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. 2017 Jul 24;8:864. doi: 10.3389/fimmu.2017.00864

Figure 7.

Figure 7

Expression of LAT1 and 4F2hc is induced upon T cell stimulation even under limiting arginine concentrations. Primary human CD3+ T lymphocytes were isolated from blood of healthy donors by negative selection. T cells were stimulated in medium containing 1,000, 20, or 0 µM arginine for 6, 24, or 48 h or left unstimulated (0 h). LAT1 (A) and 4F2hc (B) mRNA expression levels were quantified by quantitative real-time reverse transcription polymerase chain reaction. B2M (β2-microglobulin) was used for relative measurements. Data are expressed as percentage of the respective LAT1/B2M or 4F2hc/B2M ratio obtained in the same experiment for cells stimulated for 48 h in 1,000 µM arginine [mean ± SD, (A) n = 4–5, (B) n = 3–4]. (C,D,E) LAT1, 4F2hc, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) protein expressions were determined by Western blot. (C) Representative blot, (D) quantification of LAT1, and (E) 4F2hc protein. Data are expressed as percentage of the respective LAT1/GAPDH or 4F2hc/GAPDH ratio obtained in the same experiment for cells stimulated for 48 h in 1,000 µM arginine (mean ± SD, n = 3). For statistical analysis each time point of stimulated T cells was compared to unstimulated control cells (0) (***p < 0.001, **p < 0.01,*p < 0.05), two-way analysis of variance with Tukey’s posttest.