Ophiopogonin D Increases SERCA2a Interaction with Phospholamban by Promoting CYP2J3 Upregulation

Figure 10

The FRET assay of HEK293T cells expressing SERCA2a and PLB. (a) The plasmid profiles of pCMV-N-YFP-SERCA2a and pCMV-N-CFP-PLB. (b) Immunoblot of CFP-PLB, YFP-SERCA2a, and CFP/YFP with anti-PLB monoclonal antibody, anti-SERCA2a antibody, and anti-GFP antibody, respectively. Lanes from the left are (1) pCMV-N-YFP-SERCA2a (predicted band size: 170 kDa), (2) pCMV-N-CFP-PLB (predicted band size: 35 kDa), (3) pCMV-N-CFP-PLB+pCMV-N-YFP-SERCA2a, (4) untransfected HEK293T cell homogenate control, and (5) pCMV-N-YFP + pCMV-N-CFP (predicted band size: 27 kDa). (c) Profiles of YFP-SERCA2a (yellow points) and CFP-PLB (blue points) fluorescence at 10 s after YFP-selective laser spot photobleaching. The data are described in the line chart, and the arrow shows the relationship between YFP-SERCA2a and CFP-PLB fluorescence across the target region. The intensity indicates 18% energy transfer. (d) Profiles of YFP-SERCA2a (yellow points) and CFP-PLB (blue points) fluorescence after OPD treatment. There is no obvious change in the CFP intensity after YFP-selective laser spot photobleaching.

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