Fig. 1. 5-HT stimulated cell morphology alteration in 5-HT6R-EGFP expressed HEK293 cells.
(A) HEK293 cells were transfected with 5-HT6R-EGFP. After 24 h transfection, cells were treated with 1 μM 5-HT for 0, 5, 30 and 60 min. Cells were fixed and immunostained with appropriated mouse anti-GFP antibodies, followed by labeling with anti-mouse FITC (green) or Alexa Fluor555 Phalloidin (red) for F-actin. (B) Cells were pretreated with 20 μM SB258585 for 15 min and then treated with 1 μM 5-HT for 60 min. Right panels of (A) and (B) show the enlarged merged images of the left and middle panels. (C) Cells were pretreated with C3 transferase (C3T, 1 μg/ml), Y-27632 (Y-27, 10 μM), PP2 (20 μM), H89 (20 μM), or PD98059 (PD, 10 μM) for 20 min. After pretreatment, cells were further incubated with 1 μM 5-HT for 60 min. (D) Quantification of morphological changes following each treatment. Cell areas were measured and presented as change of surface area per single cell. Values were normalized to the change of surface area in 5-HT-treated cells. NS: not significant, ***P < 0.001 vs. 5-HT-treated cells.