Fluka silica gel

Ascorbic acid-2-phosphate, β-glycerophosphate, dexamethasone, 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT), P-nitrophenyl phosphate (pNPP), Alizarin Red S (AR-S), paraformaldehyde, Triton X-100, fetal calf serum (FCS), L-glutamine and antibiotics (penicillin and streptomycin) were purchased from Sigma-Aldrich (Saint Louis, MO, USA). Alexa Fluor 488 Phalloidin (#A12379) was purchased from Thermo Fisher Scientific (Waltham, MA, USA). Antibody against human Runx2 (#sc-10758) was purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Dulbecco’s modified eagle’s medium high glucose (DMEM), Ham’s F12 and phosphate-buffered saline (PBS 1X) were purchased from Euroclone (Milan, Italy). All organic solvents, HPLC-grade (methanol, acetone, n-hexane, dichloromethane), were supplied from Carlo Erba Reagents (Dasit Group Carlo Erba Reagenti, Milan, Italy). TLC plates (Analtech Uniplate, Silica gel GF, 20 × 20 cm, 500 microns) for preparative chromatography were purchased from Merck Sigma-Aldrich (Merck Sigma-Aldrich, Milan, Italy). Cromatographic column was performed on silica gel using column chromatography (Merck Sigma-Aldrich 60, 70-230 mesh, Fluka^® silica gel).

Solvents were
purified by conventional
methods, distilled, and stored over activated molecular sieves under
argon. All the chemicals were purchased from commercial sources and
used as received without purification. All the operations under an
inert atmosphere were carried out using standard Schlenk techniques
and employing dried nitrogen. Reactions that required heating were
performed in an oil bath. For all reactions, conversion was monitored
by thin-layer chromatography analysis on pre-coated silica gel plates
(VWR Macherey-Nagel, 0.2 mm thick) or pre-coated neutral alumina plates
(Sigma-Aldrich, 0.25 mm thick). Column chromatography was performed
with Fluka silica gel, pore size 60 c5, 70–230 mesh, 63–200
μm or Sigma-Aldrich activated, neutral alumina. ¹H NMR and ¹³C NMR spectra were recorded at room temperature
in CDCl₃ or DMSO-d₆ solution
with a Varian INOVA-600 spectrometer, operating at a frequency of
600 MHz for ¹H and 150 MHz for ¹³C, using the
residual solvent peak as the internal reference; chemical shift (δ)
values are given in parts per million (ppm) and coupling constants
(J) in Hz. Mass spectra were obtained with an Applied
Biosystems-MDS Sciex API 4000 triple quadrupole mass spectrometer
(Concord, Ont., Canada), equipped with a Turbo-V ion-spray (TIS) source.
Elemental analyses were performed on a Elementar Vario Micro Cube
CHN-analyzer.