Fig. 4: Molecular mechanism of CCNE1 stabilization of ANLN protein expression. | Cell Death Discovery

Fig. 4: Molecular mechanism of CCNE1 stabilization of ANLN protein expression.

From: CCNE1 stabilizes ANLN by counteracting FZR1-mediated the ubiquitination modification to promotes triple negative breast cancer cell stemness and progression

Fig. 4

A Pearson correlation analysis from TCGA demonstrated a significant positive correlation between CCNE1 and ANLN expression levels. B Triple immunofluorescence staining of CCNE1 (red), ANLN (green), and the cell nucleus (DAPI, blue) demonstrated that CCNE1 and ANLN primarily colocalize within the nuclei of TNBC cells. C Endogenous co-immunoprecipitation assay from TNBCcells revealed protein interactions between CCNE1 and ANLN. D Western blot experiment conducted on TNBC cells demonstrated that CCNE1 has the ability to enhance the expression of ANLN protein. E The stability of ANLN protein was tested after treatment of CCNE1-knockdown BT-549 and MDA-MB-231 cells with the 40 μM protein synthesis inhibitor cycloheximide (CHX). F Upon treatment with the proteasome inhibitor 10 μM MG132, ANLN protein stability was tested in CCNE1-knockdown BT-549 and MDA-MB-231 cells. G Ubiquitination assay of ANLN in CCNE1-knockdown BT-549 and MDA-MB-231 cells treated for 4 h with 10 μM MG132. H The E3 ligases that can regulate ANLN substrates were searched based on UbiBrowser2 database. I, J Ubiquitination assay of ANLN in (H) FZR1-knockdown or (I) CCNE1/FZR1 knock down coexisting BT-549 and MDA-MB-231 cells treated for 4 h with 10 μM MG132.

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