Fig 1: In vitro LPE P-18:0 treatment elevates intracellular cAMP levels and activates PKA signalling via inhibition of PDE activity.a, b Immunoblot analysis of C3H10T1/2 adipocytes treated with vehicle control (VC), LPC P-18:0 (LPC-P, 10 µM) or LPE P-18:0 (LPE-P, 10 µM) for 24 h or isoproterenol (ISO, 10 µM) for 4 h. n = 4. c, d Immunoblot analysis of C3H10T1/2 adipocytes treated with VC, ISO (10 µM), LPC-P (10 µM), LPE-P (10 µM), ISO (10 µM) + H89 (PKA inhibitor, 50 µM), LPC-P (10 µM) + H89 (50 µM), or LPE-P (10 µM)+H89 (50 µM). n = 3. (p-HSL/HSL VC vs. ISO: p < 0.000001; VC vs. LPC-P: p = 0.000128; VC vs. LPE-P: p = 0.000009; ISO vs. ISO + H89: p < 0.000001; LPC-P vs. LPC-P + H89: p = 0.000056; LPE-P vs. LPE-P + H89: p = 0.000006)(p-CREB/CREB VC vs. ISO: p = 0.000035; VC vs. LPC-P: p = 0.000018; VC vs. LPE-P: p = 0.000019; ISO vs. ISO + H89: p = 0.000172; LPC-P vs. LPC-P + H89: p = 0.000143; LPE-P vs. LPE-P + H89: p = 0.000109). e Intracellular cAMP levels in C3H10T1/2 adipocytes treated with VC, LPC-P (10 µM), or LPE-P (10 µM) for 24 h or ISO (10 µM) for 4 h. n = 3. f Levels of 5' AMP released by PDE3B incubated with VC, IBMX (40 µM), or LPE-P (10 µM) for 90 min. n = 3. g, h Immunoblot analysis of C3H10T1/2 adipocytes treated with VC, LPC-P (10 µM) or LPE-P (10 µM) for 48 h or isoproterenol (ISO, 10 µM) for 24 h. n = 4. i, j Oxygen consumption rate measurement in C3H10T1/2 adipocytes treated with VC, LPC-P (10 µM) or LPE-P (10 µM) for 24 h. n = 5. Each point represents a biological replicate. Data are presented as mean ± SEM. Statistical significance was determined by the unpaired, two-tailed t-test in b, d, e, f, h and j. * represents statistical analysis between C3H10T1/2 adipocytes treated with VC and C3H10T1/2 adipocytes treated with either ISO, LPC-P, or LPE-P. # represents statistical analysis between C3H10T1/2 adipocytes treated with either ISO, LPC-P, or LPE-P and C3H10T1/2 adipocytes treated with either ISO + H89, LPC-P + H89, or LPE-P + H89. Source data are provided as a Source Data file.
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