Fig 1: Micrographs of gallbladder in children with cholecystitis and in controls. (a) Note a moderate to numerous number of IL-1α positive epithelial cells along the positive inflammatory cells in the capillaries and subepithelial tissue. IL-1α IMH, ×200; (b) no IL-1α positive cells were detected in the control. IL-1α IMH, ×200; (c) a moderate number of IL-4 positive cells in the connective tissue. IL-4 IMH, ×200; (d) note numerous IL-4 positive epitheliocytes and a lack of IL-4 positive cells in the connective tissues of control. IL-4 IMH, ×200; (e) numerous IL-6 containing epitheliocytes in a patient. IL-6 IMH, ×200; (f) control samples with also numerous IL-6 positive epithelial cells. IL-6 IMH, ×200.
Fig 2: Correlation of IDO1 expression with SOCS3 expression in chorionic villi (A and B) and decidua (C and D) cultured with low concentrations of IL-6 (0–2 ng/ml) for 24 h, as measured by qRT-PCR. The correlation of the IL-6 concentration added to the cultures with the expression of SOCS3 and IDO1 mRNA in chorionic villi (A) and decidua (C) cultures and the correlation of IDO1 mRNA expression with SOCS3 mRNA expression in chorionic villi (B) and decidua (D) cultured with low concentrations of IL-6 (0–2 ng/ml) were evaluated and analysed. Correlation of IDO1 expression with SOCS3 expression in chorionic villi (E and F) and decidua (G and H) cultured with high concentration of IL-6 (2–100 ng/ml) for 24 h, as measured by qRT-PCR. The correlation of the IL-6 concentration added to the cultures with the expression of SOCS3 and IDO1 mRNA in cultured chorionic villi (E) and decidua (G) and the correlation of IDO1 mRNA expression with SOCS3 mRNA expression in chorionic villi (F) and decidua (H) cultured with high concentrations of IL-6 (2–100 ng/ml) were evaluated and analysed. The data are presented as the mean ± SE of three similar experiments. IDO1, indoleamine 2,3-dioxygenase; SOCS3, suppressors of cytokine signalling 3; IL-6, interleukin-6
Fig 3: The expression of IL-6, IDO1, SHP-1, SHP-2, STAT3, pSTAT3 and SOCS3 and the correlation of these expression levels. A-D The expression of IL-6, IDO1, SHP-1, SHP-2, STAT3, pSTAT3, and SOCS3 and the correlation of these expression levels in chorionic villi. A. The expression of IL-6, IDO1, SHP-1, SHP-2, STAT3, pSTAT3 and SOCS3 in chorionic villi, as measured by Western blotting analysis. B, C and D The correlation of protein expression levels of IL-6, IDO1, SHP-1, SHP-2, STAT3, pSTAT3, and SOCS3 in chorionic villi. Data represent a portion of each sample from thirty-eight healthy pregnant women. Labels 1–8 represent different samples from healthy pregnant women. B The correlation coefficients (r) of the protein expression levels of IL-6 and IDO1, SHP-1, SHP-2, STAT3, pSTAT3 and SOCS3 were 0.735, 0.72, 0.77, 0.8047, 0.767, and − 0.844, respectively; the P values for the correlation of the protein expression levels between IL-6 and IDO1, SHP-1, SHP-2, STAT3, pSTAT3, and SOCS3 were 0.0377, 0.0400, 0.0241, 0.0160, 0.0262, and 0.0083, respectively. C The correlation coefficients (r) of the protein expression levels of IDO1 and SHP-1, SHP-2, STAT3, pSTAT3 and SOCS3 were 0.719, 0.7746, 0.78, 0.7147 and − 0.8923, respectively; the P values for correlation of the protein expression levels of IDO1 and SHP-1, SHP-2, STAT3, pSTAT3, and SOCS3 were 0.0443, 0.0240, 0.0198, 0.0463, and 0.0029, respectively. D The correlation of the protein expression levels of SOCS3 and SHP-1/2 was − 0.824/− 0.8773, respectively; the P value for the correlation of the protein expression levels of SOCS3 and SHP-1/2 was 0.0117/0.0042, respectively. E-H The expression of IL-6, IDO1, SHP-1, SHP-2, STAT3, pSTAT3, and SOCS3 and the correlation of these expression levels in decidua. E The expression of IL-6, IDO1, SHP-1, SHP-2, STAT3, pSTAT3 and SOCS3 in the decidua, as measured by Western blotting analysis. F, G and H The correlation of the protein expression levels of IL-6, IDO1, SHP-1, SHP-2, STAT3, pSTAT3 and SOCS3 in decidua. The data represent a portion of each sample from thirty-eight healthy pregnant women. Labels 1–8 represent different samples from healthy pregnant women. F The correlation coefficients (r) of the protein expression levels of IL-6 and IDO1, SHP-1, SHP-2, STAT3, pSTAT3, and SOCS3 were 0.8914, 0.8947, 0.8846, 0.8694, 0.8949, and − 0.7950, respectively; the P values for the correlation of the protein expression levels of IL-6 and IDO1, SHP-1, SHP-2, STAT3, pSTAT3 and SOCS3 were 0.0029, 0.0027, 0.0035, 0.0050, 0.0027 and 0.0183, respectively. G The correlation coefficients (r) of the protein expression levels of IDO1 and SHP-1, SHP-2, STAT3, pSTAT3 and SOCS3 were 0.834, 0.9890, 0.7359, 0.8527, and − 0.7896, respectively; the P values for correlation of the protein expression levels of IDO1 and SHP-1, SHP-2, STAT3, pSTAT3, and SOCS3 were 0.010, < 0.0001, 0.0374, 0.0071, and 0.0198, respectively. H The correlation of the protein expression levels of SOCS3 and SHP-1/2 was − 0.712/− 0.7665, respectively; the P value for correlation of the protein expression levels of SOCS3 and SHP-1/2 was 0.0451/0.0265, respectively. The greyscale ratios of IL-6 and IDO1, SOCS3, SHP-1, SHP-2, STAT3 and pSTAT3 and the expression levels of IL-6 and IDO1, SOCS3, SHP-1/2, STAT3 and pSTAT3 were assessed based on the ratios of the greyscale value of the IL-6 and IDO1, SOCS3, SHP-1, SHP-2, STAT3 and pSTAT3 bands to that of the GAPDH band. IDO1, indoleamine 2,3-dioxygenase; SOCS3, suppressors of cytokine signalling 3; STAT3, signal transducer and activator of transcription 3; pSTAT3, phosphorylated STAT3; SHP-1/2, protein tyrosine phosphatase-1/2; IL-6, interleukin-6. GAPDH, glyceraldehyde 3-phosphate dehydrogenase
Fig 4: A-D The expression of IDO1, SHP-1, SHP-2, STAT3, pSTAT3 and SOCS3 in chorionic villi cultured with a low concentration of IL-6 (0–2 ng/ml) for 24 h and the correlation of these expression levels, as measured by Western blotting analysis. The expression of IDO1, SHP-1, SHP-2, STAT3, pSTAT3, and SOCS3 (A), the correlation of the IL-6 concentration added to chorionic villi cultures with the expression of IDO1, SHP-1, SHP-2, STAT3, pSTAT3, and SOCS3 (B), the correlation of IDO1 expression with the expression of SHP-1, SHP-2, STAT3, pSTAT3, and SOCS3 (C), and the correlation of SOCS3 expression with the expression of SHP-1 and SHP-2 (D) in chorionic villi cultured with low concentrations of IL-6 (0–2 ng/ml) for 24 h were evaluated and analysed. The data are presented as the mean ± SE of three similar experiments. B The correlation coefficients (r) between the IL-6 concentration added to chorionic villi cultures and the expression of IDO1, SHP-1, SHP-2, STAT3, pSTAT3 and SOCS3 were 0.992, 0.988, 0.951, 0.995, 0.962, and 0.9458, respectively; the P values for the correlations of the levels of IL-6 and IDO1, SHP-1, SHP-2, STAT3, pSTAT3 and SOCS3 were 0.0175, 0.0427, 0.0445, 0.0426, 0.0412 and 0.0447, respectively. C The correlation coefficients (r) of the protein expression levels of IDO1 and SHP-1, SHP-2, STAT3, pSTAT3 and SOCS3 were 0.9824, 0.9906, 0.9911, 0.9511 and 0.9327, respectively; the P value for correlation of the protein expression levels of IDO1 and SHP-1, SHP-2, STAT3, pSTAT3 and SOCS3 were 0.037, 0.0427, 0.0401, 0.0477, and 0.0413, respectively. D The correlation of the protein expression levels of SOCS3 and SHP-1/2 was 0.9883 and 0.9730, respectively; the P value for correlation of the protein expression levels of SOCS3 and SHP-1/2 was 0.0421 and 0.0352, respectively. E-H The expression of IDO1, SHP-1, SHP-2, STAT3, pSTAT3, and SOCS3 in decidua cultured with a low concentration of IL-6 (0–2 ng/ml) for 24 h and the correlations of these expression levels, as measured by Western blotting analysis. The expression of IDO1, SHP-1, SHP-2, STAT3, pSTAT3 and SOCS3 (E), the correlation of the IL-6 concentration added to the decidua cultures with the expression of IDO1, SHP-1, SHP-2, STAT3, pSTAT3 and SOCS3 (F), the correlation of IDO1 expression with the expression of SHP-1, SHP-2, STAT3, pSTAT3, and SOCS3 (G), and the correlation of SOCS3 expression with the expression of SHP-1 and SHP-2 (H) in decidua cultured with low concentrations of IL-6 (0–2 ng/ml) for 24 h were evaluated and analysed. The data are presented as the mean ± SE of three similar experiments. F The correlation coefficients (r) between the IL-6 concentration added to the decidua cultures and the expression of IDO1, SHP-1, SHP-2, STAT3, pSTAT3, and SOCS3 were 0.950, 0.951, 0.980, 0.950, 0.9345, and 0.991, respectively; the P values for the correlations of IL-6 expression and IDO1, SHP-1, SHP-2, STAT3, pSTAT3 and SOCS3 expression were 0.0452, 0.0446, 0.0285, 0.0450, 0.0412, and 0.0193, respectively. G The correlation coefficients (r) of the protein expression levels of IDO1 and SHP-1, SHP-2, STAT3, pSTAT3 and SOCS3 were 0.976, 0.9866, 0.9967, 0.987 and 0.989, respectively; the P value for correlation of the protein expression levels of IDO1 and SHP-1, SHP-2, STAT3, pSTAT3 and SOCS3 were 0.0007, 0.0437, 0.0124, 0.0002, and 0.0345, respectively. H The correlation of the protein expression levels of SOCS3 and SHP-1/2 was 0.973/9630, respectively; the P value for the correlation of the protein expression levels of SOCS3 and SHP-1/2 was 0.0331/0.0452. The greyscale ratios of IDO1, SOCS3, SHP-1, SHP-2, STAT3 and pSTAT3 and the expression levels of IDO1, SOCS3, SHP-1/2, STAT3 and pSTAT3 were assessed based on the ratios of the greyscale values of the IL-6 and IDO1, SOCS3, SHP-1, SHP-2, STAT3 and pSTAT3 bands to that of the GAPDH band. IDO1, indoleamine 2,3-dioxygenase; SOCS3, suppressors of cytokine signalling 3; STAT3, signal transducer and activator of transcription 3; pSTAT3, phosphorylated STAT3; SHP-1/2, protein tyrosine phosphatase-1/2; IL-6, interleukin-6; GAPDH, glyceraldehyde 3-phosphate dehydrogenase
Fig 5: Diagram illustrating the molecular mechanism involved in the induction of IDO1 by IL-6
Supplier Page from Abcam for Anti-IL-6 antibody