Figure 1.
Sequences of mutations detected and family trees of the patients with each base substitution.
(A) Japanese family of MRX214 with Del 312F in GPR37. (B) Caucasian family of AU0654 with mutation R558Q in GPR37. (C) Caucasian family with T589M in GPR37. Closed boxes and circles indicate the individual with ASD, + indicate the individual with mutations.
Table 1.
Results of the analysis of GPR37.
Figure 2.
The location of the GPR37 mutations.
Arrow indicates the position of Del312F, R558Q and M589T.TM, transmembrane domain.
Figure 3.
The localization of wild-type and mutated GPR37 in C2C5 cells.
(A) Co-localization of wild-type GPR37-myc with endoplasmic reticulum (ER)-marker (Bip/GRP78). Scale bar: 10 µm. (B) C2C5 cells were transfected with wild-type GPR37-myc, GPR37(R558Q) -myc or GPR37(Del312F)-myc for 28 h after transfection. Cells were fixed, and immunostained with anti-myc (GPR37, red) and Hoechst (blue). Scale bar: 20 µm.
Figure 4.
Appearance of apoptotic cells and cell having intracellular accumulation of GPR37(R558Q) and GPR37(Del312F) in time-dependent manner.
(A) Apoptotic cells expressing wild-type GPR37-myc, GPR37(R558Q)-myc or GPR37(Del312F)-myc. Green, GPR37 wild-type or mutants. Blue, Hoechst. Right pictures, high magnification of left pictures. Arrowheads, apoptotic morphology. Scale bars: 40 µm. (B) The percentage of the apoptotic cells in the cells expressing, GPR37(R558Q)-myc or GPR37(Del312F)-myc. Values are mean ± standard error (SEM). All experiments were performed three times. A significant difference of the percentage of cells exhibiting apoptotic morphology was detected in the cells expressing wild-type GPR 37 and mutated GPR37. (Student's t-test; p<0.05). Control, cells expressing vacant vector (GFPC1-vector).
Table 2.
Primer sequences of GPR37.