Fig. 3

Hypoxia and inflammation presence modify HMEC-1 migration. (a) Top panel, representative image depicting the 3D microfluidics device design. A collagen matrix (2.5 mg/mL) was added to the central channel and HMEC-1 cells were added to the side channel. Middle panel, 3D-rendering of the microfluidic device. Bottom panel, higher-power magnification image showing cell migration. A gradient of chemoattractant for cell migration was established by adding 50% (v/v) fetal bovine serum (FBS) to the right media channel. Chips were treated with 24 µM FKBPL siRNA or 1 mM DMOG or 50% MCM, ± 100 nM AD-01. After 72 h, chips were probed with antibodies and subjected to immunofluorescence imaging to determine the expression of CD31 and DAPI. As control for siRNA treatment, non-targeted siRNA was used. Untreated cells were used as controls for the DMOG and MCM treatments. (b) Representative images of CD31 and DAPI stained HMEC-1 following FKBPL siRNA treatment ± AD-01. (c) FKBPL siRNA, (d) DMOG and (e) MCM treatment significantly increased HMEC-1 migration whereas AD-01 suppressed only siRNA FKBPL- and DMOG-induced migration. (f) Representative images of CD31 and DAPI stained HMEC-1 cells following DMOG/MCM treatment ± AD-01. Scale bars represent 100 μm. The data were plotted as the mean ± SEM; every data point represents one microfluidic device containing three experimental units, n = 3. The data passed the Shapiro-Wilk normality test and were analyzed by one-way ANOVA with Tukey’s post-hoc test; *p < 0.05, ***p < 0.001 (compared to control); ^#p<0.05, ^##p<0.01 (compared to stimuli conditions)