Fig 1.
Chemical structure of BSG.
Table 1.
RT-PCR primer sequences.
Fig 2.
Effect of BSG on levels of ceramides in the SC of RHEK models.
A: HPTLC chromatogram of lipids in the SC; B: Levels of total ceramides; C: Levels of ceramide species. RHEK models were treated for 5 days in culture without or with BSG (1, 3 and 10 μg/mL). The extraction of lipids from the SC of the RHEK model and HPTLC analysis were performed as described in Materials and methods section. Data are expressed as means ± SD (n = 6). *p<0.05.
Fig 3.
Effect of BSG on mRNA expression levels of enzymes involved in ceramide synthesis in RHEK models.
A: SPT2, n = 3; B: CerS3, n = 4; C: GCS, n = 4; D: GBA, n = 4; E: SMS2, n = 4; and F: ASM, n = 4. RHEK models were cultured in the absence or presence of BSG at the indicated concentrations for the indicated number of days. Real time RT-PCR analysis was performed as described in the Materials and methods section. Data are expressed as means ± SD. *p<0.05, **p<0.01.
Fig 4.
Effect of BSG on mRNA expression levels of enzymes involved in ceramide synthesis in HaCaT keratinocytes.
A: SPT2, n = 4; B: CerS3, n = 4; C: GCS, n = 4; and D: SMS2, n = 4. HaCaT cells were cultured in the absence or presence of BSG at the indicated concentrations for the indicated number of hours. Real time RT-PCR analysis was performed as described in the Materials and methods section. Data are expressed as means ± SD from 4 independent experiments. *p<0.05, **p<0.01.
Fig 5.
Effect of BSG on protein expression levels of enzymes involved in ceramide synthesis in HaCaT keratinocytes.
A: SPT2, n = 5; B: CerS3, n = 5; C: GCS, n = 5; and D: SMS2, n = 5. HaCaT cells were cultured in the absence or presence of BSG at the indicated concentrations for 48 hr. Western blotting analysis was performed as described in the Materials and methods section. Representative immunoblots from 5 experiments are shown. Data are expressed as means ± SD from 5 independent experiments. *p<0.05, **p<0.01.
Fig 6.
The mechanism of SC ceramide increasing effect of BSG.
Increased levels of expression by BSG are indicated by red arrows.