Fig. 3: Effects of exogenous LplA expression on metabolism and mitochondria-related proteins of U2OS cell lines.

a Oxygen consumption rate (OCR) represents for mitochondrial respiration in two engineered cells. Data were represented as mean ± SD (n = 6, n biologically independent samples). b Extracellular acidification rate (ECR) represents for glycolysis in two engineered cells. Data were represented as mean ± SD (n = 6, n biologically independent samples). Specific consumption rates of glucose (c), glutamine (d), pyruvate (e), and lactate (f) in U2OS cells after 10 and 18 h cultivation. Data were represented as mean ± SD (n = 6, n biologically independent samples). g Direction of ¹³C carbon atoms in glycolysis and TCA cycle. Fractional labeling ratio of pyruvate (h), lactate (i), AKG (j), succinate (k), malate (l), and glutamate (m) after 12- and 18-h cultivation of [U-¹³C] glucose addition. Data were represented as mean ± SD (n = 3, n biologically independent samples). n Heatmap of metabolites abundance in glycolysis, TCA cycle and amino acids biosynthesis after 24-h cultivation of [U-¹³C] glucose addition. Data were represented as mean ± SD (n = 8, n biologically independent samples). o Immunoblotting for mitochondrial fusion-related proteins. MFN1, MFN2, OPA1, and GAPDH antibodies are used as primary antibodies. Data were represented as mean ± SD (n = 3, n biologically independent samples). Data were representative of at least three independent experiments. p Immunoblotting for mitobiogenesis-related proteins. NRF1, TFAM, and GAPDH antibodies are used as primary antibodies. Data were represented as mean ± SD (n = 3, n biologically independent samples). Data were representative of at least three independent experiments. q Immunoblotting for mTORC1-related proteins. S6, pS6, S6K1, pS6k1, 4EBP1, p4EBP1, and GAPDH antibodies are used as primary antibodies. Data were represented as mean ± SD (n = 3, n biologically independent samples). Data were representative of at least three independent experiments. Multiple unpaired two-sided t-test was used for the statistical analyses for (a–f, h–m, o–q). P adjustments were made for multiple comparisons. AKG alpha-ketoglutarate, Suc succinate, Mal malate, MFN1 mitofusion1, MFN2 mitofusion2, OPA1 optic atrophy 1, NRF1 nuclear respiratory factor 1, TFAM transcription factor A, S6 ribosomal protein S6, pS6 phosphorylated S6, S6K1 S6 kinase 1, pS6k1 phosphorylated S6K1, 4EBP1 4E-binding protein 1, p4EBP1 phosphorylated 4EBP1. Source data are provided as a Source Data file.