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. 2021 Aug 16;10(8):2098. doi: 10.3390/cells10082098

Figure 2.

Figure 2

Effects of HG and CML on CD147 expression and glycosylation in differentiated adipocytes. Cells were treated with HG (25 mM) or CML (2 µg/mL) with and without tunicamycin (TM, 15 μg/mL) for 24 h. Normal glucose-containing media (control), BSA, and DMSO were used as controls for HG, CML, and TM treatments, respectively. (A) Quantitative assessment of CD147 mRNA levels normalized to the house keeping gene, GAPDH using real-time PCR. (B) Western blot analysis and quantification of the normalized signal intensity of CD147 proteins; GAPDH and β-actin were used as housekeeping proteins. (C) Measurements of soluble CD147 protein in cell culture media using CD147 Quantikine ELISA assay. (D) Representative image and quantification of CD147 protein glycosylation using Pierce™ Glycoprotein Stain. Mouse IgG was used as a control for the immunoprecipitation step. The graphs represent the means ± SD for 3 independent experiments. (* p < 0.05) for comparison with the corresponding control (HG vs. control; CML vs. BSA; TM + HG vs. HG; and TM + CML vs. CML). († p < 0.05) for comparison with the corresponding control in low glycosylated protein (B).