Journal: Journal of Cell Communication and Signaling

Article Title: Regulation of phosphatase and tensin homolog by complement component 5a (C5a) and its receptor (C5aR1) in lupus nephritis: A novel therapeutic target

doi: 10.1002/ccs3.70055

Figure Lengend Snippet: Upregulation of C5a/C5aR1 promotes cytokine release and drives inflammation in LN mice. (A) Schematic diagram of the animal model induction process; (B) Mouse cytokine antibody array Panel (A) showing increased C5a expression; (C) Western blot analysis of C5aR1 expression in mouse renal tissues, with β‐actin as the internal control ( n = 5); (D) Immunohistochemical analysis of C5aR1 expression in paraffin‐embedded kidney sections ( n = 5); (E) Analysis of IL‐1β and TNF‐α mRNA expression in mouse kidney tissues, with β‐actin as the internal control ( n = 5). (F) Representative H&E staining of paraffin‐embedded kidney sections ( n = 5); black arrows indicate inflammatory cell infiltration around renal tubules. Group comparisons were performed using a two‐tailed unpaired t ‐test, followed by LSD or Tukey post hoc tests to determine intergroup differences. Immunohistochemistry scores were analyzed using nonparametric tests. p > 0.05, not significant. **** p < 0.0001. H&E, hematoxylin and eosin; LN, lupus nephritis; LSD, least significant difference.

Article Snippet: Sections were incubated overnight at 4°C with rabbit primary antibodies, including C5aR1 (Proteintech), IL‐1β (Proteintech, Cat#16806‐1‐AP), monocyte chemoattractant protein‐1 (MCP‐1) (Abcam, Cat#ab25124), TNF‐α (Abcam, Cat#ab6671), and transforming growth factor‐β (TGF‐β) (Proteintech, Cat#21898‐1‐AP).

Techniques: Animal Model, Ab Array, Expressing, Western Blot, Control, Immunohistochemical staining, Staining, Two Tailed Test, Immunohistochemistry

Journal: Journal of Cell Communication and Signaling

Article Title: Regulation of phosphatase and tensin homolog by complement component 5a (C5a) and its receptor (C5aR1) in lupus nephritis: A novel therapeutic target

doi: 10.1002/ccs3.70055

Figure Lengend Snippet: C5a suppresses PTEN expression and enhances AKT pathway activation to promote inflammation. (A) Schematic diagram of the animal model procedure; (B) Schematic diagram of the in vitro cell experiment; (C) Protein‐protein interaction network illustrating key molecules linking C5a/C5aR11 with PTEN and the PI3K/AKT signaling pathway (confidence score = 0.15); (D) Western blot analysis of C5aR1 knockdown by three siRNAs in vitro (D1–D2) and by three shRNAs in kidney tissues in vivo (D3–D4), with GAPDH as loading control ( n = 3); (E) Western blot analysis of C5aR1 and PTEN expression after C5aR1 knockdown by siRNAs in vitro (E1–E3) and by shRNAs in kidney tissues in vivo (E4–E6), with GAPDH as loading control ( n = 3). (F–H) Results of the human/mouse AKT pathway phosphorylation antibody array C1 (RayBiotech) showing the expression of BAD, PRAS40, and PTEN. Group comparisons were performed using a two‐tailed unpaired t ‐test or one‐way ANOVA. p > 0.05, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. ANOVA, analysis of variance; BAD, Bcl‐2‐associated death promoter; PTEN, phosphatase and tensin homolog.

Article Snippet: Sections were incubated overnight at 4°C with rabbit primary antibodies, including C5aR1 (Proteintech), IL‐1β (Proteintech, Cat#16806‐1‐AP), monocyte chemoattractant protein‐1 (MCP‐1) (Abcam, Cat#ab25124), TNF‐α (Abcam, Cat#ab6671), and transforming growth factor‐β (TGF‐β) (Proteintech, Cat#21898‐1‐AP).

Techniques: Expressing, Activation Assay, Animal Model, In Vitro, Western Blot, Knockdown, In Vivo, Control, Phospho-proteomics, Ab Array, Two Tailed Test