Fig. 2.

Presence of NF-SLN proteins in the ER of intact cells and the microsomal fraction from these cells. (A) Subcellular distribution of NF-SLN expression constructs. (B) Transfected cultures were harvested, and broken cell pellet and microsomal fractions were isolated, solubilized, subjected to SDS/PAGE, and probed by using the anti-FLAG antibody. For the constructs SLN 1-22 and SLN 1-23, protein loading was increased to visualize bands. Note that the NF-SLN Y29E/Y31E mutant and those constructs with long C-terminal deletions were virtually absent from the microsomal fraction, but were present in the cell pellet. (C) Ethidium bromide-stained agarose gel of NF-SLN RT-PCR products obtained from RNA isolated from cells transfected with SLN expression constructs, along with a negative control (Neg.).