Fig 1: LncEDCH1 interference destroys Ca2+ homeostasis and inactivates the AMPK pathway to reduce mitochondrial efficiency(A–C) After treatment with 20 µM thapsigargin, change in Fluo-8-AM fluorescence intensity (F - F0/F0) with time (A) (n = 4), maximum fluorescence intensity in (A) and (B) (n = 4), and areas under the curves (AUC) in (A) and (C) (n = 4) with LncEDCH1 interference in vitro. (D–F) Treatment with 10 µM ionomycin evoked change in Fluo-8-AM fluorescence intensity (D) (n = 4), the maximum fluorescence intensity in (D) and (E) (n = 4), and AUC in (D) and (F) (n = 4) after interference of LncEDCH1. (G) Relative basal cytosolic calcium level with LncEDCH1 interference (n = 6). (H–J) Treatment with 10 µM ionomycin evoked change in Fluo-8-AM fluorescence intensity (H) (n = 3), the maximum fluorescence intensity in (H) and (I) (n = 3), and AUC in (H) and (J) (n = 3) induced after co-transfection with the listed nucleic acids. (K–M) Relative mtDNA content (K) (n = 5), mitochondrial membrane potential (L) (n = 6), and intracellular ROS ([ROS]i) (M) (n = 4) with LncEDCH1 interference in vitro. (N–P) Relative enzyme activity of ETC complexes I (N) (n = 3), II (O) (n = 3), and V (P) (n = 3) with LncEDCH1 inhibition in CPMs. (Q and R) Oxygen consumption rate (OCR) (Q) (n = 5), and basal respiration, maximal respiration, and ATP production (R) (n = 5) of myoblasts after interference of LncEDCH1. (S) Protein expression levels of calcineurin and AMPK signaling after the interference of LncEDCH1 in vitro (n = 3). In (S), 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 fold change relative to the control. Results are presented as mean ± SEM. In (B), (C), (E–G), (K–P), and (R), the statistical significance of differences between means was assessed using an independent-sample t test. In (I) and (J), ANOVA followed by Dunnett's test was used (*p < 0.05; **p < 0.01; N.S., no significant difference).