Skip to main content
. 2015 Jun 8;290(29):18102–18110. doi: 10.1074/jbc.M115.665950

FIGURE 6.

FIGURE 6.

Tid1 interacts with Beclin1. A, transient co-transfection using various combinations of expression plasmids as indicated in the figure was performed in HEK293 cells, followed by GST pulldown assay. B, GST pulldown assay of HEK293 cells co-transfected with FLAG-BECN1 and GST or GST-tagged Tid1L, Tid1S, HSP70, and Tax. C, GST pulldown assay of HEK293 cells co-transfected with PI3KC3-FLAG and GST or GST-tagged protein expression plasmids. D, PI3KC3 knockdown efficiency in HEK293 cells was verified with anti-PI3KC3 blot. E, HEK293 cells stably expressing NS shRNA or PI3KC3 shRNA were transfected with GST or GST-tagged Tid1. F, co-precipitation of Tid1 and PI3KC3 in NS shRNA- or BECN1shRNA-expressing HEK293 cells using transient co-transfection method. G, GST pulldown assay of HEK293 cells co-transfected with FLAG-BECN1 and GST, GST-tagged Tid1L, or its mutants. H, HT1080 cells were transfected with vector, Tid1-FLAG, or its mutants, and the levels of LC3-II were determined with anti-LC3 blot. I, HEK293 cells stably expressing NS shRNA or Tid1 shRNA were co-transfected with Flag-Beclin1 and GST or HSP70-GST. J, co-precipitation of Tid1 with Beclin1 in DMSO or rapamycin (μm/ml, 1 h) HEK293 cells using endogenously expressed proteins. IP, immunoprecipitation.