Fig. 4: LINC00842 directly interacts with PGC-1α in PADC cells.

a, b Subcellular localization analysis by PCR (a) or RNA FISH (b) shows LINC00842 is mainly located in the nuclei. U6 and GAPDH were served as nuclear and cytoplasmic markers and shown are mean ± SD from 3 independent experiments (a). Scale bar, 30 μm (b). c Western blot analysis of proteomic screening suggested 8 potential LINC00842-binding proteins obtained from RNA pull-down assays with LINC00842 or its antisense. d Association of PGC-1α with LINC00842 determined by RNA immunoprecipitation (RIP) assays. Data represent relative enrichment (mean ± SD) to input from 3 independent experiments. IgG was used as the negative control. e Results of chromatin isolation by RNA purification (ChIRP) assays using LINC00842-odd, -even or LacZ (negative control) antisense probe sets. Upper panels are RNA retrieval rate (mean ± SD from 3 assays) and lower panels are immunoblot of PGC-1α and GAPDH (negative control) in ChIRP and input. f Immunofluorescence assays show co-localization of LINC00842 (red) and PGC-1α (green) predominately in the nuclei. Scale bar, 30 μm. g Truncation mapping of LINC00842 PGC-1α binding domain. Top panel: diagrams of LINC00842 full-length and truncated fragments. Middle: RNA sizes of in vitro transcribed LINC00842 full-length and truncated fragments. Bottom: immunoblot analysis of PGC-1α pulled down by different LINC00842 fragments. h Schematic diagram of Flag-tagged PGC-1α and its truncated forms used in LINC00842 pull-down assays. i Immunoblot analysis of Flag-tagged wild-type (WT) PGC-1α and its truncated forms retrieved by in vitro transcribed biotinylated LINC00842.