Extended Data Fig. 1: NEAA inhibits Neat1 expression in hepatocytes.
From: Proline exacerbates hepatic gluconeogenesis via paraspeckle-dependent mRNA retention
a, Differential lncRNAs expression analysis in NEAA-treated (NEAA) vs. control (Ctrl) hepatocytes. b, Expression levels of the top ten significantly altered lncRNAs in NEAA-treated hepatocytes, as determined by BioGPS analysis (www.biogs.org). c, Schematic diagram illustrating the two splice variants of Neat1, with the positions of primers used for qPCR or probes for FISH indicated. d, Neat1_1/2 and Neat1_2 levels in NEAA-treated hepatocytes relative to controls, n = 4 independent experiments. e, Representative fluorescence images of cells stained for Neat1 (green) RNA in control and NEAA-treated hepatocytes. Nuclei are stained with DAPI (blue). f, Quantification Neat1 dots in hepatocytes, n = 40 cells per group. Box plots are as described in Fig. 1. g, The dose-dependent effect of proline on Neat1 level in hepatocytes from male and female mice, n = 3 independent experiments. h, Nono mRNA levels in control siRNA (siScr) and Nono knockdown (siNono) hepatocytes, n = 3 independent experiments. i, Control or Nono knockdown hepatocytes were treated with or without proline for 24 h and then exposed to actinomycin D (ActD) at indicated time points to assess the half-lives of Neat1, n = 3 independent experiments. Data are presented as means ± SD. Statistical significance was determined using a two-tailed Student’s t test in (a), (b), (d), (f) and (h); one-way ANOVA in (g); two-way ANOVA in (i).
