Journal: International Journal of Nanomedicine
Article Title: LincRNA-p21: A Double-Edged Sword in Ethanol-Induced Liver Damage and Its Nanoparticle Solution
doi: 10.2147/IJN.S577455
Figure Lengend Snippet: ALKBH5/lincRNA-p21/FGFR2 promotes autophagy in response to ethanol-induced liver injury. ( A ) HepG2 cells were treated with/without 10 μM SAHA, 10 μM NaB, or 5 μM RGFP966 for 24 h, the expression of lincRNA-p21 was evaluated by RT-PCR. n=3. ( B ) The potential m 6 A sites in lincRNA-p21 predicted using SRAMP. ( C ) m 6 A RNA methylation levels of HepG2 cells with/without alcoholic hepatocyte injury ( P >0.05). n=6. ( D ) When HepG2 cells were exposed to ethanol, the expressions of lincRNA-p21, ALKBH5 and METTL3 were evaluated. The expressions of lincRNA-p21 and ALKBH5 were all down-regulated when HepG2 cells were exposed to 100 mM ethanol ( P <0.01, P <0.05). ( E ) The location of lincRNA-p21 in HepG2 cells demonstrated by the lincRNA-p21 FISH Probe. Scale bars: 50 μm. ( F ) The expressions of lincRNA-p21 was upregulated by ALKBH5 overexpression. n=3. ( G ) LincRNA-p21 was significantly enriched in ALKBH5-bound RNA. n=3. ( H )The methylation level of the 1237 nt site in lincRNA-p21 was evaluated using MeRIP-PCR. n=3. ( I ) Schematic diagram of ALKBH5 mediating m 6 A demethylation and upregulation of lincRNA-p21. The red downward arrow indicates decreased expression. ( J ) The mRNA and protein expression of FGFR2 was evaluated by RT-PCR and Western blot. FGFR2 protein was up-regulated by the overexpression of lincRNA-p21 (FGFR2/β-actin: 0.54 vs 0.20, P =0.001). ( K ) Fluorescence images of the autophagosomes. Scale bars: 10 μm. ( L ) Cell viability was evaluated using a CCK8 assay. n=3. Data represent mean±s.e.m. Significance was determined by two-tailed Student’s t -test. * P <0.05, ** P <0.01, *** P <0.001.
Article Snippet: The HDAC inhibitors Vorinostat (SAHA)(#HY-10221, MCE, USA), sodium butyrate (NaB)(#S1999, Selleck, USA), and HDAC3 inhibitor RGFP966 (#HY-13909, MCE, USA) were used to study the mechanism of lincRNA-p21 regulation.
Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Methylation, Over Expression, Western Blot, Fluorescence, CCK-8 Assay, Two Tailed Test
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