Transwell invasion
assays were used to investigate the invasion of B-CPAP cells using
a 24-well transwell chamber (Corning, USA). Briefly, B-CPAP cells
(4 × 104) were seeded into the upper chamber precoated
with Matrigel (BD, USA) and treated with 200 μM ART or LIP@ART.
The lower chamber was filled with 600 μL culture medium. After
24 h, B-CPAP cells on the upper side of the membrane were removed
and cells from seven randomly selected fields on the underside were
imaged and counted under an Axio Vert.A1 fluorescence inverted microscope
(Zeiss, Germany).
A wound healing assay was performed to investigate
the migration of the B-CPAP cells. Briefly, a monolayer of B-CPAP
cells was uniformly seeded in a six-well plate and then wounded by
scratching the surface with a 200-μL pipet tip. After washing
3 times with PBS, the cells were treated with 200 μM ART or
LIP@ART for 24 h. The initial wound and the movement of cells into
the scratched area were imaged at 0 and 24 h under an Axio Vert.A1
fluorescence inverted microscope (Zeiss, Germany). The wound width
was measured in five randomly selected fields, and the migration rate
of the cells was calculated as the wound width at 0 h minus the wound
width at 24 h following normalization by the wound width at 0 h.
assays were used to investigate the invasion of B-CPAP cells using
a 24-well transwell chamber (Corning, USA). Briefly, B-CPAP cells
(4 × 104) were seeded into the upper chamber precoated
with Matrigel (BD, USA) and treated with 200 μM ART or LIP@ART.
The lower chamber was filled with 600 μL culture medium. After
24 h, B-CPAP cells on the upper side of the membrane were removed
and cells from seven randomly selected fields on the underside were
imaged and counted under an Axio Vert.A1 fluorescence inverted microscope
(Zeiss, Germany).
A wound healing assay was performed to investigate
the migration of the B-CPAP cells. Briefly, a monolayer of B-CPAP
cells was uniformly seeded in a six-well plate and then wounded by
scratching the surface with a 200-μL pipet tip. After washing
3 times with PBS, the cells were treated with 200 μM ART or
LIP@ART for 24 h. The initial wound and the movement of cells into
the scratched area were imaged at 0 and 24 h under an Axio Vert.A1
fluorescence inverted microscope (Zeiss, Germany). The wound width
was measured in five randomly selected fields, and the migration rate
of the cells was calculated as the wound width at 0 h minus the wound
width at 24 h following normalization by the wound width at 0 h.