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Fig. 6. Arabidopsis protein interactions analyzed by yeast two-hybrid
and coimmunoprecipitation assays. (A) Interactions between SUF3,
AtSWC6 and AtSWC2 (SWC2). (B) Interactions between SUF3, AtSWC6, AtSWC2
and HTA8, HTA9 and HTA11. (C) The same interaction analysis as B but
with baits and preys changed. (D) Interaction between regions of PIE1
and SUF3, AtSWC6, AtSWC2, HTA8, HTA9 and HTA11. PIE1-N, PIE1-M and PIE1-C
indicate PIE1 regions comprising amino acids 1-520, 520-1220 and 1220-2055,
respectively. (E) The same interaction analysis as D but with baits and
preys changed. (F) Protein gel blots showing interaction between AtSWC6
and SUF3. IP indicates immunoprecipitation of protein extracts from tobacco
leaves transiently co-expressing epitope-tagged proteins, as listed above the
blot, with anti-GFP (
-GFP) or anti-Flag (-Flag) antibodies, as
indicated below the blot. About 10% of the total sample used in each IP was
loaded as an input control. Mock indicates IP with anti-Flag antibody. The
immunoprecipitates and protein extracts were separated by 9% SDS-PAGE,
transferred to polyvinylidene difluoride membranes, and probed with anti-myc
antibody. (G) Protein gel blots showing interaction between AtSWC6,
AtSWC2 and HTA11. Tobacco leaves co-expressing myc:SUF4 and AtSWC6:GFP were
used as a negative control of IP. The protein blots were probed with anti-GFP
antibody. (H) Protein gel blots showing interaction between PIE1 and
SUF3 and AtSWC6. Mock indicates IP with no antibody. The protein blots were
probed with anti-Flag (left) or anti-myc (right) antibodies.
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