Fig. 5: PTEN-K260 deacetylation by SIRT7 contributes to NEDD4L-mediated ubiquitination. | Nature Communications

Fig. 5: PTEN-K260 deacetylation by SIRT7 contributes to NEDD4L-mediated ubiquitination.

From: SIRT7 facilitates endometrial cancer progression by regulating PTEN stability in an estrogen-dependent manner

Fig. 5

A Immunoblot manifesting PTEN protein level and the quantitative analysis for three repeats (n = 3). SIRT7 was overexpressed in HEC-1B cells, and the cells were treated with DMSO or MG132 (10 μM, 8 h) or CHQ (20 μM, 16 h) before being collected. The error bars indicate the s.e.m. values. The P value was calculated with two tailed unpaired t test. B Ubiquitination level of PTEN with or without SIRT7 overexpression in HEK293FT cells. C Ubiquitination level of PTEN in sgSIRT7 (pool) or sgScramble HEK293FT cells. D Ubiquitination level of PTEN-WT/ K260Q/ K260R. For B–D, the HA-Ub and FLAG-EV, FLAG-PTEN-WT/K260Q/K260R were overexpressed in HEK293FT cells as indicated, and the ubiquitination level was detected after immunoprecipitation. E Mass spectrum results suggesting the E3 ligase interacting with PTEN and ranked by number of unique peptides. F PTEN protein level shown by immunoblots after NEDD4L knockdown. NEDD4L was deleted in HEC-1B cells by short hairpin RNAs. G Ubiquitination level of FLAG-PTEN after NEDD4L knockdown. HA-Ub and FLAG-PTEN were over expressed, and the ubiquitination level was detected after immunoprecipitation. H Ubiquitination level of FLAG-PTEN after NEDD4L knockdown and SIRT7-Myc overexpression. HA-Ub and FLAG-PTEN were over expressed, and the ubiquitin level was detected after immunoprecipitation. I Co-IP manifesting the interaction between endogenous NEDD4L and PTEN after Myc-SIRT7 overexpression in HEK293FT cells. J Co-IP manifesting the interaction between endogenous NEDD4L and PTEN-WT/K260Q after Myc-SIRT7 overexpression in HEK293FT cells.

Back to article page