Fig 1: Correlations between co-differentially expressed proteins and clinical indicators (TBA, gestational weeks, AST, and ALT) in pregnant women. (A) The correlation analysis between TBA and expression levels of NECTIN1, SPINT1, ENPP2, ITIH3, and SERPIND1. (B) The correlation analysis between gestational weeks and expression levels of NECTIN1, SPINT1, ENPP2, ITIH3, and SERPIND1. (C) The correlation analysis between AST and expression levels of NECTIN1, SPINT1, ENPP2, ITIH3, and SERPIND1. (D) The correlation analysis between ALT and expression levels of NECTIN1, SPINT1, ENPP2, and SERPIND1. TBA, total bile acid; AST, aspartate transaminase; ALT, alanine transaminase.
Fig 2: Correlation analysis of co-differentially expressed protein levels between placental and plasma samples. (A) Correlation analysis of RPS7 between placental and plasma samples. (B) Correlation analysis of NECTIN3 between placental and plasma samples. (C) Correlation analysis of NECTIN1 between placental and plasma samples. (D) Correlation analysis of SPINT1 between placental and plasma samples. (E) Correlation analysis of SERPINA7 between placental and plasma samples. (F) Correlation analysis of CD320 between placental and plasma samples. (G) Correlation analysis of ITIH3 between placental and plasma samples. (H) Correlation analysis of ENPP2 between placental and plasma samples. (I) Correlation analysis of SERPIND1 between placental and plasma samples.
Fig 3: Correlation analysis between ELISA and sequencing results and the validation of plasma differential proteins using ELISA. (A–E) The correlation analysis of SPINT1, SERPINA7, NECTIN1, SERPIND1, and ITIH3 between ELISA and sequencing results. (F) NECTIN1 levels in plasma samples obtained from ICP and control groups (P < 0.0001). (G) SPINT1 levels in plasma samples obtained from ICP and control groups (P < 0.0001). (H) ITIH3 levels in plasma samples obtained from ICP and control groups (P = 0.0001). (I) ENPP2 levels in plasma samples obtained from ICP and control groups (P < 0.0001). (J) SERPIND1 levels in plasma samples obtained from ICP and control groups (P < 0.0001); (****P < 0.0001, ***P < 0.001). ELISA, enzyme-linked immunosorbent assay; ICP, intrahepatic cholestasis of pregnancy.
Fig 4: Validation of placenta differential proteins using IHC. (A) NECTIN1 levels in the placental samples obtained from ICP and control groups (P = 0.0022). (B) SPINT1 levels in the placental samples obtained from ICP and control groups (P < 0.0001). (C) CD320 levels in the placental samples obtained from ICP and control groups (P < 0.0001). (D) ITIH3 levels in the placental samples obtained from ICP and control groups (P < 0.0001). (E) ENPP2 levels in the placental samples obtained from ICP and control groups (P < 0.0001). (F) SERPIND1 levels in the placental samples obtained from ICP and control groups (P < 0.0001); (****P < 0.0001, **P < 0.01). (G–L) IHC staining for NECTIN1, SPINT1, CD320, ITIH3, ENPP2, and SERPIND1 in the placental samples obtained from ICP and control groups ( × 200). IHC, immunohistochemistry; ICP, intrahepatic cholestasis of pregnancy.
Fig 5: Diagnostic utility of single and combined secreted protein levels in maternal plasma for pregnant women with ICP. (A–E) The ROC curve of NECTIN1, SPINT1, ITIH3, and SERPIND1. (F–L) The ROC curve of SERPIND1/NECTIN1, SERPIND1/SPINT1, SERPIND1/ITIH3, SERPIND1/NECTIN1/SPINT1, SERPIND1/NECTIN1/ITIH3, SERPIND1/SPINT1/ITIH3, and SERPIND1/NECTIN1/SPINT1/ITIH3. ICP, intrahepatic cholestasis of pregnancy; ROC, receiver operator characteristics.
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