Fig. 11
From: Nicotine exacerbates diabetic nephropathy through upregulation of Grem1 expression
Nicotine increases the phosphorylation status of Smads, Ids, and Snail in mouse kidneys. A Western blot was performed to investigate the expression of p-Smad2/3 and p-Smad1/5/8 in the kidneys of Con, Nic, DM, and Nic + DM mice, and the representative gels are displayed. B The acquired images were used for data quantification, and the expression of p-Smad2/3 and p-Smad1/5/8 was normalized to β-actin. Results (mean ± SD) are representative of three independent samples. C, D Immunohistochemical staining of p-Smad2/3 and p-Smad1/5/8 in kidneys of Con and Nic, DM, Nic + DM mice. The scale bar is 20 μm. The average fluorescence intensity was measured per region of interest (ROI) for p-Smad2/3 and p-Smad1/5/8, and the results (mean ± SD) represent 10 randomly and blindly selected regions. E RNAs were extracted, and real-time PCR was performed to detect Id1, Id4, and Snail mRNA expression in the kidneys of Con and Nic, DM, and Nic + DM mice. Results (mean ± SD) are representative of three independent samples. * (p < 0.05) compared with Con group and # (p < 0.05) compared with DM group. Note: C, control; N, nicotine; D, diabetes mellitus; N + D, nicotine + diabetes mellitus