Fig. 5. Effect of microtubule-stabilizing drugs on Piezo1 expression and activity.
(A) Piezo1 expression in epidermis of mice treated with paclitaxel or vehicle. (B) Representative cell-attached recordings of stretch-activated currents induced by a stepwise increase of negative pressure in the patch pipette at Vhold = −80 mV. Recordings were made from HEK-P1KO cells expressing GFP (grey –vehicle) and Piezo1-GFP in vehicle (black), and paclitaxel (red) pretreated cells. (C) Current-pressure relationship of stretch-activated currents recorded from HEK-P1KO cells expressing GFP (grey, n=6) and Piezo1-GFP in vehicle- (black, n=36) and paclitaxel treated (red, n=39) cells at a Vhold = −80 mV. (D) The maximal amplitude of stretch-activated Piezo1 current recorded in the vehicle (black, n = 36), and paclitaxel (red, n=39) pretreated HEK-P1KO cells. (E) Representative cell-attached recordings of spontaneous single-channel Piezo1 activity in the vehicle- and paclitaxel-treated HEK-P1KO cells with corresponding all-point amplitude histograms. (F-H) Summary graphs of single-channel Piezo1 amplitude, open probability, and number of channels recorded from vehicle- and paclitaxel-treated HEK-P1KO cells at Vhold= −80 mV. (I) Effect of paclitaxel and ixabepilone treatment on Piezo1 conductance. (J) Current-pressure relationship of stretch-activated currents recorded from Piezo1 expressing HEK-P1KO cells treated with vehicle- (black, n=36) or ixabepilone (1 μM, blue=20). Recordings were made at a Vhold = −80 mV. Statistical analysis for (C and J) was performed using 2way ANOVA with Tukey’s multiple comparisons tests; for (D and H) using the Mann-Whitney test, and for (A, F, G and I) using Student’s t test. Error bars, ± SEM. *P < 0.05, **P < 0.01, and ****P < 0.0001.