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. 2024 Aug 5;15:6643. doi: 10.1038/s41467-024-50964-z

Fig. 3. The pinKE mutation impedes Gβγ-mediated effector activation.

Fig. 3

A GIRK1/GIRK2 channel current without (left) and with Gβγ (middle) or Gαβγ (right). B Representative two-electrode voltage clamp (TEVC) recordings in Xenopus laevis oocytes expressing GIRK1/2, Gβ1γ2, and either Gαo (black), GαoK46E (red) or GαoR209C (blue) following treatment with high potassium solution (High K+) and barium (Ba2+), which directly activate and inactivate the channel, respectively. C Maximum voltage differences. Normalized current, normalized to the GIRK 1/2 + Gβγ condition (100%). Data are means ± SD from 3 independent experiments with 6 measurements each; ****p < 0.0001; **p = 0.0083. D GIRK1/GIRK2 channel current with Gαβγ and receptor. E Representative TEVC recordings in Xenopus laevis oocytes expressing dopamine D2 receptor, GIRK1/2, Gβ1γ2, and either Gαo (black), GαoK46E (red), or GαoR209C (blue) following treatment with high potassium solution (High K+) and barium (Ba2+), as well as dopamine, which activates the D2 receptor. F Normalized potassium- and dopamine-induced currents. Data are means ± SD from 3 independent experiments, with 6, 6, and 5 measurements each; ****p < 0.0001. G Normalized currents induced by dopamine divided by currents induced by high potassium (I/Ibasal). Data from (F) are means ± SD; **p = 0.0039. H Representative current recordings from HEK293T cells expressing MOR and the indicated Gαo subunit, showing activation with 1-propanol (PrOH), activation with DAMGO or inhibition with BaCl2. Each solution was delivered for 30 s. I Bar plot shows the ratio of DAMGO-activated to PrOH-activated current for each Gαo subunit. Data are means ± SD from 9 (R209C) or 10 (WT, K46E) independent experiments; ***p = 0.0003; *p = 0.0221. Statistical analysis in C and G performed with one-way ANOVA Tukey’s test. Statistical analysis in F performed by two-way ANOVA with Tukey’s multiple comparison (left) and Sidak pairwise comparison (right) tests. Statistical analysis in I performed with one-way ANOVA Dunett’s test. Source data are provided as a Source Data file.