Research Paper Volume 14, Issue 15 pp 6028—6046

Ascorbic acid induces salivary gland function through TET2/acetylcholine receptor signaling in aging SAMP1/Klotho (-/-) mice


Figure 6. TET2 upregulated muscarinic receptor and salivary gland functional protein expression in PSGC kl +/+ cells. PSGC kl +/+ cells were transfected with the TET2 expression plasmid (pcDNA3-TET2-Flag) for 48 hrs. After transfection, qRT–PCR and Western blotting were performed to evaluate the mRNA and protein levels. (AD) Expression of M1 and M3 muscarinic receptors (M1 and M3AchR) in PSGC kl +/+ cells overexpressing TET2. (EH) Effect of TET2 on α-amylase, aqua5, and ZO-1 expression. Data are presented as the mean ± SD. *p < 0.05, **p < 0.01. (I) Immunocytochemistry staining of ZO-1 and Aqua5 in TET2-overexpressing PSGC kl +/+ cells. PSGC kl +/+ cells were transfected as above for 48 h. After transfection, anti-mouse Flag antibody and Alexa Flour 488-labeled sheep anti-mouse secondary antibody were used to detect TET2 proteins. Aqua5 and ZO-1 were detected by primary anti-rabbit Aqua5 and ZO-1 antibodies and Alexa Flour 488-labeled sheep anti-rabbit secondary antibodies (I). Bar: 100 μm. Images were observed with a confocal microscope.