Ceramide induces Txnip mRNA and protein expression. (A) 10I cells were treated with 40 μM C2-ceramide for 2 hours. Cells were collected and total RNA was purified followed by cDNA synthesis. Total RNA (left) and cDNA (middle) were confirmed by agarose gel electrophoresis. After microarray analysis, the results of hybridization in the cDNA chip (right) revealed the up-regulated (red; arrowhead) and down-regulated (green) genes. (B) The effects of C2-ceramide on the mRNA expression of Txnip via a time- and dose-dependent manner were determined by RT-PCR. The expression of β-actin was used as an internal control. (C) The protein expression of Txnip and thioredoxin after various doses of C2-ceramide treatment was detected in 10I and Jurkat T cells using Western blot analysis. The α-tubulin expression was used as an internal control. The relative ratios of Txnip–α-tubulin and thioredoxin–α-tubulin are shown.
