Cellular impedance measurements
confirm the function of GPR35 in
HepG2 cells. HepG2 cells cultured in xCELLigence microtitre plate
wells were able to measure alterations of electrical impedance over
time. (A) Once basal conditions were established, vehicle (red), iodoxamide
(green) (upper and middle panels), CID-2745687 (pink) (upper panel),
or ML-145 (pink) (middle panel) was added either alone or in combination
with lodoxamide (blue) (upper and middle panels) and alterations in
the signal measured over time. Lower panel: Difference in the “cell
index” measured at 5 min treatment with the effect of lodoxamide
presented as 100%. (a) p < 0.05 versus lodoxamide.
(B) Various inhibitors of signaling pathways were added (FR = FR900359,
1 × 10–7 M) (upper panel), PTX = Pertussis
toxin, 100 ng/mL (middle panel), and Y27632, 1 × 10–5 M (lower panel). FR and Y27632 were added 30 min before lodoxamide
and PTX 24 h before. Right-hand panels: Difference in the “cell
index” measured at 5 min treatment with the effect of lodoxamide
presented as 100%. a: p < 0.05 versus lodoxamide.