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

Figure 6.

Figure 6

STAT1 regulates RLDC-induced cytoplasmic accumulation of HMGB1 in renal tubular cells. BUMPT cells were transfected with 50 nM STAT1 siRNA1 (siS1-1), siRNA2 (siS1-2), or negative control siRNA (siNC), and then subjected to 4 days of RLDC treatment or left untreated (Con). (A) Representative immunoblots of STAT1 and GAPDH (loading control). (B) Densitometry of STAT1. (C) Analysis of HMGB1 translocation by immunofluorescence and laser-scanning confocal microscopy; HMGB1 (green), nuclear staining with Hoechst (blue). Scale bar = 20 μm. (D) Quantitative analysis of nuclear and cytoplasmic HMGB1 signals and the nuclear-to-cytoplasmic ratio. (E) Representative immunoblots of nuclear and cytoplasmic HMGB1. Lamin B and GAPDH were used as internal controls for the nuclear and cytosolic factions, respectively. (F) Quantitative analysis of immunoblots of nuclear and cytoplasmic HMGB1. For calculation, the protein level in the control culture was arbitrarily set to 1, and the signals of other conditions were normalized with control to calculate fold changes. Data are expressed as mean ± SEM. N = 5. * P<0.05 vs. the control group (Con), # P < 0.05 vs. RLDC/siNC group.