Fig 1: A representative Western blot of fibronectin, type I collagen, a-SMA, and CTGF protein in control (Con), Con + sRAGE, HMGB1 (10 µg/mL), and HMGB1+sRAGE (1 µg/mL) groups (Representative of six blots). Compared to Con cells, there were 2.1-, 1.9-, 4.2-, and 1.8-fold increases in fibronectin, type I collagen, a-SMA, and CTGF protein expression, respectively, in HMBG1-stimulated NRK-52E cells, and these changes were significantly abrogated by sRAGE treatment. *; p < 0.05 vs. Con and Con + sRAGE groups, #; p < 0.05 vs. HMGB1 group, **; p < 0.01 vs. Con and Con + sRAGE groups, ##; p < 0.01 vs. HMGB1 group. Each experiment was repeated six times independently with similar results.
Fig 2: A representative Western blot of NF-?B p65 protein in control (Con), Con + sRAGE, HMGB1 (10 µg/mL), and HMGB1+sRAGE (1 µg/mL) groups (Representative of six blots). Compared to Con cells, there was a significant decrease in NF-?B p65 protein expression in the cytosolic fraction by 51.2% and a significant 1.6-fold increase in the nuclear fraction of HMBG1-stimulated NRK-52E cells, and these changes were significantly attenuated by sRAGE treatment. *; p < 0.05 vs. Con and Con + sRAGE groups, #; p < 0.05 vs. HMGB1 group. Each experiment was repeated six times independently with similar results.
Fig 3: A representative Western blot of RAGE, phospho-ERK/ERK, phospho-p38/p38, phospho-JNK/JNK, and MyD88 protein in control (Con), Con + sRAGE, HMGB1 (10 µg/mL), and HMGB1+sRAGE (1 µg/mL) groups (Representative of six blots). Compared to Con cells, there were 5.3-, 2.6-, 1.8-, 1.8-, and 3.4-fold increases in RAGE, phospho-ERK/ERK, phospho-JNK/JNK, phospho-p38/p38, and MyD88 protein expression, respectively, in HMBG1-stimulated NRK-52E cells, and sRAGE significantly ameliorated these increases. *; p < 0.05 vs. Con and Con + sRAGE groups, #; p < 0.05 vs. HMGB1 group. Each experiment was repeated six times independently with similar results.
Fig 4: A representative Western blot (Representative of five blots) and immunohistochemical staining of HMGB1 and RAGE protein in the left kidney of sham-operated (Con), Con + sRAGE, UUO, and UUO + sRAGE rats. Compared to Con rats, there were 17.4- and 7.2-fold increases in HMGB1 and RAGE protein expression, respectively, in the left kidney of UUO rats treated with diluent (UUO), and administration of sRAGE significantly inhibited these increases in UUO rats (A). Compared to Con and Con + sRAGE rats, tubulointerstitial HMGB1 (B) and RAGE (C) protein expression were significantly increased in UUO rats treated with diluent, and these increases were significantly abrogated by sRAGE treatment. The significant increases in IHC staining scores for HMGB1 and RAGE within the tubulointerstitium of UUO rats were significantly mitigated in the UUO rats treated with sRAGE (D). *; p < 0.05 vs. Con and Con + sRAGE groups, **; p < 0.01 vs. Con and Con + sRAGE groups, #; p < 0.05 vs. UUO group, ##; p < 0.01 vs. UUO group. Each experiment (A–C) was repeated six times independently with similar results.
Fig 5: Serum HMGB1 levels measured by enzyme-linked immunosorbent assay of sham-operated (Con), UUO, and UUO + sRAGE rats. Compared to Con rats, serum concentration of HMGB1 was significantly higher in the UUO rats. sRAGE treatment decreased serum HMGB1 level slightly but it was not statistically significant. **; p < 0.01 vs. Con group.
Supplier Page from Novus Biologicals, a Bio-Techne Brand for Rat HMGB1/HMG-1 ELISA Kit (Colorimetric)