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Fig. 6. Subcellular localization of transiently transfected NF-SLN and NF-PLN cDNAs in HEK-293 cells. (A) Immunofluorescence analysis of cells cotransfected with NF-SLN and SERCA2a in a 1:1 ratio and examined 48 h using the anti-FLAG and anti-SERCA antibodies. Note the colocalization between NF-SLN and SERCA. (B) HEK-293 cells were transfected with single plasmids encoding NF-SLN, NF-PLN, or the SLN mutant Y29E/Y31E. Note the characteristic ER staining pattern with NF-SLN that is largely absent in the SLN mutant. (Bar, 40 mm.) (C) Extracts were collected from a whole-cell lysate, microsomes, or a broken cell pellet, subjected to SDS/PAGE, and probed with the anti-FLAG antibody.

Fig. 7. Immunofluorescence analysis of cells cotransfected with NF-SLN and SERCA2a. Cells were cotransfected with SLN expression constructs together with SERCA2a in a 1:1 ratio and examined after 48 h using the anti-FLAG and anti-SERCA antibodies. Shown are representative examples of overlay staining of FLAG and SERCA2a in cells cotransfected with SERCA2a and one of NF-SLN 1-25, 1-26, 1-27, 1-28, 1-29, or NF-SLN Y29E/Y31E. Note the precise colocalization of SERCA2a and the SLN constructs. (Bar, 40 m m.)