Fig. 6: LINC00183 stabilizes ENO1 by inhibiting its degradation through the ubiquitin-proteasome system. | Cell Death & Disease

Fig. 6: LINC00183 stabilizes ENO1 by inhibiting its degradation through the ubiquitin-proteasome system.

From: Platelet-derived exosomal LINC00183 facilitate colorectal cancer malignant progression driven by histone lactylation through stabilizing ENO1

Fig. 6

A, B HT29 and SW480 cells’ relative mRNA and protein ENO1 expression after LINC00183 overexpression or knockdown were examined using RT-qPCR and western blot techniques. C, D RT-qPCR and western blot analyses of ENO1 expression in HT29 and SW480 cells co-cultured with CRC/PLT-Exos. E, F ENO1 expression in CRC cells treated with or without MG132 (10 μmol/L for 10 hours) was examined using Western blot. G–J Western blot analysis ENO1 expression after LINC00183 overexpression or silencing in CHX-treated CRC cells. K–N Time course of changes in relative ENO1 protein levels. O, P Co-IP and western blot analyses examining ENO1 ubiquitination in HT29 cells co-transfected with His-tagged ubiquitin, FLAG-tagged ENO1, and either si-LINC00183, LINC00183, or the corresponding controls. TCL: total cell lysate. Q Co-IP and western blot analyses of ENO1 ubiquitination levels in HT29 cells transfected with plasmids expressing FLAG-tagged WT or mutant (K224-227A, K228A, K262A, and K281A) ENO1 protein along with His-tagged ubiquitin plasmids and LINC00183-targeted siRNA or control siRNA. R Plasmids expressing FLAG-tagged ENO1 or FLAG-tagged mutant ENO1 (K262A) were used to transfect HT29 and SW480 cells. The LINC00183 probe was then used in a lncRNA pull-down assay, and a particular anti-FLAG antibody was used for western blot analysis.

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