Fig. 3

Upregulation of HLA-DMB expression on HPMCs after exposure to PYS or to HPG at day 3 and day 7. The mean of fluorescence intensity (MFI) of HLA-DMB staining on HPMCs after repeating exposure to PYS or to HPG at day 3 and day 7 compared to untreated control at day 0 (as a baseline) was measured by using flow cytometric analysis. The ratio of MFI on treated cells to its baseline was presented as fold change of HLA-DMB expression. a Data were presented as a typical histogram of HLA-DMB stain (solid line) in each group compared to the baseline (MFI: 126, dotted line), showing the fold change of HLA-DMB expression. b Data were presented as mean ± SD of four separate experiments (n = 4). p = 0.0036 (PYS vs. HPG, two-way ANOVA). *p = 0.0011 (PYS: day 3 vs. day 7, t-test). **p = 0.0032 (HPG: day 3 vs. day 7, t-test). (C) The total cellular levels of HLA-DMB staining on HPMCs after repeating exposure to PYS or to HPG at day 3 and day 7 compared to untreated control at day 0 were determined by Western blot analysis. Equal amount of protein (100 to 150 μg) extracted from whole cell pellets was fractioned by 10% of SDS-PAGE, and HLA-DMB protein bands were identified based on specifically binding of anti-HLA-DMB antibody, and their molecular size (26–28 kDa) (upper panel). The protein content in each sample was confirmed by re-probing the blot with anti-GAPDH antibody (middle panel) and was measured by densitometry. Imaging data are a representative of three separate experiments. The ratio of HLA-DMB band to GAPDH band from the same sample on the same blot was presented (bottom panel)