Dominant-negative disruption of SP-R210L.
Raw264.7 cells were stably transfected with empty pTriexNeo2 control or vector containing the 300 (210L(DN1)) and 350 (210L(DN2)) bp cDNA of SP-R210 carboxy-terminal isoforms (6). A) Detergent extracts were analyzed by Western blotting using affinity purified polyclonal antibodies recognizing both SP-R210L and SP-R210S. Lanes were loaded with 5 μg of protein. B) Total RNA from indicated cell lines was reverse transcribed and quantitated by qRT-PCR using TaqMan assays and primers encompassing the SP-R210L-specific exons 1 and 2 (red bars) and internal primers encompassing exon 17 and 18 common to both SP-R210 isoforms (black bars) and 18S rRNA as internal control. (n = 4 ***p<0.001).