Fig 1: Expression levels of SnoN and p-Smad2 in NRK-52E cells transfected with recombinant adenovirus. (A) Immunofluorescence analysis of the expression of smurf2 in NRK-52E cells infected with AD-smurf2. (B) RT-qPCR analysis was performed to detect the mRNA levels of smurf2, SnoN and Smad2, and western blot analysis was performed to detect the expression levels of smurf2, SnoN and p-Smad2 in NRK-52E cells infected with AD-smurf2. (C) Immunofluorescence analysis of the expression of smurf2 in NRK-52E cells treated with AD-shsmurf2. (D) RT-qPCR analysis was performed to detect the mRNA levels of smurf2, SnoN and Smad2, and western blot analysis was performed to detect the expression levels of smurf2, SnoN and p-Smad2 in NRK-52E cells infected with AD-shsmurf2. ***P<0.001, vs. control. SnoN, ski-related novel protein N; p-Smad2, phosphorylated small mothers against decapentaplegic; smurf2, Smad ubiquitination regulatory factor 2; AD, adenovirus; NC, negative control; RT-qPCR, reverse transcription-quantitative polymerase chain reaction.
Fig 2: Expression levels of smurf2, SnoN and p-Smad2 in NRK-52E cells incubated with low glucose medium. (A) Reverse transcription-quantitative polymerase chain reaction analysis was performed to detect the mRNA levels of smurf2, SnoN and Smad2 in NRK-52E cells incubated with low glucose medium at different time points. (B) Western blot analysis was performed to detect the expression levels of smurf2, SnoN, Smad2 and p-Smad2 in NRK-52E cells incubated with low glucose medium at different time points. *P<0.05 and ***P<0.001, vs. control. SnoN, ski-related novel protein N; p-Smad2, phosphorylated small mothers against decapentaplegic; smurf2, Smad ubiquitination regulatory factor 2.
Fig 3: Expression levels of smurf2, SnoN and p-Smad2 in DM rats injected with recombinant adenovirus. (A) RT-qPCR analysis was performed to detect the mRNA levels of smurf2, SnoN and Smad2, and western blot analysis was performed to detect the expression levels of smurf2, SnoN and p-Smad2 in animal models established via injection of recombinant adenovirus AD-smurf2 into DM rats. (B) Immunohistochemical staining showed the expression of smurf2 and SnoN in renal tubules of DM rats in different groups. (C) RT-qPCR analysis was performed to detect the mRNA levels of smurf2, SnoN and Smad2 and western blot analysis was performed to detect the expression levels of smurf2, SnoN and p-Smad2 in the DM rats injected with recombinant adenovirus AD-shsmurf. (D) Immunohistochemical staining showed the expression of smurf2 and SnoN in renal tubules of the DM rats in different groups. *P<0.05 and ***P<0.001, vs. control. DM, diabetes mellitus; SnoN, ski-related novel protein N; p-Smad2, phosphorylated small mothers against decapentaplegic; smurf2, Smad ubiquitination regulatory factor 2; AD, adenovirus; NC, negative control; RT-qPCR, reverse transcription-quantitative polymerase chain reaction.
Fig 4: Expression levels of smurf2, SnoN and p-Smad2 in NRK-52E cells treated with TGF-β1 and its inhibitor. (A) RT-qPCR analysis was performed to detect the mRNA levels of smurf2, SnoN and Smad2, and western blot analysis was performed to detect the expression levels of smurf2, SnoN and p-Smad2 in NRK-52E cells treated with TGF-β1 at different time points. (B) RT-qPCR analysis was performed to detect the mRNA levels of smurf2, SnoN and Smad2, and western blot analysis was performed to detect the expression levels of smurf2, SnoN and p-Smad2 in NRK-52E cells treated with TGF-β1 inhibitor. SnoN, ski-related novel protein N; p-Smad2, phosphorylated small mothers against decapentaplegic; smurf2, Smad ubiquitination regulatory factor 2; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; TGF-β1, transforming growth factor-β1.
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