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Fig. 6 | Journal of Experimental & Clinical Cancer Research

Fig. 6

From: Tripartite motif-containing 14 (TRIM14) promotes epithelial-mesenchymal transition via ZEB2 in glioblastoma cells

Fig. 6

Depletion of TRIM14 increased ZEB2 polyubiquitination and proteasomal degradation. a qTR-PCR analysis of TRIM14 and ZEB2 mRNA level in LN229 and U251 cells after TRIM14 knowdown by shTRIM14. Depletion of TRIM14 does not alter ZEB2 mRNA. ***p < 0.001, n = 3 experiments. b Western blot analysis of TRIM14, ZEB2 and β-actin in LN229 and U251 cells transduced with the indicated shRNA in the absence or presence of 10μM MG132. c LN229 and U251 cells transfected with shTRIM14 were treated with cycloheximide (100μg/ml, pretreated for 15 min and for varying durations), and collected at the indicated times for western blot. Quantification of ZEB2 expression relative to β-actin is shown. Results are shown as mean ± standard deviation. n = 3 independent experiments. ***, P < 0.001, two-way ANOVA test. d LN229 and U251 cells transfected with HA-UB and the indicated shRNA were treated with MG132 for 8 h before harvest. ZEB2 was immunoprecipitated with anti-ZEB2 antibody and immunoblotted with anti-HA antibody. e LN229 and U251 cells were transduced with the indicated shRNA for 48 h, treated with the proteasome inhibitor MG132 for 6 h, and then subjected to analysis of ZEB2 ubiquitination and IB analyses. f HS683 and U87 cells were transduced with the Myc-TRIM14, Flag-FBXO45 or vector control for 48 h, treated with the proteasome inhibitor MG132 for 6 h, and then subjected to analysis of ZEB2 ubiquitination and IB analyses

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