Figure 3.
Figure 3. TRPM8 stimulation inhibits cell adhesion. (Ai) Representative photographs showing the “rounding-up” effect induced by treatment with icilin for 10 min (right). (Aii) Quantification of cell adhesion area in HMECs treated with icilin (white circles) or not (black circles) and plated on FN. The experiment was performed using the XCellIgence-Roche apparatus. The gray bar indicates values relative to the stage represented in Ai. (Bi) Simulation of the rounding-up effect in a virtual cell expressing basal levels of 0.2 µM TRPM8, treated with icilin. (Bii) Evolution in time of the relative area of a simulated cell depleted of TRPM8 (CNTRL) or expressing basal levels of TRPM8 and treated with icilin (TRPM8+icilin) or not (TRPM8). The gray bar indicates values relative to the stage represented in Bi. (C) Quantification of adherent HMECs treated or not with 10 µM icilin on plates coated with 0, 0.25, 0.5, and 1.0 µg/ml FN. *, P < 0.05. (D) Quantification of haptotaxis of HMECs treated or not with 10 µM icilin on 1 µg/ml FN or control substrate. *, P < 0.05. (E) Quantification of adhesion to FN of HMECs silenced for TRPM8 (HMEC/siTRPM8) or not (HMEC), in the presence or absence of icilin. (F) Quantification of adhesion to FN of HUVECs silenced for TRPM8 (HUVEC/siTRPM8) or not (HUVEC), in the presence or absence of icilin. (G) Quantification of adhesion to FN of BTECs overexpressing TRPM8 (BTEC/TRPM8) or not (BTEC), in the presence or absence of icilin. *, P < 0.05. All data in C–G represent adhesion normalized to the control untreated condition. Data are expressed as mean ± SEM.

TRPM8 stimulation inhibits cell adhesion. (Ai) Representative photographs showing the “rounding-up” effect induced by treatment with icilin for 10 min (right). (Aii) Quantification of cell adhesion area in HMECs treated with icilin (white circles) or not (black circles) and plated on FN. The experiment was performed using the XCellIgence-Roche apparatus. The gray bar indicates values relative to the stage represented in Ai. (Bi) Simulation of the rounding-up effect in a virtual cell expressing basal levels of 0.2 µM TRPM8, treated with icilin. (Bii) Evolution in time of the relative area of a simulated cell depleted of TRPM8 (CNTRL) or expressing basal levels of TRPM8 and treated with icilin (TRPM8+icilin) or not (TRPM8). The gray bar indicates values relative to the stage represented in Bi. (C) Quantification of adherent HMECs treated or not with 10 µM icilin on plates coated with 0, 0.25, 0.5, and 1.0 µg/ml FN. *, P < 0.05. (D) Quantification of haptotaxis of HMECs treated or not with 10 µM icilin on 1 µg/ml FN or control substrate. *, P < 0.05. (E) Quantification of adhesion to FN of HMECs silenced for TRPM8 (HMEC/siTRPM8) or not (HMEC), in the presence or absence of icilin. (F) Quantification of adhesion to FN of HUVECs silenced for TRPM8 (HUVEC/siTRPM8) or not (HUVEC), in the presence or absence of icilin. (G) Quantification of adhesion to FN of BTECs overexpressing TRPM8 (BTEC/TRPM8) or not (BTEC), in the presence or absence of icilin. *, P < 0.05. All data in C–G represent adhesion normalized to the control untreated condition. Data are expressed as mean ± SEM.

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