Journal: Frontiers in Immunology

Article Title: To reveal biomarkers related to macrophage and lactic acid metabolism in renal fibrosis and explore their mechanisms

doi: 10.3389/fimmu.2025.1609903

Figure Lengend Snippet: Screening and diagnostic value of AGR3, CD74, and SYT11 in RF. (A) Least absolute shrinkage and selection operator (LASSO) regression analysis screening and cross-validation. (B) Boruta analysis screening results. Blue box: The minimum, average, and maximum Z-values of the shadow property; Red: rejection feature; Yellow: features to be confirmed; Green box: Confirmed feature. (C) Intersection of the two algorithms to obtain key feature genes. Pink represents genes confirmed by Lasso regression, and blue represents genes confirmed by Boruta as relevant. (D) Expression of key feature genes in the training set. (E) Expression of key feature genes in the validation set. (F) Receiver operating characteristic (ROC) analysis of candidate key genes in the training set. (G) ROC analysis of candidate key genes in the validation set.

Article Snippet: After incubation with 10% goat serum at room temperature for blocking, 100 μl of the working solution of CD74 (1:200) (Affinity, China), AGR2 + AGR3 (1:300) (since AGR3 and AGR2 are highly related homologous genes, we chose the AGR2 + AGR3 antibody for subsequent detection, Abcam, USA), and SYT11 (1:200) (Proteintech, China) primary antibodies were added to each tissue section and incubated at 4°C overnight.

Techniques: Diagnostic Assay, Selection, Biomarker Discovery, Expressing

Journal: Frontiers in Immunology

Article Title: To reveal biomarkers related to macrophage and lactic acid metabolism in renal fibrosis and explore their mechanisms

doi: 10.3389/fimmu.2025.1609903

Figure Lengend Snippet: Elucidating the biological mechanisms of biomarkers. (A) Gene set enrichment analysis (GSEA) of AGR3. (B) GSEA of CD74. (C) GSEA of SYT11.

Article Snippet: After incubation with 10% goat serum at room temperature for blocking, 100 μl of the working solution of CD74 (1:200) (Affinity, China), AGR2 + AGR3 (1:300) (since AGR3 and AGR2 are highly related homologous genes, we chose the AGR2 + AGR3 antibody for subsequent detection, Abcam, USA), and SYT11 (1:200) (Proteintech, China) primary antibodies were added to each tissue section and incubated at 4°C overnight.

Techniques:

Journal: Frontiers in Immunology

Article Title: To reveal biomarkers related to macrophage and lactic acid metabolism in renal fibrosis and explore their mechanisms

doi: 10.3389/fimmu.2025.1609903

Figure Lengend Snippet: Validation of key biomarker expression in animal experiments. (A) Representative kidneys of rats in the two groups at the time of kidney tissue collection. (B) Comparison of body weight and renal function between the two groups of rats. (C) HE, Masson and PAS pathological staining of kidneys in the two groups of rats. The black arrow indicates the area of this field of view in the renal tissue. (D) Immunohistochemical staining of AGR3, CD74 and SYT11 in kidney tissues of the two groups of rats. The black arrow indicates the area of this field of view in the renal tissue. (E) Immunofluorescence staining of AGR3, CD74 and SYT11 in kidney tissues of the two groups of rats. The white arrow indicates the area of this field of view in the renal tissue. (F) Western blot detection of differences in protein expression of AGR3, CD74 and SYT11 in kidney tissues of the two groups of rats. (B; mean ± SD, n=5; F, mean ± SD, n=3; Compared with the control group: ns, P ≥ 0.05; * P < 0.05; ** P < 0.01).

Article Snippet: After incubation with 10% goat serum at room temperature for blocking, 100 μl of the working solution of CD74 (1:200) (Affinity, China), AGR2 + AGR3 (1:300) (since AGR3 and AGR2 are highly related homologous genes, we chose the AGR2 + AGR3 antibody for subsequent detection, Abcam, USA), and SYT11 (1:200) (Proteintech, China) primary antibodies were added to each tissue section and incubated at 4°C overnight.

Techniques: Biomarker Discovery, Expressing, Comparison, Staining, Immunohistochemical staining, Immunofluorescence, Western Blot, Control