Validation of the selected genes. (A) Representative Western blot detection of RUNX1, CTSL1, BATF, BASP1, KDSR, NOTCH1, VDR, and ATP1B1 at 0, 4, 12, 24, 48, and 72 hour time points after culturing of cord blood CD4⁺ cells in Th17-polarizing medium or Th0 control condition. Histone H2B was analyzed to confirm equal loading. (B) Flow cytometric detection of CD52, IL2RB, CXCR5, LMNA, and ITM2A after culturing for 48 and 72 hours. Analysis of the stained cells was done with an LSRII flow cytometer (BD Biosciences). (C) RT-PCR detection of MIAT, BTBD11, and COL6A3 at 24-, 48-, and 72-hour time points. EF1α was used as an endogenous normalization control. Fold changes were calculated by comparing the normalized expression values to the corresponding expression in Th0 control samples at the 24-hour time point. SEM, statistical significance were determined with Student t test, and the average fold change is shown in the figure.