Journal: International journal of molecular sciences

Article Title: SPINT1 Expressed in Epithelial Cells of Choroid Plexus in Human and Mouse Brains.

doi: 10.3390/ijms26115130

Figure Lengend Snippet: Figure 1. Representative microphotographs of immunoreactivities for SPINT1 (GTX114793) in mice (A–C) and expression of Spint1 and Gapdh mRNAs in CP located in the lateral (LV) and fourth (4V) ventricles and small intestine (SI) (D). Arrows indicate clear or strong immunoreactivity for SPINT1 (A–C). Scale bars indicate 10 µm.

Article Snippet: Primary antibodies against SPINT1 (HAI-1) (1:500, rabbit polyclonal, Cat. No. GTX114793; Gene Tex) [21], SPINT1 (HAI-1) (1:200, mouse monoclonal, sc-137159, clone H-1; Santa Cruz Biotechnology, Dallas, TX, USA) [13], E-cadherin (1:2000, rabbit polyclonal, Cat. No. 20874-1-AP; ProteinTech Group, Chicago, IL, USA) [22,23], Smad-interacting protein 1 (SIP1) (1:200, rabbit polyclonal, C10133, Assay Biotechnology Company, Fremont, CA, USA), mitochondria (1:200, mouse monoclonal, 909-301-D79, Rockland, Limerick, PA, USA), and EEA1, a marker for endosomes (1:100, rabbit polyclonal, NBP1-30914, Novus, Centennial, CO, USA) [24] were used in this study.

Techniques: Expressing

Journal: International journal of molecular sciences

Article Title: SPINT1 Expressed in Epithelial Cells of Choroid Plexus in Human and Mouse Brains.

doi: 10.3390/ijms26115130

Figure Lengend Snippet: Figure 2. Representative microphotographs of immunoreactivities for SPINT1 (sc-137159) in humans (A–J). (A–J) are staining images for SPINT1 in No. 1–10 shown in Table 1, respectively. Arrows indicate clear or strong immunoreactivity for SPINT1 in epithelial cells with enlarged cytoplasm (A–J). Insets of (A–J) show strong immunoreactivities in the enlarged cells. Scale bars indicate 100 µm.

Article Snippet: Primary antibodies against SPINT1 (HAI-1) (1:500, rabbit polyclonal, Cat. No. GTX114793; Gene Tex) [21], SPINT1 (HAI-1) (1:200, mouse monoclonal, sc-137159, clone H-1; Santa Cruz Biotechnology, Dallas, TX, USA) [13], E-cadherin (1:2000, rabbit polyclonal, Cat. No. 20874-1-AP; ProteinTech Group, Chicago, IL, USA) [22,23], Smad-interacting protein 1 (SIP1) (1:200, rabbit polyclonal, C10133, Assay Biotechnology Company, Fremont, CA, USA), mitochondria (1:200, mouse monoclonal, 909-301-D79, Rockland, Limerick, PA, USA), and EEA1, a marker for endosomes (1:100, rabbit polyclonal, NBP1-30914, Novus, Centennial, CO, USA) [24] were used in this study.

Techniques: Staining

Journal: International journal of molecular sciences

Article Title: SPINT1 Expressed in Epithelial Cells of Choroid Plexus in Human and Mouse Brains.

doi: 10.3390/ijms26115130

Figure Lengend Snippet: Figure 3. Representative confocal microscopic images of epithelial cells of CP from cases No. 7 (A–F,J–L), 9 (G–I), and 10 (M–O). Immunostaining with the mouse anti-SPINT1 antibody (A,D,G,J,M) (visualized as green), rabbit anti-E-cadherin (B,E,H) or SIP1 (K,N) antibody (visualized as red), nuclear staining by TO-PRO-3 (visualized as blue), and merged images (C,F,I,L,O) are shown. Solid long arrows in images (A,B,D,E,J,K,M,N) show double immunoreactivity. Dotted long arrows present strong SPINT1 and weak E-cadherin or SIP1 expression (D,E,G,H,J,K,M,N), whereas dotted double short arrows show weak SPINT1 and strong E-cadherin or SIP1 expression (D,E,G,H,J,K,M,N). Scale bars indicate 10 µm.

Article Snippet: Primary antibodies against SPINT1 (HAI-1) (1:500, rabbit polyclonal, Cat. No. GTX114793; Gene Tex) [21], SPINT1 (HAI-1) (1:200, mouse monoclonal, sc-137159, clone H-1; Santa Cruz Biotechnology, Dallas, TX, USA) [13], E-cadherin (1:2000, rabbit polyclonal, Cat. No. 20874-1-AP; ProteinTech Group, Chicago, IL, USA) [22,23], Smad-interacting protein 1 (SIP1) (1:200, rabbit polyclonal, C10133, Assay Biotechnology Company, Fremont, CA, USA), mitochondria (1:200, mouse monoclonal, 909-301-D79, Rockland, Limerick, PA, USA), and EEA1, a marker for endosomes (1:100, rabbit polyclonal, NBP1-30914, Novus, Centennial, CO, USA) [24] were used in this study.

Techniques: Immunostaining, Staining, Expressing

Journal: Heliyon

Article Title: Secreted proteins in plasma and placenta as novel non-invasive biomarkers for intrahepatic cholestasis of pregnancy: A case-control study

doi: 10.1016/j.heliyon.2023.e21616

Figure Lengend Snippet: Associated co-expressed differential proteins in placenta and plasma among pregnant women with ICP and healthy pregnant women.

Article Snippet: The primary antibodies used in this study were anti -NECTIN1 (1:100 dilution, Cat. 24713-1-AP, ProteinTech), anti -SPINT1 (1:100 dilution, Cat. ab189511; Abcam), anti -SERPINA7 (1:300 dilution; Cat. 66454-1-Ig; ProteinTech), anti-CD320 (1:300 dilution; Cat. 10343-1-AP, ProteinTech), anti -ITIH3 (1:100 dilution, Cat. DF14157, Affinity), anti -ENPP2 (1:50 dilution, Cat. 14243-1-AP, ProteinTech), and anti -SERPIND1 (1:25 dilution, Cat. ab232859; Abcam, Cambridge, UK).

Techniques: Clinical Proteomics, Protease Inhibitor

Journal: Heliyon

Article Title: Secreted proteins in plasma and placenta as novel non-invasive biomarkers for intrahepatic cholestasis of pregnancy: A case-control study

doi: 10.1016/j.heliyon.2023.e21616

Figure Lengend Snippet: 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.

Article Snippet: The primary antibodies used in this study were anti -NECTIN1 (1:100 dilution, Cat. 24713-1-AP, ProteinTech), anti -SPINT1 (1:100 dilution, Cat. ab189511; Abcam), anti -SERPINA7 (1:300 dilution; Cat. 66454-1-Ig; ProteinTech), anti-CD320 (1:300 dilution; Cat. 10343-1-AP, ProteinTech), anti -ITIH3 (1:100 dilution, Cat. DF14157, Affinity), anti -ENPP2 (1:50 dilution, Cat. 14243-1-AP, ProteinTech), and anti -SERPIND1 (1:25 dilution, Cat. ab232859; Abcam, Cambridge, UK).

Techniques: Clinical Proteomics

Journal: Heliyon

Article Title: Secreted proteins in plasma and placenta as novel non-invasive biomarkers for intrahepatic cholestasis of pregnancy: A case-control study

doi: 10.1016/j.heliyon.2023.e21616

Figure Lengend Snippet: 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.

Article Snippet: The primary antibodies used in this study were anti -NECTIN1 (1:100 dilution, Cat. 24713-1-AP, ProteinTech), anti -SPINT1 (1:100 dilution, Cat. ab189511; Abcam), anti -SERPINA7 (1:300 dilution; Cat. 66454-1-Ig; ProteinTech), anti-CD320 (1:300 dilution; Cat. 10343-1-AP, ProteinTech), anti -ITIH3 (1:100 dilution, Cat. DF14157, Affinity), anti -ENPP2 (1:50 dilution, Cat. 14243-1-AP, ProteinTech), and anti -SERPIND1 (1:25 dilution, Cat. ab232859; Abcam, Cambridge, UK).

Techniques: Enzyme-linked Immunosorbent Assay, Sequencing, Biomarker Discovery, Clinical Proteomics, Control

Journal: Heliyon

Article Title: Secreted proteins in plasma and placenta as novel non-invasive biomarkers for intrahepatic cholestasis of pregnancy: A case-control study

doi: 10.1016/j.heliyon.2023.e21616

Figure Lengend Snippet: 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.

Article Snippet: The primary antibodies used in this study were anti -NECTIN1 (1:100 dilution, Cat. 24713-1-AP, ProteinTech), anti -SPINT1 (1:100 dilution, Cat. ab189511; Abcam), anti -SERPINA7 (1:300 dilution; Cat. 66454-1-Ig; ProteinTech), anti-CD320 (1:300 dilution; Cat. 10343-1-AP, ProteinTech), anti -ITIH3 (1:100 dilution, Cat. DF14157, Affinity), anti -ENPP2 (1:50 dilution, Cat. 14243-1-AP, ProteinTech), and anti -SERPIND1 (1:25 dilution, Cat. ab232859; Abcam, Cambridge, UK).

Techniques: Biomarker Discovery, Control, Immunohistochemistry

Journal: Heliyon

Article Title: Secreted proteins in plasma and placenta as novel non-invasive biomarkers for intrahepatic cholestasis of pregnancy: A case-control study

doi: 10.1016/j.heliyon.2023.e21616

Figure Lengend Snippet: 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.

Article Snippet: The primary antibodies used in this study were anti -NECTIN1 (1:100 dilution, Cat. 24713-1-AP, ProteinTech), anti -SPINT1 (1:100 dilution, Cat. ab189511; Abcam), anti -SERPINA7 (1:300 dilution; Cat. 66454-1-Ig; ProteinTech), anti-CD320 (1:300 dilution; Cat. 10343-1-AP, ProteinTech), anti -ITIH3 (1:100 dilution, Cat. DF14157, Affinity), anti -ENPP2 (1:50 dilution, Cat. 14243-1-AP, ProteinTech), and anti -SERPIND1 (1:25 dilution, Cat. ab232859; Abcam, Cambridge, UK).

Techniques: Expressing

Journal: Heliyon

Article Title: Secreted proteins in plasma and placenta as novel non-invasive biomarkers for intrahepatic cholestasis of pregnancy: A case-control study

doi: 10.1016/j.heliyon.2023.e21616

Figure Lengend Snippet: 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.

Article Snippet: The primary antibodies used in this study were anti -NECTIN1 (1:100 dilution, Cat. 24713-1-AP, ProteinTech), anti -SPINT1 (1:100 dilution, Cat. ab189511; Abcam), anti -SERPINA7 (1:300 dilution; Cat. 66454-1-Ig; ProteinTech), anti-CD320 (1:300 dilution; Cat. 10343-1-AP, ProteinTech), anti -ITIH3 (1:100 dilution, Cat. DF14157, Affinity), anti -ENPP2 (1:50 dilution, Cat. 14243-1-AP, ProteinTech), and anti -SERPIND1 (1:25 dilution, Cat. ab232859; Abcam, Cambridge, UK).

Techniques: Diagnostic Assay, Clinical Proteomics

Journal: Heliyon

Article Title: Secreted proteins in plasma and placenta as novel non-invasive biomarkers for intrahepatic cholestasis of pregnancy: A case-control study

doi: 10.1016/j.heliyon.2023.e21616

Figure Lengend Snippet: Diagnostic value analysis of plasma proteins in ICP.

Article Snippet: The primary antibodies used in this study were anti -NECTIN1 (1:100 dilution, Cat. 24713-1-AP, ProteinTech), anti -SPINT1 (1:100 dilution, Cat. ab189511; Abcam), anti -SERPINA7 (1:300 dilution; Cat. 66454-1-Ig; ProteinTech), anti-CD320 (1:300 dilution; Cat. 10343-1-AP, ProteinTech), anti -ITIH3 (1:100 dilution, Cat. DF14157, Affinity), anti -ENPP2 (1:50 dilution, Cat. 14243-1-AP, ProteinTech), and anti -SERPIND1 (1:25 dilution, Cat. ab232859; Abcam, Cambridge, UK).

Techniques: Diagnostic Assay, Clinical Proteomics

Journal: Heliyon

Article Title: Secreted proteins in plasma and placenta as novel non-invasive biomarkers for intrahepatic cholestasis of pregnancy: A case-control study

doi: 10.1016/j.heliyon.2023.e21616

Figure Lengend Snippet: Diagnostic utility of combined plasma proteins in ICP.

Article Snippet: The primary antibodies used in this study were anti -NECTIN1 (1:100 dilution, Cat. 24713-1-AP, ProteinTech), anti -SPINT1 (1:100 dilution, Cat. ab189511; Abcam), anti -SERPINA7 (1:300 dilution; Cat. 66454-1-Ig; ProteinTech), anti-CD320 (1:300 dilution; Cat. 10343-1-AP, ProteinTech), anti -ITIH3 (1:100 dilution, Cat. DF14157, Affinity), anti -ENPP2 (1:50 dilution, Cat. 14243-1-AP, ProteinTech), and anti -SERPIND1 (1:25 dilution, Cat. ab232859; Abcam, Cambridge, UK).

Techniques: Diagnostic Assay, Clinical Proteomics