Fig. 3

Effects of RS on rat liver CerSs and mitochondrial ceramides. A-D RT-PCR results demonstrated that RS induced an increase in CerS2, CerS5, and CerS6 in the liver (n = 6–8). Values were normalised to the geometric mean of two housekeeping genes (TUBB2a and Ubc) and presented as fold change relative to control values, as evaluated by the ΔΔCT method. E–F Western blot results revealed that RS increased the protein expression of CerS5 and CerS6 in the liver (n = 3–4). G Western blotting was employed to detect the contents of VDAC (a mitochondrial outer membrane marker), calnexin (an ER marker), and GM130 (a Golgi marker) in both isolated mitochondria and cytoplasm to assess the purity and integrity of the isolated mitochondria. H RS induced an increase in C16:0 ceramide content in isolated mitochondria (n = 5). Ceramide species content was measured by LC–MS/MS. Data were analysed by the Mann–Whitney test or unpaired t-test. ^* P < 0.05, ^** P < 0.01, compared to the control group