Fig 2: MS/MS fragment ion spectra of CRP and HSA from CRP-HSA crosslinked gel bands. Panel A: Representative MS/MS spectrum of peptide ESDTSYVSLK (m/z: 564.78; charge state: +2; mass error: 0.68 ppm) from CRP detected in CRP-HSA crosslinked gel band (band at ~ 75 kDa). Panel B: Representative MS/MS spectrum of peptide KVPQVSTPTLVEVSR (m/z: 820.47; charge state: 2+; mass error: 0.29 ppm) from HSA detected in CRP-HSA crosslinked gel band (band at ~ 75 kDa). Panel C: Representative MS/MS spectrum of peptide GYSIFSYATK (m/z: 568.79; charge state: 2+; mass error: 0.40 ppm) from CRP detected in CRP-HSA crosslinked gel band after reaction of photo-oxidized CRP and plasma (band at ~ 75 kDa). Panel D: Representative MS/MS spectrum of peptide YIC*ENQDSISSK (where * indicates carbamidomethylation of the Cys residues: + 57 Da; m/z: 722.33; charge state: 2+; mass error: 0.27 ppm) detected in CRP-plasma crosslinked gel band after reaction of photo-oxidized CRP and plasma (band at ~ 75 kDa). Red fragments correspond to the y ions, and blue fragments correspond to the b ions. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Fig 3: Downregulation of circ_0001578 induces inflammation in trophoblasts. (A) IL-1 levels were significantly higher in the culture medium treated with si-circRNA than in the si-NC group (P < 0.05). (B) IL-6 levels were significantly high in the culture medium treated with si-circRNA than in the si-NC group (P < 0.01). (C) IL-8 levels were significantly higher in the culture medium treated with si-circRNA than in the si-NC group (P < 0.01). (D) TNF-a levels were significantly higher in the culture medium treated with si-circRNA than in the si-NC group (P < 0.01). (E) CRP levels were significantly higher in the culture medium treated with si-circRNA than in the si-NC group (P < 0.001). (F) Expression levels of p-JNK and nuclear P65 followed by circ_0001578 knockdown detected by Western blotting. (G) Semiquantitative analysis of protein expression *P<0.05; **P<0.01; ***P<0.001.

Fig 4: miR-124 in ACI patients was negatively correlated with IL-6, IL-8, and CRP. The levels of IL-6, IL-8, and CRP were detected using ELISA kits. A, miR-124 was negatively correlated with IL-6. B, miR-124 was negatively correlated with IL-8. C, miR-124 was negatively correlated with CRP. Pearson correlation analysis was used for data analysis.

Fig 5: Formation of CRP-HSA crosslinks on exposure of purified CRP to 1O2 and subsequent reaction with HSA or plasma, and analysis by immunoblotting. Panels A and B: Formation of CRP-HSA crosslinks between oxidized CRP and HSA, as detected using an anti-CRP antibody and an anti-HSA antibody, respectively. Panels C and D: CRP-HSA crosslinks formed between oxidized CRP and plasma (2 mg protein mL-1, HSA: 0.7–1.2 g L-1), with detection by an anti-CRP antibody and an anti-HSA antibody, respectively; Lane 2 served as the positive control (reaction between oxidized CRP and purified HSA). Panel E: Optical density ratio (ODn min/OD0 min) of crosslinked bands from immunoblotting data presented in panels A–D (with n = 0, 5, 15 or 30 min): A and C are OD ratios from panels A and B, respectively; B and D are OD ratio from panels C and D, respectively. Statistical differences are indicated as follows: *P < 0.05 vs. lane 4 (panels A and B), #P < 0.05 vs. lane 5 (panels C and D). Error bars represent SD of at least three independent experiments.