Fig 2: Expression of neuroendocrine markers in P-STS cells. (A) Light microscopy picture of P-STS cells adhering to the culture dish, (B) Immunofluorescence staining with anti-CgA (Cy3-labeled secondary antibody) and anti-synaptophysin (Alexa 488-labeled secondary antibody) after 6 months of continuous cultivation, (C) 5-HT in the supernatant after 1 h of incubation of adherent P-STS cells (continuously cultivated for 10 months) in serum-free medium containing 5% BSA at room temperature and in the cell lysate as determined by ELISA. *p < 0.05 as calculated by the unpaired two-tailed Student’s t-test assuming equal variances. (D) Expression of neuroendocrine marker genes in P-STS cells assessed by real-time PCR analysis. Real-time PCR analysis was performed using the ?Ct method for relative quantification. Expression mRNA levels of the target gene were normalized to the average of the HKGs. Shown are genes encoding neuroendocrine markers such as CHGA (encoding chromogranin A), SYP (encoding synaptophysin), TPH1 (encoding tryptophan hydroxylase 1), and TAC1 (encoding the tachykinin peptide hormone family, substance P and neurokinin A, as well as the related peptides). Results are shown as mean values in one experiment ± SD and are representative of two independent experiments.

Fig 3: Comparison of serum CgA levels in subgroups of subjects divided according to UACR and healthy subjects. (& P < 0.05, && P < 0.001 vs. healthy controls; ** P < 0.001 vs. normoalbuminuria group; # P < 0.05 vs. microalbuminuria group)

Fig 4: Roc curve for validity of serum CgA in prediction of early DN