Fig. 5: Tob interacts with TNF-receptor complex I-associated proteins. | Cancer Gene Therapy

Fig. 5: Tob interacts with TNF-receptor complex I-associated proteins.

From: Tob negatively regulates NF-κB activation in breast cancer through its association with the TNF receptor complex

Fig. 5

a MCF-7 parental or Tob KO were treated with 10 ng/mL TNF-α at indicated times. Cell lysates were subjected to immunoprecipitation with anti-Tob antibody. Whole cell lysates (WCL) were also prepared from the same cell lysates before performing immunoprecipitation. b HEK 293 T cells were transiently transfected with C-terminal Myc-tagged Tob with NEMO, N-terminal Flag-tagged TRAF2, N-terminal Flag-tagged RIPK1 or N-terminal Flag-tagged CYLD. After 24 h, cells were harvested and were subjected to immunoprecipitation with anti-Myc tag antibody. WCLs were also prepared from the same cell lysates before performing immunoprecipitation. c Tob+/+ or Tob-/- MEFs were stimulated with Flag-tagged TNF-α (2 μg/mL). Cells were harvested at the indicated times and were subjected to immunoprecipitation with anti-Flag tag antibody-conjugated beads. WCLs were also prepared with the same cell lysates before performing immunoprecipitation. For the 0 min samples, 2 μg/mL of Flag-tagged TNF-α were added in post-lysis. d HEK 293 T cells were transiently transfected with C-terminal Myc-tagged Tob plasmids with N-terminal Flag-tagged RIPK1 parental or K377R mutant plasmids. After 24 h, cells were harvested and were subjected to immunoprecipitation as described in (b).

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