Fig_1.tif (913.91 kB)
Dominant-negative disruption of SP-R210L.
figure
posted on 2015-05-12, 02:54 authored by Linlin Yang, Marykate Carrillo, Yuchieh M. Wu, Susan L. DiAngelo, Patricia Silveyra, Todd M. Umstead, E. Scott Halstead, Michael L. Davies, Sanmei Hu, Joanna Floros, Francis X. McCormack, Neil D. Christensen, Zissis C. ChroneosRaw264.7 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).
