The biological
activities of samples isolated from the Korean endemic plant A. spectabilis were evaluated as described in our
previous study with minor modifications.27 (link) HaCaT cells (Korean Cell Line Bank, Seoul, Republic of Korea) were
seeded into 60 mm plates (1 × 106 cells/well) in Dulbecco’s
modified Eagle medium including 5% fetal bovine serum and 1% penicillin
and streptomycin. Samples at a 2.5 μM concentration were treated
in the wells for 2 h and then exposed to UVB at 125 mJ/cm2 using a UV lamp. After UVB irradiation, the cells were harvested,
washed with cold PBS (1×), and lysed in PRO-PREP (iNtRON Biotechnology,
Inc., Sungnam, Republic of Korea). Equal amounts of protein (30 μg)
obtained from the lysed cells were loaded onto 10% SDS gels and then
transferred to PVDF membranes (Millipore, Billerica, MA, USA). The
transferred membranes were incubated with primary antibodies (1:1000)
including GRs (Cell Signaling, MA, USA), 11β-HSD1 (Abcam, Cambridge,
UK), and α-tubulin (Cell Signaling). After 1 day, the attached
primary antibody membranes were incubated with secondary antibodies
(1:2000) for 2 h. All membranes were analyzed using a ChemiDoc XRS+
imaging system (Bio-Rad Laboratories, Inc., Hercules, CA, USA). The
detected band intensity was calculated using Bio-Rad Quantity One
software (ver. 4.3.0, Bio-Rad).
activities of samples isolated from the Korean endemic plant A. spectabilis were evaluated as described in our
previous study with minor modifications.27 (link) HaCaT cells (Korean Cell Line Bank, Seoul, Republic of Korea) were
seeded into 60 mm plates (1 × 106 cells/well) in Dulbecco’s
modified Eagle medium including 5% fetal bovine serum and 1% penicillin
and streptomycin. Samples at a 2.5 μM concentration were treated
in the wells for 2 h and then exposed to UVB at 125 mJ/cm2 using a UV lamp. After UVB irradiation, the cells were harvested,
washed with cold PBS (1×), and lysed in PRO-PREP (iNtRON Biotechnology,
Inc., Sungnam, Republic of Korea). Equal amounts of protein (30 μg)
obtained from the lysed cells were loaded onto 10% SDS gels and then
transferred to PVDF membranes (Millipore, Billerica, MA, USA). The
transferred membranes were incubated with primary antibodies (1:1000)
including GRs (Cell Signaling, MA, USA), 11β-HSD1 (Abcam, Cambridge,
UK), and α-tubulin (Cell Signaling). After 1 day, the attached
primary antibody membranes were incubated with secondary antibodies
(1:2000) for 2 h. All membranes were analyzed using a ChemiDoc XRS+
imaging system (Bio-Rad Laboratories, Inc., Hercules, CA, USA). The
detected band intensity was calculated using Bio-Rad Quantity One
software (ver. 4.3.0, Bio-Rad).
