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. 2023 May 15;13(8):3365–3381. doi: 10.1016/j.apsb.2023.05.010

Figure 4.

Figure 4

VCR potentiated PIEZO2 MA currents in rat DRG neurons and PIEZO2 transfected HEK 293T cells. (A) Sample traces showed the PIEZO2 MA currents evoked by a 7-μm membrane displacement (above) at the initial time (black) and 30 min (red) after establishing whole-cell mode patch recording with (right, VCR) or without (left, Cont) intracellular administration of VCR (5 μmol/L). The traces also included continual membrane tests by voltage steps of 5 mV at the interval of 100 ms (light color). (B) Summary data of the potentiation of PIEZO2 MA currents recorded with (red circle, VCR) or without (black square, Cont) intracellular application of VCR (5 μmol/L) over time. n = 17 and 10 in Cont and VCR group, respectively. (C) Representative traces of PIEZO2 MA currents elicited by a set of membrane displacements from 1 to 7 μm (above the current traces) in control group (Cont, above the dish line) and VCR intracellular treatment group at different concentrations (VCR, below the dish line). (D) Statistics for the potentiation of PIEZO2 MA currents treated intracellularly by different concentrations of VCR. The currents were evoked at 30 min and VCR was used at 0.2 μmol/L (black diamond, n = 4), 1 μmol/L (black inverted triangle, n = 4), 5 μmol/L (black circle, n = 5) and 25 μmol/L (black triangle, n = 4), respectively. n = 7 in Cont group. (E) Concentration–effect curve of VCR-induced potentiation of PIEZO2 MA currents in rat DRG neurons. The potentiation fold was evaluated from the effects induced by different concentrations of VCR at the 7-μm stimulation displacement. The EC50 is shown above. (F) Representative traces of PIEZO2 MA currents elicited by a 7-μm poking displacement (above) at the initial time (black trace) and 20 min (red trace) from 5 μmol/L VCR intracellularly treated DRG neurons who were incubated with scrambled shRNA (left) or PIEZO2 shRNA (right) lentivirus. (G) Summary data of PIEZO2 MA currents evoked with different membrane displacements at 20 min in scrambled shRNA group (black square, n = 9) or PIEZO2 shRNA group (red circle, n = 10) of 5 μmol/L VCR intracellularly treated DRG neurons. (H) Representative traces of PIEZO2 currents elicited by a set of membrane displacements from 1 to 10 μm (above the current traces) at the initial time (black traces) and 15–20 min (red traces) in control group (above the dish line) and in 5 μmol/L VCR intracellular treatment group (below the dish line, VCR) in PIEZO2 transfected HEK 293T cells. In VCR group, the grey traces represented PIEZO2 currents were extracellularly treated with 10 μmol/L D-GsMTx4 at 15–20 min. (I) Summary data of PIEZO2 currents evoked with different membrane displacements at 15–20 min in control group (black square, n = 5), VCR-treatment group (red circle, n = 4) and VCR + D-GsMTx4 group (grey triangle, n = 5) of PIEZO2 transfected HEK 293T cells. Data are shown in mean ± SEM; #/$/∗P < 0.05, ##/$$/∗∗P < 0.01, ###/$$$/∗∗∗P < 0.001.