Figure 4

Expression of p65 in T-cell subsets is altered during normal pregnancy compared to IUGR. (A) Gating strategy for flow cytometry analysis of cells expressing p65 in CD4/CD8⁺ T-cells. (B) Percentage of T-cell subsets expressing p65 was determined by flow cytometry in Th1, Th2, Th17, Treg cells, Tc1, and Tc2 from NP (n = 11), P (n = 12) and IUGR (n = 8). Box and whisker plots indicate median and IQR of cells expressing p65. *p < 0.05, **p < 0.01, ns = non- significant, determined by Kruskal–Wallis with Dunn’s multiple comparisons test due to small sample size. (C) Percentage of cells expressing p65 was compared between pro-inflammatory and anti-inflammatory CD4⁺ T-helper cell subsets as a ratio of Th1 and Th2, or Th17 and Treg, or CD8⁺ Tc1 and Tc2, from non-pregnant women (NP) (n = 11), normal pregnant women (P) (n = 12) and pregnant women complicated with IUGR (n = 8). Box and whisker plots indicate median and IQR of ratios. *p < 0.05, **p < 0.01, ns = non-significant, according to the Kruskal–Wallis. (D) Linear regression analysis was used to assess the relationship between regulation of transcription factors and p65 expression in CD4⁺ and CD8⁺ T-cells. p65 expression was correlated with CD4⁺ Tbet (Th1), GATA3 (Th2), RORγt (Th17), FOXP3 (Treg), CD8⁺ Tbet (Tc1), or CD8⁺ GATA3 (Tc2). Non-Pregnant (n = 11; blue); Pregnant (n = 12; green); IUGR (n = 8; red). Flow cytometry data were analysed using the FlowJo Software Version 10 (Becton Dickinson); website: https://www.flowjo.com/.