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. 2020 Jun 16;11:3067. doi: 10.1038/s41467-020-16918-x

Fig. 6. HuR regulates Apob pre-mRNA splicing.

Fig. 6

a RNA pull-down assays were performed using Hepa1–6 cell lysates and in-vitro-transcribed RNAs depicted in Supplementary Fig. 14. p27 5′UTR and CR (coding region) served as positive (P) and negative (N) controls, respectively. A 5-µg aliquot input (Inp.) and binding to TUBB were also assessed. Data are representative from three independent experiments. b Schematic presentation depicting the RNA fragments inserted into the pcDNA3.1 minigene reporters. c The splicing of the pre-RNA expressed from the mini reporters described in b was analyzed in Hepa1–6 cells with silenced HuR, as described (Methods). The pre-RNA (Pre) and mature RNA (RNA) were measured by semi-quantitative PCR analysis. Blank columns, control; Black columns, siHuR. d Hepa1–6 cells were transfected with a HuR siRNA; 48 h later, RNA was prepared and subjected to RT-qPCR analysis to analyze the levels of Apob mRNA (mRNA) and pre-mRNA (Pre). Blank columns, control; Black columns, siHuR. e RNA described in Fig. 2c and Fig. 5c was used to analyze the levels of Apob mRNA pre-mRNA (Blank columns, WT; Black columns, cKO). Data in c (for intron 24) and d are the means ± SD from three independent experiments; Data in e are the means ± SD (Chow: WT, n = 10, cKO, n = 6; HFD: WT, n = 8, cKO, n = 5); significance is analyzed by two-tailed Student’s t-test (c, d) or two-tailed Mann–Whitney U test (e) (*p < 0.05; **p < 0.01). All the error bars are equivalent throughout the figure. Source data are provided as a Source Data file.