Fig. 2: Effect of uPSEM⁸¹⁷ on intrinsic electrophysiological properties of the cells expressing PSAM⁴-GlyR.

A Whole-cell patch-clamp recording of a GFP+ cell (green arrow). The Infrared differential interference contrast (IR-DIC) image on the left and fluorescent 470 nm light visualization on the right. Pie chart illustration of the percentage of PSAM⁴-GlyR treated animals ((B), n = 7), and all measured CaMKIIα-PSAM⁴-GlyR-GFP+ cells from those animals ((C), n = 14), undergoing changes in intrinsic properties when uPSEM⁸¹⁷ was washed in. D Test pulse response in the different treatments, green indicating uPSEM⁸¹⁷ application in PSAM⁴-GlyR-GFP+ cells. From left to right, Ri measurement and normalized values in CaMKIIα-PSAM⁴-GlyR-eGFP+ cells (green) and CaMKIIα-eGFP+ cells (Ctrl, orange) before, during, and after uPSEM⁸¹⁷ application. E Differential cell response to 0–100 pA ramps of depolarizing current in different treatments, green indicating uPSEM⁸¹⁷ application in PSAM⁴-GlyR-GFP+ cells. From left to right, number of APs in response to ramps of depolarizing current and normalized values in CaMKIIα-PSAM⁴-GlyR-eGFP+ cells (green) and CaMKIIα-eGFP+ cells (Ctrl, orange) before, during, and after uPSEM⁸¹⁷ application. F Differential cell response to the lowest depolarizing current pulses for AP induction abbreviated as Imin (in this case 80 pA) in the different treatments, green indicating uPSEM⁸¹⁷ application in PSAM⁴-GlyR-GFP+ cells. From left to right, number of APs in response to Imin current pulses and normalized values in CaMKIIα-PSAM⁴-GlyR-eGFP+ cells (green) and CaMKIIα-eGFP+ cells (Ctrl, orange) before, during, and after uPSEM⁸¹⁷ application. G Differential cell response to 500 pA depolarizing current pulses in the different treatments, green indicating uPSEM⁸¹⁷ application in PSAM⁴-GlyR-GFP+ cells. From left to right, number of APs in response to 500 pA pulses and normalized values in CaMKIIα-PSAM⁴-GlyR-eGFP+ cells (green) and CaMKIIα-eGFP+ cells (Ctrl, orange) before, during, and after uPSEM⁸¹⁷ application. Ri, input resistance; Imin, minimum current pulse step to trigger an AP. Animals injected with AAV8-CaMKIIα-PSAM⁴-GlyR-IRES-eGFP, n = 7 (total cells n = 14); control/GFP-only animals injected with AAV8- CaMKIIα-eGFP, n = 3 (total cells n = 7). Cells used in the analysis as responders n = 12. Median ± interquartile range. Wilcoxon paired test for comparison of the PSEM effect to the Baseline. **, p < 0.01.