Figure 8. Notch signaling assay in asthmatic naïve CD4⁺T cells.

A, Asthmatic CD4⁺T cells were transfected with Lfng plasmid and RBPJ-κ luciferase reporter plasmid pGa981-6. Then the luciferase activities were analyzed and normalized to Renilla luciferase activity in control group, asthmatic group, asthmatic CD4⁺T cells treated with control vector pEGFP-N1 and asthmatic CD4⁺T cells treated with Lfng, Mfng or Rfng group. We found that Notch signaling was activated in asthmatic group and overexpression of Lfng led to a reduction in Notch signaling activity. But there was no significant differences in Mfng or Rfng group. The results are shown as mean±SEM from three samples. *p<0.05; **p<0.01. The results are from one representative experiment of three independent experiments. B, Realtime PCR was performed to detect the mRNA levels of Hes-1. The mRNA levels of control CD4⁺T cells group were taken as 1. Results are from three independent experiments. The data for each group are expressed as means±SEM. *p<0.05, significant differences between asthmatic group with control group, or asthmatic group with asthmatic CD4⁺T cells transfected with Lfng, Mfng or Rfng group. The Hes-1 mRNA decreased in Lfng group comparing with the asthmatic counterparts, but there was no significant differences in Mfng or Rfng group. C, Hes-1 protein levels in each group. Lane 1, control CD4⁺T cells; Lane 2, asthmatic CD4⁺T cells; Lane 3, asthmatic CD4⁺T cells transfected with pEGFP-N1 plasmid; Lane 4, asthmatic CD4⁺T cells transfected with Lfng plasmid. Representative of one of three similar experiments. The Hes-1 protein decreased in Lfng treated group.