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Fig. 6. Biochemical and genetic interactions between C. elegans KGB-1
and CSN-5; Drosophila Vasa interacts with KGB-1 and CSN-5.
(A) GST pull-downs using CSN-5-GST and KGB-1-6-His. Lane 1, CSN-5-GST
with eGFP-6-His, negative control (the smaller band is likely to be a CSN-5
breakdown product). eGFP size indicated. Lane 2, CSN-5-GST with KGB-1-6-His.
(B) After csn-5(RNAi) in kgb-1, GLH-1 was assayed by
western blot using 30 worms/lane of: lane 1, uninjected N2 worms; lane 2,
uninjected kgb-1 mutants; lane 3, fertile F1 progeny of
csn-5(RNAi) into kgb-1. All worms assayed were 
3d>L4
stage, 20°C. F1 worms were picked as adults 5-6 days after
moving mothers to fresh plates to eliminate eggs formed before injection
(purging). This experiment was repeated four times, with the decrease in GLH-1
for the fertile kgb-1; csn-5(RNAi) animals averaging
2.8-fold lower than their kgb-1 uninjected siblings (range 1.6-5.3)
(lane 3 versus lane 2). This difference is statistically significant,
P<0.03. ß-tubulin served as a loading control. (C)
Baculoviral co-infections of insect cells treated as in
Fig. 4B,C. Lane 1, Vasa-GST
with eGFP-6-His; lane 2, Vasa-6-His with KGB-1-GST; lane 3, Vasa-6-His with
CSN-5-GST. The His-tagged proteins pulled down are shown in the upper panel,
with the GST proteins shown below. Input proteins for these pull-down assays
are shown in Fig. S3 in the supplementary material.
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