Figure 2.

Over-expression of RAMP2 in HEK293T Cells

(A) HEK293T cells were transfected with a RAMP2 DNA construct that encoded a FLAG epitope tag on the N-terminal end and an OLLAS epitope tag on the C-terminal end. The entire native sequence of RAMP2 except for the initiator Met and predicted cleavable signal peptide was included between the epitope tags. The DNA construct was in a pcDNA3.1+ vector.

(B) Immunoblot of transfected HEK293T cells using antibodies against FLAG (red) and actin (green). RAMP2, cells transfected with RAMP2 DNA construct; MOCK, cells transfected with empty pcDNA3.1 plasmid; control, non-transfected cells.

(C) HEK293T cells transfected with a pIRES2-EGFP vector. Bright field is represented by red channel, and GFP detection is represented by green channel; 48% of the cells were GFP-positive 48 h post transfection. Scale bar, 1 mm.

(D) Borders of cells were traced with the Moore-Neighbor tracing algorithm, based on general localized intensities and their area calculated.

(E) Bar graph of cell area (mean, SEM across all cells in condition) shows no significant difference in cell size across conditions. One-Way-ANOVA p value >0.05.