Anti pcna antibody

The immunohistochemical staining was performed on paraffin-embedded lung 5-µm-thick sections. Proliferating cells were detected using an anti-PCNA antibody (Sigma-Aldrich).

Tissue sections were stained by H&E staining and observed under the Leica DM3000 microscope (Leica, Germany) to measure aortic wall thickness (WT), wall cross-sectional area (WCSA), lumen diameter (ID), and wall/lumen ratio. Fibers and collagens were stained by Masson's trichrome staining and Sirius-red staining. Quantitative analyses were performed by ImageJ.
After deparaffinage, rehydration, and permeabilization in 1‰ Triton X-100, double-immunofluorescence staining was performed on paraffin sections. The VSMCs in the aorta were labeled by anti-α-SMA antibody (Sigma Aldrich, USA). The ratio of NOX4-positive VSMCs and the expression of SOD2 in aortic VSMCs were marked with anti-NOX4 antibody and anti-SOD2 antibody (Sigma Aldrich, USA), respectively. Proliferative VSMCs were labeled with anti-PCNA antibody (Sigma Aldrich, USA) and Ki67 antibody (Sigma Aldrich, USA), respectively.

Fin regenerates were fixed at 4 dpa in 4% PFA overnight and decalcified with 0.5 M EDTA for 24 h. Sample were embedded in paraffin and sectioned at 5 µm. Immunohistochemistry with anti-PCNA antibody (Sigma) was performed as described [83] . Percentage of PCNA positive nuclei over Hoechst positive nuclei was determined on three to four sections of four independent samples for each genotype.