Figure 2.

TMEM120A does not inhibit PIEZO1 and TREK1 currents. (A) HEK293 cells were transfected with Piezo1 in IRES-GFP vector, with or without Tmem120a. Mechanically activated currents were evoked by increasing indentations with a blunt glass probe in whole-cell patch-clamp experiments. Current amplitudes are plotted (mean ± SEM) for Piezo1 expressing cells (black) and for cells expressing Piezo1 and Tmem120a (red). (B) Scatter plots and mean ± SEM for current amplitudes at 5.2 μm indentation. Statistical significance was calculated with the Mann-Whitney test. (C) Representative current traces. (D) Piezo1 deficient N2A cells were transfected with GFP-Piezo1 with or without Tmem120a-tdTomato, and with tdTomato-Tmem120a alone or tdTomato alone. Mechanically activated currents were evoked by applying increasing negative pressures through the patch pipette in cell-attached patch-clamp experiments. Measurements were performed at −80 mV holding potential. Current amplitudes are plotted (mean ± SEM) for cells expressing Piezo1 (black), Piezo1 and Tmem120a (red), Tmem120a alone (orange), and tdTomato alone (green). (E) Scatter plots and mean ± SEM for current amplitudes at −55 mmHg. Statistical significance was calculated with the Mann-Whitney test. (F) Representative current traces. (G) Piezo1 deficient Neuro2A cells were transfected with TREK1 with, or without Tmem120a-tdTomato, and with Tmem120a-tdTomato alone or tdTomato alone. Mechanically activated currents were evoked by applying increasing negative pressures through the patch pipette in cell-attached patch-clamp experiments. Measurements were performed at 0 mV holding potential. Current amplitudes are plotted (mean ± SEM) for cells expressing TREK1 (black), TREK1 and Tmem120a (red), Tmem120a alone (orange), and tdTomato alone (green). (H) Scatter plots and mean ± SEM for current amplitudes at −55 mmHg. Statistical significance was calculated with the Mann-Whitney test. (I) Representative current traces.