Fig 1: Western blot, RT-qPCR and immunohistochemistry of the three biomarkers (IL1R1, CASP4 and IGSF6). (A) The time curves of the three biomarkers at mRNA level on day 1, 3, 5 and 7 after SCI or sham surgery (n = 3 rats per group at each time point, values are the mean ± SD, *p < 0.05, **p < 0.01, ***p < 0.001, t-test). (B) RT-qPCR validation of the biomarkers expressing in peak: the three signatures were increased significantly in SCI group (n = 3 rats per group, values are the mean ± SD, *p < 0.05, **p < 0.01, ***p < 0.001, t-test). (C) Western blot validation of the biomarkers as represented by their time curves at protein level (n = 3 rats per group, values are the mean ± SD, *p < 0.05, t-test). (D) Histomorphologically, the injured spinal cord tissue exhibited organizational abnormalities as compared with the normal control (overall scale bars = 200µm, regional-scale bars = 100µm). (E) Immunohistochemical validation of the biomarkers: the markers were found to be expressed in both gray matter and white matter (Overall scale bars = 200µm, regional-scale bars = 100µm, the arrows point to the positioning of the marker).
Fig 2: Acquisition and validation of candidate diagnostic markers. (A) LASSO regression showed the binomial deviation of different quantitative variables. The lower horizontal coordinate indicates the value of the lasso regression model parameter ?, the upper horizontal coordinate indicates the number of coefficients in the corresponding model, and the vertical coordinate indicates the error of the model. There are two dashed lines, the line with the lowest error on the left and the line with fewer features on the right. (B) Coefficient of LASSO regression model. As the ? value changes, the later the coefficient is compressed to zero the more important the variable is. (C) Boruta algorithm to screen diagnostic biomarkers. (D) Box violin maps of CASP4, IGSF6 and IL1R1 indicate that the differential expression in SCI group was significantly different from that in the normal group. (E) The ROC curves of the diagnostic efficacy of CASP4 (AUC=0.883), IGSF6 (AUC=0.907) and IL1R1 (AUC=0.942), the area under which was used to evaluate the effectiveness.
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