Fig. 1: Targeted recruitment of YFP-tagged NEDD4-2 HECT domain to reconstituted CaV2.2 inhibits channel surface density and ionic currents. | Nature Communications

Fig. 1: Targeted recruitment of YFP-tagged NEDD4-2 HECT domain to reconstituted CaV2.2 inhibits channel surface density and ionic currents.

From: Ion channel inhibition by targeted recruitment of NEDD4-2 with divalent nanobodies

Fig. 1: Targeted recruitment of YFP-tagged NEDD4-2 HECT domain to reconstituted CaV2.2 inhibits channel surface density and ionic currents.The alternative text for this image may have been generated using AI.

a Schematic of BBS-α1B + β + α2δ-1 + YFP + nb.F3 + nb.YFP. Cell surface α1B channels are selectively detected using Alexa Fluor 647-conjugated bungarotoxin (BTX-647) attaching to an extracellular bungarotoxin binding site (BBS) engineered into α1B. Created in BioRender. Colecraft, H. (https://BioRender.com/crkk1ct). b Exemplar flow cytometry contour and cumulative density frequency (CDF) plots. c Exemplar whole-cell currents for cells expressing α1B + β + α2δ-1 + YFP + nb.F3 + nb.YFP. d Population I-V curve for cells expressing α1B + β + α2δ-1 + YFP + nb.F3 + nb.YFP. Data points are means ± SEM; n = 8 for each point from three separate transfections. e Schematic; created in BioRender. Colecraft, H. (https://BioRender.com/crkk1ct). f Flow cytometry data for cells expressing BBS-α1B + β + α2δ-1 + YFP + DiVa nb[F3-YFP]. g, h Exemplar currents and population I-V curve for cells expressing α1B + β + α2δ-1 + YFP + DiVa nb[F3-YFP], n = 9 from three independent transfections (red traces, symbols and curves). Black trace in (h) is I-V curve reproduced from (d) to facilitate visual comparison. i Schematic; created in BioRender. Colecraft, H. (https://BioRender.com/crkk1ct). j Flow cytometry data for cells expressing BBS-α1B + β + α2δ-1 + YFP-HECT4-2 + nb.F3 + nb.YFP. (k, l) Exemplar currents and population I-V curve for cells expressing α1B + β + α2δ-1 + YFP-HECT4-2 + nb.F3 + nb.YFP. n = 8 from three independent transfections (blue traces, symbols and curve). Black trace in (l) is I-V curve reproduced from (d) to facilitate visual comparison. m Schematic; created in BioRender. Colecraft, H. (https://BioRender.com/crkk1ct). n Flow cytometry data for cells expressing BBS-α1B + β + α2δ-1 + YFP-HECT4-2 + DiVa nb[F3-YFP]. o, p Exemplar currents and population I-V curves for cells expressing α1B + β + α2δ-1 + YFP-HECT4-2 + DiVa nb[F3-YFP], n = 9 from three independent transfections (magenta traces, symbols and curve). Black trace in (p) is I-V curve reproduced from (d) to facilitate visual comparison. *p = 0.0398 by one-way ANOVA and Tukey’s multiple comparisons test.

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