Fig. 6: Chisel-1 selectively eliminates recombinant CaV2.2 channels reconstituted with CaVβ1. | Nature Communications

Fig. 6: Chisel-1 selectively eliminates recombinant CaV2.2 channels reconstituted with CaVβ1.

From: Selective posttranslational inhibition of CaVβ1-associated voltage-dependent calcium channels with a functionalized nanobody

Fig. 6

a Schematic showing modular structures of Nedd4L and Chisel-1. b Schematic of α1C + β1b ± Chisel-1. c Left, β1b immunoblot in untransfected cells (UT), or cells transiently transfected with α1C + β1b and CFP, nb.E8 or Chisel-1. Right, Bar charts showing impact of Chisel-1 on normalized β1b expression (n = 5 independent experiments each for CFP, nb.E8, and Chisel-1 groups). d Left, β1b pulldown followed by β1b immunoblot. Middle, β1b pulldown followed by ubiquitin immunoblot. Right, Bar chart showing Chisel-1 enhances ubiquitination of β1b (n = 5 independent experiments each for CFP, nb.E8, and Chisel-1 groups). e Left, α1C immunoblot. Right, impact of Chisel-1 on α1C normalized expression (n = 6 independent experiments each for CFP, nb.E8, and Chisel-1 groups). f Left, α1C pulldown followed by α1C immunoblot. Middle, α1C pulldown followed by ubiquitin immunoblot. Right, Bar chart showing Chisel-1 enhances ubiquitination of α1C (n = 6 independent experiments each for CFP, nb.E8, and Chisel-1 groups). g Exemplar family of whole-cell currents in HEK293 cells expressing α1B+β1b+α2δ−1 in the absence (left) or presence (right) of Chisel-1. h Population current density vs voltage (J-V) relationship for cells expressing α1B+β1b+α2δ−1 in the absence (black circles; n = 20 over 5 independent transfections) or presence (red circles; n = 10 over three independent transfections) of Chisel-1. i J-V relationship for α1B+β2b+α2δ−1 ± Chisel-1, same format as (h). (For CFP, n = 8 over 3 independent transfections, black symbols; for Chisel-1, n = 6 over 3 independent transfections, red symbols). j J-V relationship for α1B+β3+α2δ−1 ± Chisel-1, same format as (h) (For CFP, n = 17 over 3 independent transfections, black symbols; for Chisel-1, n = 8 over 3 independent transfections, red symbols). k J-V relationship for α1B+β4+α2δ−1 ± Chisel-1, same format as (h) (for CFP, n = 7 over 3 independent transfections, black symbols; for Chisel-1, n = 10 over 3 independent transfections, red symbols). Data are means ± SEM. Source data are provided as a Source Data file.

Back to article page