Fig. 5: LINC00183 directly interacts with ENO1. | Cell Death & Disease

Fig. 5: LINC00183 directly interacts with ENO1.

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

Fig. 5

A Representative results of SDS-PAGE and silver staining assays using pull-down fractions of LINC00183 sense and antisense probes and HT29 cell lysates. B ENO1 was found to be enriched in the fractions that co-eluted with LINC00183 in RNA pull-down assays, according to Western blot analysis. C Western blot analysis confirming that ENO1 was enriched in the fractions co-precipitated with LINC00183 in RIP assays. D Secondary structure of LINC00183, as predicted by RNAfold. E Diagrammatic illustration of the LINC00183 truncated mutant pieces (top panel). Fragment #6 of LINC00183 is necessary for its interaction with ENO1, according to Western blot analysis (bottom panel). F Diagrammatic representation of ENO1 plasmids that are either truncated mutant or FLAG-tagged wild-type (WT). G Western blot analysis revealed that deletion of the 224–404 aa region in ENO1 hindered the ability of LINC00183 to bind ENO1. H RIP assay results showing that deletion of aa 224–404 in the ENO1 protein prevents its binding to LINC00183. I, J IF analysis of ENO1 expression in CRC samples and matched normal colon tissue in tissue microarrays. Scale bar, 200 μm. K and L Correlation between LINC00183 and ENO1 expression. Data are derived from IF analysis of CRC tissue microarrays. M and N Verification of LINC00183 and ENO1 co-localization obtained through RNA-FISH and IF analyses in HT29 and SW480 cells. Scale bar, 20 μm.

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