Figure 1.

Identification of LncEDCH1

(A and B) Relative LncEDCH1 expression in breast muscles of 7-week-old Xinghua (XH) chicken and White recessive rock (WRR) detected by RNA-seq (A) (n = 1) and real-time qPCR (B) (n = 4). (C) Results of LncEDCH1 5′ RACE and 3′ RACE. (D) Relative LncEDCH1 expression in polyadenylated RNA and total RNA (n = 4). (E and F) Relative LncEDCH1 expression during the proliferation and differentiation of chicken primary myoblasts (CPMs) isolated from WRR (E) (n = 4) and XH chicken (F) (n = 4). (G and H) Tissue expression profiles of LncEDCH1 in 7-week-old WRR (G) (n = 4) and XH chicken (H) (n = 4). The horizontal axis and vertical axis indicate different tissues and their relative expression values, respectively. (I and J) The distribution of LncEDCH1 in the cytoplasm and nuclei of CPMs was determined by qPCR (I) (n = 2) and semi-quantitative PCR (J) (n = 4). GAPDH and U6 served as cytoplasmic and nuclear localization controls, respectively. (K) RNA in situ hybridization of LncEDCH1 in CPM Special FISH probes against LncEDCH1 were modified by Cy3 (red). The nucleus was stained by DAPI (blue). (L) Western blot analysis of the coding ability of LncEDCH1 (n = 1). The potential ORFs of LncEDCH1 were cloned into the pcDNA3.1-3xFLAG-C vector. CPMs transfected with β-actin-3xFLAG were used as a positive control (PC) and untransfected CPMs were used as a negative control (NC). In (L), the numbers shown below the bands are folds of band intensities relative to control. Band intensities were quantified by ImageJ and normalized to GAPDH. Data are expressed as a fold change relative to the control. Results are presented as mean ± SEM. In (A), (B), and (D), the statistical significance of differences between means was assessed using an independent-sample t test (∗∗p < 0.01).