldrich; Vitamin D stock was 10 μM in ethanol) and the following vitamin D analogs (stocks were 50 μM in ethanol): calcipotriol (cat.#203537; Santa Cruz Biotechnology, Dallas, TX, USA), paricalcitol (cat.#477938; Santa Cruz Biotechnology), tacalcitol (cat.#sc-361371a; Santa Cruz Biotechnology, Dallas, TX, USA), 22-Oxacalcitriol (cat.#sc-361076; Santa Cruz Biotechnology, Dallas, TX, USA) and vitamin D2 (cat.#sc- sc-205988; Santa Cruz Biotechnology, Dallas, TX, USA). Treatments were performed in cells maintained in DMEM supplemented with 10% hormone-depleted serum. This serum was prepared by using the anion exchange resin AGR1-X8 from BIO-RAD (cat.#1401441; BIO-RAD, Hercules, CA, USA) as previously described [20 (link)]. Bleomycin sulfate (cat.# CAYM13877–50) was purchased to VWR [Radnor, PA, USA] (bleomycin stock: 50 mM in PBS).
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2

Quantifying Cell Proliferation using BrdU Assay

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DNA synthesis was quantified employing the 5-bromo-2’-deoxyuridine (BrdU) labelling and detection enzyme-linked immunosorbent assay kit (Roche Diagnostics, Mannheim, Germany). Therefore, quiescent or activated (proliferating) PSCs were plated in 96-well plates at equal seeding densities and allowed to adhere overnight. Afterwards, the cells were exposed to vitamin D2, vitamin D3 (both from Santa Cruz Biotechnologies, Heidelberg, Germany) or calcipotriol (Sigma-Aldrich, Deisenhofen, Germany) for the indicated periods of time. Twenty-four hours prior to cell harvesting, BrdU labelling was initiated by adding labelling solution at a final concentration of 10 μmol/L. Afterwards, labelling was stopped, and BrdU uptake was measured according to the manufacturer’s instructions.
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