Journal: Renal Failure
Article Title: Sirt6 overexpression relieves ferroptosis and delays the progression of diabetic nephropathy via Nrf2/GPX4 pathway
doi: 10.1080/0886022x.2024.2377785
Figure Lengend Snippet: Figure 3. Expression of Sirt6 in renal tissues of db/m and db/db 16w group. (A–B) The proteins level of Sirt6, ACSL4, Nrf2, HO1, SOD2 and GPX4 were analyzed by Western blot in renal tissue of db/m and db/db group and shown using a histogram. (C)The mRNA levels of Sirt6, Nrf2, GPX4 were determined by real-time qPCR in renal tissues of db/m group and db/db group and shown using a histogram. (D) Immunohistochemical staining for Sitr6 in renal tissue of db/m group and db/db group. (E) The expression of Sirt6 in podocytes from db/db mice and db/m mice by immunofluorescence, WT-1 was used as podocyte marker. db/m: normal control group; db/db: 16w diabetic group; *p < 0.05, **p < 0.01 db/db vs db/m.
Article Snippet: Immunofluorescence assay Podocytes grown on cover slides were fixed with 4% formaldehyde for 30 min at room temperature, permeabilized 10 min with PbS containing 0.1% triton X-100, blocked for 10% fetal calf serum for 30 min at 37 °C. the samples were incubated with rabbit anti-Sirt6 polyclonal antibody (1:100, 13572-1-aP, Proteintech, Wuhan, China), rabbit anti-Nrf2 (1:50, 16396-1-aP, Proteintech), mouse anti-GPX4 (1:100, 67763-1-Ig, Proteintech), rabbit anti-Sod2 (1:50, 24127-1-aP, Proteintech), rabbit anti-Ho1 (1:200, 10701-1-aP, Proteintech) overnight at 4 °C, and then incubated with goat anti-rabbit or goat anti-mouse IgG H&L secondary antibody at 37 °C for 2h.
Techniques: Expressing, Western Blot, Immunohistochemical staining, Staining, Immunofluorescence, Marker, Control