Rhohamin phalloidin

All of the reagents, tartaric acid (C₄H₆O₆, Merck Ltd, Mumbai, 99.7%), thioacetamide (C₂H₅NS, S.D. fine chemical Ltd, India, 99%), dopamine hydrochloride (Sigma-Aldrich, 98%), trans-cinnamaldehyde (C₉H₈O, Alfa Aesar, 98%), formaldehyde (CH₂O, Merck, 41% w/v), acetaldehyde (C₂H₄O, Alfa Aesar, 98%), propionaldehyde (C₃H₆O, Alfa Aesar, 97%), butyraldehyde (C₄H₈O, Alfa Aesar, 98%), valeraldehyde (C₅H₁₀O, TCI Chemicals, India, 95%), hexanal (C₆H₁₂O, TCI Chemicals, India, 95%), crotonaldehyde (C₄H₆O, Alfa Aesar, 98%) and benzaldehyde (C₇H₆O, Alfa Aesar, 99%) were analytically pure and used without further purification. Dulbico's Modified Eagle's Medium with high glucose (DMEM with high glucose, Gibco), PBS (Phosphate-buffered saline, Sigma), paraformaldehyde (Sigma), Rhohamin-Phalloidin (Invitrogen), MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, Sigma], DMSO (dimethyl sulfoxide, Sigma), Trypsin–EDTA solution (HiMedia), Fetal bovine serum (FBS, HiMedia). Milli-Q water was used throughout the experiment.

Cellular uptake of NSCDs was studied by seeding human fibroblast cells at a density of 5 × 10³ on sterile glass cover slips. The cells were then treated with 100 μg mL⁻¹ of NSCDs suspended in DMEM (Dulbecco's Modified Eagle's medium) incomplete medium for different durations (6, 24 and 48 h). After incubation, cover slips were washed with ice-cold PBS and the cells were treated with 3.7% paraformaldehyde for 20 min. Then the actin filaments were stained with Rhohamin-Phalloidin (Invitrogen) dye and then the cells were visualized by confocal laser scanning microscope (CLSM, Olympus FluoView 1000).