Fig. 5: Effects of exogenous LplA expression on cell phenotypes in IgG-producing CHO cells with or without lipoic acid addition. | Nature Communications

Fig. 5: Effects of exogenous LplA expression on cell phenotypes in IgG-producing CHO cells with or without lipoic acid addition.

From: Boosting energy metabolism and biosynthesis in diverse organisms by a common bacterial salvage lipoylation protein

Fig. 5

Viable cell density (VCD) and viability (VIA) (a), canakinumab titer (b), immunoblotting for lip-DLAT, and lip-DLST from whole cells (c, d), in MTS-GFP and MTS-LplA cells during the cultivation. The solid and dashed lines in (a) represent VCD and VIA, respectively. The relative lipoylation levels in (c, d) are normalized to GAPDH. Data were represented as mean ± SD (n = 3, n biologically independent samples). e Immunoblotting for lip-DLAT and lip-DLST from isolated mitochondria on day 3. The relative lipoylation levels in (e) are normalized to VDAC. Data were represented as mean ± SD (n = 3, n biologically independent samples). Data were representative of at least three independent experiments. f OCR represents mitochondrial respiration on day 3. Data were represented as mean ± SD (n = 13 in MTS-GFP group, n = 12 in MTS-LplA group, n biologically independent samples). g ROS level in MTS-GFP and MTS-LplA cells during the cultivation. Data were represented as mean ± SD (n = 3, n biologically independent samples). h Mitochondrial membrane potential (MMP) in MTS-GFP and MTS-LplA cells during the cultivation. Data were represented as mean ± SD (n = 3, n biologically independent samples). The cell growth (i), canakinumab titer (j), ATP production (k), spare respiratory capacity (l), and immunoblotting for lipoylation of DLAT and DLST on day 3 (m) in MTS-GFP and MTS-LplA cells upon addition of 0, 1, 10 μM lipoic acid. Data were representative of at least three independent experiments. The relative lipoylation levels in (m) are normalized to GAPDH. Data were represented as mean ± SD (n = 4 in (j), n = 7 in (k, l), n = 4 in (m), n biologically independent samples). Lipoic acid and GAPDH antibodies are used as primary antibodies in (c, e, m). Multiple unpaired two-sided t-test was used for the statistical analyses for (b, f, g, h), two-way ANOVA was used for (e), one-way ANOVA was used for (j–l), unpaired two-sided t-test was used for (c, d, m). P adjustments were made for multiple comparisons. DLAT dihydrolipoamide acetyltransferase, DLST dihydrolipoamide succinyltransferase, VDAC voltage-dependent anion channel. Source data are provided as a Source Data file.

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