Preparation and Treatment of HEK Cells

U73122 (S8011, Selleck Chemicals, Houston, TX, USA) is dissolved in dimethyl sulfoxide (DMSO, D8371, Beijing, China), with the final concentration of 9 mM, and stored at −20 °C. For subsequent experiments, combined with cell viability results (see Supplementary data: Supplementary Fig s9a) and previous research results^{49 (link),54 (link)}, we selected 9 μM U73122 treatment of HEK for 15 min.
MK-2206 (2HCl) (S1078, Houston, TX, USA) is dissolved in dimethyl sulfoxide (DMSO, D8371, Beijing, China), with the final concentration of 12 mM, and stored at −80 °C. For subsequent experiments, combined with cell viability results (see Supplementary data: Supplementary Fig s9a) and previous research results^{55 (link)}, we selected 12 μM MK-2206 treatment of HEK for 24 h.
Pertussis toxin (PTX, P7208, Sigma-Aldrich St., Louis, USA) is dissolved in ddH₂O, with the final concentration of 200 μg/ml, and stored at 4 °C. For subsequent experiments, combined with cell viability results (see Supplementary data: Supplementary Fig s9a) and previous research results^{48 (link)}, we selected 200 ng PTX treatment of HEK for 4 h.
Fluo-3 AM (F1241, Invitrogen, Thermo Fisher Scientific, USA) is dissolved in dimethyl sulfoxide (DMSO, D8371, Beijing, China), with the final concentration of 3 mM and stored at the light-free conditions at −20 °C.