Di-(2-ethylhexyl)-phthalate interferes with T-follicular helper cell differentiation and cytokine secretion through signaling lymphocytic activation molecule family member-1

Figures & data

Figure 1. Effects of DEHP exposure on OVA-sensitized weanling mice spleen T_fh cell surface receptor SLAMF1 expression. (A) Expression percentage of CXCR5⁺SLAMF1⁺ T_fh cells gated on CD4⁺ T-cell populations (%) (flow cytometry). (B) SLAMF1 mRNA expression (normalized to Gapdh; real-time qRT-PCR). Data shown are means ± SD, n = 8 mice/group. **p < 0.01, ***p < 0.001 vs. corn oil + OVA control group.

Figure 2. Effects of DEHP exposure on OVA-sensitized weanling mice spleen T_fh cell adaptor protein SAP expression. (A) SAP protein expression (ratio to GAPDH; Western blot). (B) SAP mRNA expression (normalized to Gapdh; real-time qRT-PCR). Data shown are means ± SD, n = 8 mice/group. ***p < 0.001 vs. corn oil + OVA control group.

Figure 3. Influence of Slamf1 siRNA transfection (with DEHP exposure) on T_fh cell adaptor protein SAP expression. (A) SLAMF1 protein (Western blot) and SLAMF1 mRNA expression (real-time qRT-PCR). (B) SAP protein expression (ratio to GAPDH; Western blot). (C) SAP mRNA expression (normalized to Gapdh; real-time qRT-PCR). Data shown are means ± SD, n = 6 samples/group. ^※※※p < 0.001 vs. control group or NC siRNA group; ^▲▲▲p < 0.001 vs. control group; ^▼▼▼p < 0.001 vs. DEHP exposure group or DEHP + NC siRNA group; ns = p > 0.05 vs. control group or NC siRNA group.

Figure 4. Influence of Slamf1 siRNA transfection (with DEHP exposure) on T_fh cell nuclear transcription factor expression. (A) Expression percentages of CXCR5⁺BCL-6⁺ and CXCR5⁺c-MAF⁺ T_fh cells gated on CD4⁺ T-cell populations (%) (flow cytometry). (B) Bcl-6 and c-MAF mRNA (normalized to Gapdh; real-time qRT-PCR). Data shown are means ± SD, n = 6 samples/group. ^▲▲p < 0.01, ^▲▲▲p < 0.001 vs. control group; ^▼▼▼p < 0.001 vs. DEHP exposure group or DEHP + NC siRNA group; ns = p > 0.05 vs. control group or NC siRNA group.

Figure 5. Influence of Slamf1 siRNA transfection (with DEHP exposure) on T_fh cell cytokine secretion. Levels of IL-21 and IL-4 in T_fh cell culture supernatants as measured by ELISA. Data shown are means (pg/ml) ± SD, n = 6 samples/group. ^▲▲p < 0.01, ^▲▲▲p < 0.001 vs. control group; ^▼▼▼p < 0.001 vs. DEHP exposure group or DEHP + NC siRNA group; ns = p > 0.05 vs. control group or NC siRNA group.