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Fig. 6. BNIPXL targets proto-Lbc via two distinct sites and suppresses Lbc-induced cellular transformation. (A) Schematic diagram of proto-Lbc and its truncation mutants. 
-HEL, -helical region; PP, proline-rich motif; CT, extreme C-terminus; 
C, oncogenic Lbc. Lysates expressing Flag-proto-Lbc, mutants or vector control with HA-BNIPXL (B) full-length, (C) CBCH, (D) WBCH or (E) NBCH were subjected to immunoprecipitation and bound proteins detected with anti-HA (first panels) and anti-Flag (second panels) to show the amounts of precipitated proteins. Expression of BNIPXL (third panels) and proto-Lbc (fourth panels) was verified. Asterisks indicate position of the expected band. Arrow indicates the position of the nonspecific bands arising from anti-HA antibody. (F) Schematic diagram indicating a requirement for the DH-PH domains and the proline-rich motif for BNIPXL-Lbc interactions. (G) NIH3T3 was transfected with DsRed-onco-Lbc and GFP-BNIPXL fragments in primary focus formation. Foci were stained with crystal violet after 21 days and the number of foci was counted. Transforming activity of onco-Lbc was set at 100%. Data are means ± s.d. (n=3). Differences between values not sharing the same letters (a, b, c, d) are statistically significant at P<0.05 by analysis of variance and the Student-Newman-Keuls multiple range test (Statgraphics).
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