Fig 2: lncRNA ZFP36L2-AS interacts with ACACA and PC to induce ACACA dephosphorylation and inhibit PC protein stabilization.A, B lncRNA ZFP36L2-AS interacts with ACACA and PC protein were determined by biotin-labeled RNA pulldown (A) and RIP (B). C The interaction of full-length and truncated ZFP36L2-AS with ACACA and PC protein was determined by RNA pulldown. D–G The protein expression levels of ACACA and phosphorylated ACACA (D, F), and relative ACACA activity (E and G) after ZFP36L2-AS overexpression or knockdown in vitro and in vivo. H–K The protein expression level of PC (H, J), and relative PC activity (I, K) after ZFP36L2-AS overexpression or knockdown in vitro and in vivo. (L) The protein expression level of PC in ZFP36L2-AS overexpressed myoblast was analyzed after incubated with the protein synthesis inhibitor cycloheximide (CHX; 25 µg/ml). (M) The protein expression level of PC in ZFP36L2-AS knockdowned myoblast was analyzed after incubated with the proteasome inhibitor (MG-132; 5 µmol/L) for 12 h. N, O Relative acetyl-CoA content (N), and relative PC activity (O) with ACACA interference in vitro. P Relative acetyl-CoA content with ZFP36L2-AS overexpression or knockdown in vivo. Q, R Relative acetyl-CoA content (Q), and relative PC activity (R) induced by the listed nucleic acids. In panels (D, F, H, J, L, and M), the numbers shown below the bands were folds of band intensities relative to control. Band intensities were quantified by ImageJ and normalized to ß-Tubulin. Data are expressed as a fold-change relative to the control. Results are presented as mean ± SEM. In panels (E, G, I, K and N–P), statistical significance of differences between means was assessed using independent sample t-test. In panels (Q, R), ANOVA followed by Dunnett’s test was used. (*P < 0.05; **P < 0.01; N.S., no significant difference).