Melanoma cells (CRL-1619, American Tissue Cell Culture) were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% heat inactivated fetal bovine serum and 1% penicillin/streptomycin (Life Technologies, Sigma) [22 (link),26 (link),28 (link),46 (link),47 (link)]. Melanoma cells were incubated with 0, 0.0002, 0.002, 0.02 or 0.2 µM 1α,25–dihydroxyvitamin D3 (D1530, Sigma, dissolved in DMSO to a 20 mM stock solution) in DMEM supplemented with 3% heat inactivated fetal bovine serum for 24 h for experiments, to measure cell viability, oxidative DNA/RNA damage, membrane damage, and expression of p53, superoxide dismutase (SOD), interleukin-1 (IL-1), tumor necrosis factor-α (TNF-α), transforming growth factor-β (TGF-β), vascular endothelial growth factor (VEGF), matrix metalloproteinases (MMP)-1 and MMP-2 at the protein and/or promoter levels. The cells were incubated with 0 or 0.02 uM vitamin D to measure the expression of IL-1, TNF-α, TGF-β, VEGF, MMP-1, and MMP-2 at the mRNA levels
D1530
Manufactured by Merck Group
Sourced in United States
The D1530 is a precision laboratory instrument designed for a range of analytical applications. It features high accuracy and repeatability to support reliable data collection. The core function of the D1530 is to perform measurements and analyses, though its specific intended use is not provided.
Automatically generated - may contain errors
Lab products found in correlation
A8960, by Merck Group (3 mentions)
L ascorbic acid 2 phosphate, by Merck Group (3 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
D4902, by Merck Group (2 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (2 mentions)
Glutamax, by Thermo Fisher Scientific (2 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (2 mentions)
Matrigel, by Thermo Fisher Scientific (2 mentions)
2 mercaptoethanol, by Merck Group (2 mentions)
Pz0162, by Merck Group (2 mentions)
Knockout d mem based medium, by Thermo Fisher Scientific (2 mentions)
Ko ksr, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Pt 2501, by Lonza (2 mentions)
D2915, by Merck Group (2 mentions)
β glycerophosphate, by Merck Group (2 mentions)
Bm mscs, by American Type Culture Collection (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
20 protocols using d1530
1
Vitamin D Modulates Melanoma Cell Responses
2
BM-MSCs Osteogenic Differentiation and Autophagy
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human bone marrow mesenchymal stem cells (BM-MSCs; catalog no. PCS-500-012) were obtained from American Type Culture Collection (ATCC; Manassas, VA, USA) and cultured in low-glucose Dulbecco’s modified Eagle’s medium (DMEM; D5030, Sigma-Aldrich, St Louis, MO, USA) supplemented with 10% fetal bovine serum (FBS; F2442; Sigma-Aldrich, St Louis, MO, USA), 100 U/mL Penicillin (Invitrogen, USA), and 100 μg/mL Streptomycin (Invitrogen, USA).
For osteogenic differentiation, the BM-MSCs were cultured in DMEM (Invitrogen, USA) containing 1% FBS (F2442; Sigma-Aldrich, USA), 200 μMl -glutamine (G7513; Sigma-Aldrich, USA), 10 nM dihydroxyvitamin D3 (D1530; Sigma-Aldrich, USA), 10 mM β-glycerolphosphate (50020; Sigma-Aldrich, USA), 100 nM dexamethasone (D4902; Sigma-Aldrich, USA), and 80 μg/mL ascorbic acid phosphate (A8960; Sigma-Aldrich, USA). And the medium was refreshed every 3–4 days.
To evaluate the effects of Forkhead Box O3 (FOXO3) on the autophagy of BM-MSCs, 100 nM autophagy activator Rapamycin (RAPA; R0395; Sigma-Aldrich, USA) was added into the BM-MSCs transfected with lentivirus carrier of small interfering RNA for FOXO3 (siFOXO3). 5 mmol/L autophagy inhibitor 3-methyladenine (3-MA; M9281; Sigma-Aldrich, USA) was added into BM-MSCs transfected with lentivirus carrier of overexpressed FOXO3 plasmid.
For osteogenic differentiation, the BM-MSCs were cultured in DMEM (Invitrogen, USA) containing 1% FBS (F2442; Sigma-Aldrich, USA), 200 μM
To evaluate the effects of Forkhead Box O3 (FOXO3) on the autophagy of BM-MSCs, 100 nM autophagy activator Rapamycin (RAPA; R0395; Sigma-Aldrich, USA) was added into the BM-MSCs transfected with lentivirus carrier of small interfering RNA for FOXO3 (siFOXO3). 5 mmol/L autophagy inhibitor 3-methyladenine (3-MA; M9281; Sigma-Aldrich, USA) was added into BM-MSCs transfected with lentivirus carrier of overexpressed FOXO3 plasmid.
3
Manipulation of Mll1 in Murine ESCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All ESC lines in this study were adapted to the KnockOut D-MEM-based medium (Thermo Fisher Scientific) supplemented with 2 mM Glutamax (Thermo Fisher Scientific), 1X nonessential amino acids (Thermo Fisher Scientific), 1X sodium pyruvate (Thermo Fisher Scientific), 0.1 mM 2-mercaptoethanol (Sigma-Aldrich), 103 U ml−1 LIF (Millipore, ESG1107). Cells were cultured with either 20% FBS (Atlas Biologicals, catalog number: F-0500-D) if grown on top of MEF feeder cells or 15% KO-KSR (Thermo Fisher Scientific 10828028) and 5% FBS if grown on Matrigel (Thermo Fisher Scientific, A1413202) coated plates, or 20% KO-KSR if grown under 2i condition. 2i consists of 1µM PD0325901 (Sigma Aldrich, PZ0162) and 3µM CHIR99021 (Sigma Aldrich, SML1046). 1α,25-Dihydroxyvitamin D3 (Sigma Aldrich, D1530) is used at a final concentration of 1 µM. Mll1 was deleted by treating Mll1fl/fl-Cre-ERTM ESCs with 5µM 4-OHT for 48hrs. MLL1 inhibition was done by MM-401 treatment at a final concentration of 100 µM (Zhang et al., 2016b (link)).
4
Vitamin D Treatment on H1299 NSCLC Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The human NSCLC cell line H1299 was purchased from Chinese Academy of Sciences Cell Bank (Beijing, China). These cells were authenticated by short tandem repeat method. The H1299 cells were cultured in RPMI-1640 medium (CORNING, 10–040-CVR, USA), which contained 10% fetal bovine serum (GIBCO, USA) and 1% penicillin/streptomycin (Shanghai, China) in a 5% CO2 incubator (3111, Thermo) at 37 °C. H1299 cells were treated with 1 × 10–6 mol/L of vitamin D (D1530, Sigma) for 96 h.
5
Fibroblast Monolayer Wound Healing Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Standardized wounds in the fibroblast monolayer were made by a single scraping with a disposable pipette tip, as previously reported [22 (link)], and medium with or without inactivated fetal bovine serum (iFBS) containing calcitriol 100 nM (D1530, Sigma) or vehicle was added. Then fibroblast photos were taken at different time points. In all cases, the wounded area was determined (ImageJ; National Institutes of Health, Bethesda, MD, USA) from 3 representative photographs taken of each well at 0, 24, and 48 h. Results were expressed as the percentage of the wound at each time point for the maximal wounded area (time 0, 100%). These experiments were performed using an Olympus IX81 (Hamburg, Germany) fluorescence inverted microscope, and the Cell^R software v.2.8 was employed to take images manually.
6
Vitamin D Modulates Endothelial Inflammation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
At confluence, cells were starved in serum-free medium for 24 h enriched with VitD (1a,25-dihydroxyvitamin D3, Catalog Number D1530, Sigma-Aldrich, St. Louis, MO, USA; dissolved in ethanol 0.1% following manufacturer’s instruction), at final concentration of 10 nM for 1 h, as already reported [18 (link),19 (link)], and then incubated with native IL-6 (0.5 ng/mL). LPS-stimulated cells (50 µg/mL) served as positive control and non-VitD treated cells served as negative control. Evaluation of Tissue Factor expression was assessed as previously reported [19 (link)] at 1 and 2 h for gene and 6 and 12 h for protein levels. Surface expression and activity were assessed at 6 h. Similarly, ACE2r, Caspase-1 (as main part of the Inflammasome) and CAMs expression were evaluated. Finally, to verify the effective involvement of VitD receptor (VDR) in the action mechanism of VitD, HUVECs were also treated with VDR antagonist ZK159222 (Bayer Schering Pharma AG, Berlin, Germany). This VDR antagonist was used at the same concentration as VitD (10 nM) as already reported [19 (link)]. Expression of TF-mRNA and procoagulant activity were evaluated.
7
HL60 and U937 cell differentiation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HL60 and U937 cells (wild type or transduced) were treated with 50 nM and 30 nM of 1α,25-dihydroxyvitamin D3 (Sigma, D1530), respectively for 72 h to differentiate into monocytes, which was assessed by scoring CD14+ cells (BD Biosciences, 560180) cells using CytoFlex S (Beckman Coulter). Data was analyzed using CytExpert (Beckman Coulter).
8
Vitamin D treatment in Drosophila
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
9
THP-1 Cell Stimulation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The THP-1 cells were collected from T75 flasks (25 mL suspension) and pelleted by centrifugation at 1,500 rpm for 10 min. The experiment involved 4 replicates of untreated cells, 4 replicates of cells treated with 50 nM 1,25-dihydroxyvitamin D3 (Sigma-Aldrich, D1530), 4 replicates of cells treated with 1,000 Hz vibrations (amplitude range of 30 - 60 nm), and 4 replicates of cells treated simultaneously with 50 nM 1,25-dihydroxyvitamin D3 and 1,000 Hz stimulation. The cells underwent stimulation for 3 days (72 h). No medium or vitamin D3 replacement occurred for this duration of time. The cell density per each replicate at the start of the experiment was 1.5 × 105 cells/mL, in one mL suspension plated on 24-well plates (Thermo Fisher Scientific; 142475). The experiments took place at 37 °C, 5% CO2, and 95% air incubator (LEEC 190D CO2).
10
Vitamin D Supplementation in MTX-Induced Bone Damage
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For the vitamin D supplementation trial, MTX treatment was given as described above, and calcitriol (the active form of vitamin D) (D1530, Sigma-Aldrich, St. Louis, MO, USA) (firstly dissolved in 90% ethanol and then diluted in saline before use) was administered subcutaneously once daily at 0.4 mg/kg body weight [26 (link)] both during and after the MTX treatment period until one day before being euthanised. Groups of rats were euthanised on days 6 and 9 after the initial MTX injection (n = 6 rats/group). A group of saline-injected rats was also euthanised on the same day as the MTX-treated day 9 rats, which served as normal controls. Serum and tibial bone specimens were collected [23 (link)], respectively, for biochemical and structural analyses (Figure 1 B). The above protocols followed the Australian Code of Practice for the Care and Use of Animals and were approved by the Animal Ethics Committee of the University of South Australia (approval number: U4-14).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!