Fig 2: IFN-α is upregulated under hypoxic conditions in Kl−/− mice.(A) Western blot analysis demonstrating IFN-α expression in WT and Kl−/− mice. Protein levels were normalized to β-actin levels (n = 5 in each group). (B) Representative immunohistochemical staining images showing IFN-α expression in WT and Kl−/− mice. Values are mean ± SD. *P < 0.05, **P < 0.01. Bar = 100 μm.

Fig 3: Hypoxia enhances expression of RIG-I and IFN-α/β in a rat cell line of epithelial cells of renal tubules.NRK-52E cells were incubated under hypoxic conditions (1.0% O2) for 30, 60, 90, and 120 min. Cell lysates were subjected to western blot analysis using antibodies against RIG-I and IFN-α/β. Typical western blot analysis demonstrated the expression levels of RIG-I (A) and IFN-α/β (B and C). Graphs show the expression levels quantified by densitometry and normalized to α-tubulin (n = 5 in each group). Values are expressed as the mean ± SD. Statistical analysis was performed using ANOVA followed by Tukey’s post hoc test. *P < 0.05, **P < 0.01.

Fig 4: IFN-ß is increased under hypoxic conditions in Kl-/- mice.(A) Western blot analysis demonstrating IFN-ß expression in WT and Kl-/- mice. Protein levels were normalized to ß-actin levels (n = 5 in each group). (B) Representative immunohistochemical staining images showing IFN-ß expression in WT and Kl-/- mice. Values are mean ± SD. *P < 0.05, **P < 0.01. Bar = 100 µm.

Fig 5: RIG-I siRNA transfection attenuates expression of IFN-α/β in NRK-52E cells cultured under hypoxic conditions.NRK-52E cells were transfected with RIG-I siRNA or negative control siRNA. Cell lysates were subjected to western blot analysis using antibodies against RIG-I and IFN-α/β. Typical western blot analysis demonstrated the expression levels of RIG-I (A) and IFN-α/β (B and C). Graphs show the expression levels quantified by densitometry and normalized to α-tubulin (n = 5 in each group). Values are expressed as the mean ± SD. Statistical analysis was performed using ANOVA followed by Tukey’s post hoc test. *P < 0.05, **P < 0.01.