Fig. 6: DIAPH2-AS1 upregulates NTN1 expression by interacting with NSUN2. | Cell Death & Disease

Fig. 6: DIAPH2-AS1 upregulates NTN1 expression by interacting with NSUN2.

From: Long noncoding RNA DIAPH2-AS1 promotes neural invasion of gastric cancer via stabilizing NSUN2 to enhance the m5C modification of NTN1

Fig. 6

a SDS-PAGE and silver staining using fractions pull downed by DIAPH2-AS1 and its antisense were performed using the lysis of HGC-27 cells. b The secondary mass spectrometry of NSUN2. c Western blot analysis confirmed that NSUN2 was enriched in the fractions pulled down by DIAPH2-AS1. d RIP assay verified that the antibody against NSUN2 could retrieve DIAPH2-AS1. e Relative mRNA and protein levels of NTN1 were detected by qRT-PCR and western blot in indicated HGC-27 cells. f The secondary structure of DIAPH2-AS1 was predicted by catRAPID (http://service.tartaglialab.com/). g The schematic diagram of the truncated mutant fragments of DIAPH2-AS1 (upper panel). Western blot analysis showed that the truncated fragment #5 of DIAPH2-AS1 was indispensable for the interaction between DIAPH2-AS1 and NSUN2 (bottom panel). h The diagrammatic sketch of FLAG-tagged WT or truncated mutant plasmids of NSUN2. i Western blot analysis indicated that the DIAPH2-AS1 failed to enrich NSUN2 after deleting the region of 462–614aa. j RIP assay showed that the fragment of NSUN2 without 462–614aa lost the binding capacity with DIAPH2-AS1. Data and error bars were shown as mean ± SD of triplicate independent replicate experiments and all data were analyzed by Student’s t test (*P < 0.05; **P < 0.01; ***P < 0.001).

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