Journal: eLife
Article Title: LncRNA Snhg3 aggravates hepatic steatosis via PPARγ signaling
doi: 10.7554/eLife.96988
Figure Lengend Snippet: ( A ) Venn diagram of data from RNA pull-down and MS. ( B ) KEGG analysis of genes in specific Snhg3 -binding proteins from RNA pull-down and MS. ( C ) Venn diagram of data from RNA pull-down and MS and bioinformatics predicted by RBPsuite . ( D ) SND1 interacts with different fragments of Snhg3 predicted by bioinformatics using RBPsuite . ( E ) RNA pull-down and western blotting confirms Snhg3 interacting with SND1. ( F ) RIP confirms SND1 interacting with Snhg3 . ( G and H ) Relative protein ( G , up, western blotting; down, quantitative result) and RNA ( H ) levels of Snd1 were measured in the liver. ( I ) Snhg3 enhanced the protein level of SND1 in Hepa1-6 cells (up, western blotting; down, quantitative result). ( J ) Snhg3 promoted the stability of SND1 protein in Hepa1-6 cells (up, western blotting; down, quantitative result). ( K and L ) Snhg3 promoted the ubiquitination of endogenous ( K ) and exogenous ( L ) SND1 protein in Hepa1-6 cells. ( M and N ) Snhg3 increased the K63-linked, not K48-linked and K33-linked, ubiquitination modification of endogenous ( M ) and exogenous ( N ) SND1 protein. ( O ) Snhg3 induced the nuclear localization of SND1 in Hepa1-6 cells (up, western blotting; down, quantitative result). Data are represented as mean ± SEM. *p<0.05and ***p<0.001 by two-way ANOVA ( J ) or Student’s t test (the others). Figure 5—source data 1. Snhg3 -bound proteins were identified in mouse primary hepatocytes by RNA-Pulldown-Mass spectrometry for . Figure 5—source data 2. Snhg3 -bound proteins were predicted by bioinformatic method ( RBPsuite ) for . Figure 5—source data 3. PDF file containing original western blots for , indicating the relevant bands and treatments. Figure 5—source data 4. Original files for western blot analysis displayed in . Figure 5—source data 5. PDF file containing original western blots for , indicating the relevant bands and treatments. Figure 5—source data 6. Original files for western blot analysis displayed in . Figure 5—source data 7. PDF file containing original western blots for , indicating the relevant bands and treatments. Figure 5—source data 8. Original files for western blot analysis displayed in . Figure 5—source data 9. PDF file containing original western blots for , indicating the relevant bands and treatments. Figure 5—source data 10. Original files for western blot analysis displayed in . Figure 5—source data 11. PDF file containing original western blots for , indicating the relevant bands and treatments. Figure 5—source data 12. Original files for western blot analysis displayed in . Figure 5—source data 13. PDF file containing original western blots for , indicating the relevant bands and treatments. Figure 5—source data 14. Original files for western blot analysis displayed in . Figure 5—source data 15. PDF file containing original western blots for , indicating the relevant bands and treatments. Figure 5—source data 16. Original files for western blot analysis displayed in . Figure 5—source data 17. PDF file containing original western blots for , indicating the relevant bands and treatments. Figure 5—source data 18. Original files for western blot analysis displayed in . Figure 5—source data 19. PDF file containing original western blots for , indicating the relevant bands and treatments. Figure 5—source data 20. Original files for western blot analysis displayed in .
Article Snippet: Antibody , anti-Ub (K33) (Rabbit polyclonal) , Abclonal , Cat# A18199; RRID: AB_2861976 , WB (1:1000).
Techniques: Binding Assay, Western Blot, Ubiquitin Proteomics, Modification, Mass Spectrometry
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