Fig. 2

EGR1 is critical for human LL-37-enhanced ASC migration and proliferation. a EGR1 mRNA expression was determined by real-time PCR. EGR1 mRNA was detected after treatment with 10 and 20 μg/mL LL-37 for 0–6 hr. b ASCs were incubated with 20 μg/mL LL-37 for 0–6 hr. Cell lysates were collected for western blot analysis. The EGR1 protein level was increased by LL-37 treatment in a time-dependent manner. c ASCs were treated with LL-37 and transfected with siRNA targeting EGR1 or a negative control sequence. After stabilization, cells were collected and lysed by the addition of 200 μL of cell lysis buffer. EGR1-targeting siRNA transfection was quantified by performing an EGR1-specific ELISA of cell lysates. d Cells that migrated after EGR1-targeting siRNA transfection were imaged by microscopy and counted using Scanscope. Results are expressed as the mean ± SD of three independent experiments. *, P < 0.01 vs. control; ^§, P < 0.01 vs. LL-37-treated cells transfected with negative control siRNA. e The effects on human ASC proliferation were similar to those on migration. A representative experiment from three independent experiments is shown. Bars represent the mean ± SD. *, P < 0.01 vs. control; ^§, P < 0.01 vs. LL-37-treated cells transfected with negative control siRNA. EGR1 early growth response 1, ASC adipose-derived stromal/stem cells, siRNA small interfering RNA, ELISA enzyme-linked immunosorbent assay