Effect of DHA on TLR4 expression, cytotoxicity, and cell
migration in vitro. (a) Representative confocal images
of TLR4 expression
patterns in murine 4T1 cells after being treated with 10 μM
DHA for 24 h followed by 4 μg mL–1 of LPS
stimulation. Scale bar = 10 μm. (b) Fluorescence intensity per
cell of TLR4 on 4T1 cells is analyzed by ImageJ. (c) At a constant
DOX concentration of 1.72 μM, dose–response histograms
of 4T1 cells without (left) or with (right) 4 μg mL–1 of LPS prestimulation for 72 h against different molar ratios of
DHA and DOX for 72 h. (d) Dose–response curves of 4T1 cells
untreated (left) or treated (right) with 4 μg mL–1 of LPS prestimulation for 72 h against DOX, DHA, and DOX + DHA for
72 h. (e) Apoptosis rate (left) and late apoptosis rate (right) of
4T1 cells untreated or treated with 4 μg mL–1 of LPS prestimulation for 72 h against DOX, and DOX + DHA with an
equivalent concentration of 5.00 μg mL–1 of
DOX and 3.12 μg mL–1 of DHA for 4 h. (f, g)
Representative images of 4T1 (left) and RAW264.7 (right) prestimulation
with 4 μg mL–1 of LPS for 72 h or not were
taken by phase contrast microscope before and after treatment with
1.00 μg mL–1 of DOX or 0.57 μg mL–1 of DHA. Closure rates were obtained by ImageJ software
(below). Data represent means ± SD (n = 3). P values were determined by one-way ANOVA with post
hoc Tukey test and represented using *P <
0.05, **P < 0.01, ***P < 0.001,
and ****P < 0.0001. n.s., not significant.