TNF protein and mRNA expression levels in empty vector (EV) and ChAT expression vector (ChAT) infected RAW264.7 cells. TNF content in supernatant and in lysates of empty vector and ChAT expression vector infected RAW264.7 cells following incubation for 4 h: (a) untreated cells (PBS instead of LPS). (b) LPS treated cells. Compared with empty vector infected cells, TNF content in the supernatant decreased and in the lysate was increased in ChAT overexpressed RAW264.7cells. The amount of TNF both in supernatant and in lysate was detected by ELISA and shown as OD450. (c) TNF mRNA expression level in ChAT gene transfected RAW264.7 cells was higher than that in empty vector transfected RAW264.7 cells, both in LPS treated and untreated cells. TNF mRNA expression level was determined by real-time PCR. -Actin was used as the control gene. The data shown are representative of three independent experiments and presented as mean SD (). * when compared with EV group.
