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. 2023 May 8;13(9):2757–2773. doi: 10.7150/thno.81406

Figure 1.

Figure 1

HMGB1 is upregulated after RLDC treatment. C57BL/6 mice were injected weekly with 8 mg/kg cisplatin for four weeks and sacrificed for analysis one month later (RLDC group) or left untreated as controls (Con) to collect kidneys for analysis. (A) Representative immunoblots of fibronectin (FN), collagen-1 (COL-1), smooth muscle alpha-actin (α-SMA), and GAPDH (loading control) in kidney tissues. (B) Representative immunoblots of HMGB1 and GAPDH. (C) Densitometric analysis of HMGB1 immunoblots. (D) Quantitative analysis of serum HMGB1 by ELISA. (E) Representative images of immunohistochemical staining of HMGB1 in kidney tissues. The arrows point to nuclear staining of HMGB1 in control or nuclear and cytoplasmic staining in RLDC kidneys. Bar scale = 50 μm. (F) Quantification of HMGB1 positive areas on kidney sections. (G) Representative images of HMGB1 immunofluorescence (green) and Hoechst nuclear staining (blue) in kidney tissues. Scale bar = 50 μm; scale bar in the magnified images = 25 μm. (H) Quantitative analysis of HMGB1 staining in tubular cells. (I) qRT-PCR analysis of Hmgb1, Myd88, Tlr2, and Tlr4 mRNAs in control and RLDC kidneys. The expression of the target genes was normalized to GAPDH mRNA and expressed as fold change compared to control kidneys (Con). Quantitative data are expressed as mean ± SEM. N=5. *P < 0.05 vs. the control group (Con).