Fig. 1

Effects of Portulaca oleracea ethanol extract (POEE) on receptor activator of NF-κB ligand (RANKL)-induced free cytosolic Ca²⁺ ([Ca²⁺]_i) oscillations and nuclear factor of activated T-cell c1 (NFATc1) amplification. a Isolated bone marrow-derived macrophages (BMMs) were plated on cover glass and cultured for 48 h in the presence of RANKL (50 ng/mL). After incubation, [Ca²⁺]_i was measured using Fura-2 AM fluorescent dye as described in “Materials and Methods.” To confirm the generation of RANKL-induced [Ca²⁺]_i oscillations, cells were initially perfused with regular HEPES buffer, and then acutely treated with 25, 50, and 100 μg/mL of POEE diluted in regular HEPES buffer for the indicated time. Each trace presents the [Ca²⁺]_i response of a single cell. b Isolated BMMs were treated with RANKL (50 ng/mL) for the indicated time with or without POEE (50 μg/mL). Following incubation, whole proteins were collected and used for determining NFATc1 expression. β-actin was used for loading control. NFATc1 expression is shown as the mean of the ratio (NFATc1/β-actin)