Fig. 5

Gαq T96S mutant acts in a dominant negative manner to promote tumor growth of NKTCL. a Western blot shows that tetracycline-inducible cell lines (KHYG1 and YT) express equal amounts of wild-type and mutant Gαq simultaneously. b Viability of YT and KHYG1 cells with inducible expression of vector, wild-type, T96S or wild-type plus T96S. Cell viability is presented as the absorbance value at OD 450 nm. Data are representative of three independent experiments. c Apoptosis was induced after starving KHYG1 cells of IL-2 for 12 h and was assessed by Annexin V staining assay. Data are representative of three independent experiments. d Representative images of xenograft tumors (left) and tumor weights (right) for YT cells with inducible expression of vector, wild-type, T96S or wild-type plus T96S (n = 5 in each group). e Schematic diagram of the approach used to identify Gαq-interacting partners using immunoprecipitation in combination with mass spectrometry. Significance analysis of INTeractome (SAINT) expression was utilized to calculate the probability of protein–protein interactions from background, nonspecific interactions. The right panel shows representative binding proteins identified by mass spectrometry. f Confirmation of the interaction of wild-type Gαq or Gαq T96S mutant with Gβ1 by immunoprecipitation assay. The Gαq T96S mutant bound Gβ1 more tightly than wild-type Gαq in YT cells. All data are expressed as the mean ± s.e.m.; NS, not significant. *P < 0.05, **P < 0.01, ***P < 0.001, unpaired two-tailed Student’s t-test. Source data are provided as a Source Data file