Journal: The FASEB Journal

Article Title: SMYD2 Promotes Renal Tubular Cell Apoptosis and Chronic Kidney Disease Following Cisplatin Nephrotoxicity

doi: 10.1096/fj.202402703R

Figure Lengend Snippet: AZ505 mitigates renal injury and suppresses the upregulation of ECM proteins in the renal tissue of mice with cisplatin‐induced CKD. (A, B) Serum blood urea nitrogen (BUN) and serum creatinine (Scr) levels in AZ505‐treated and untreated cisplatin‐induced CKD mice ( n = 5). (C) Hematoxylin and eosin (H&E), Masson's trichrome, and periodic acid‐Schiff (PAS) staining of renal tissue sections from AZ505‐treated and untreated cisplatin‐induced CKD mice. Scale bar: 10 μm. (D, E) Western blot analysis of ECM protein expression in renal tissues from AZ505‐treated and untreated cisplatin‐induced CKD mice ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: DMEM/F12 medium, fetal bovine serum (FBS) were purchased from Invitrogen Gibco, USA; tryptic digest, penicillin–streptomycin mixture were purchased from Biological Industries, Israel; Cis‐platin (CIS), SMYD2 specific inhibitor AZ505, NF‐κB specific inhibitor BAY11‐7085, Streptavidin‐FITC were purchased from APExBIO, USA; Smart‐ECL Enhanced Luminescence Kit was purchased from Changzhou Tiandi Renhe Bio‐technology Co. Ltd.; Hematoxylin and Eosin (HE) kit, MASSON Trichrome Staining Improvement Kit, Glycogen Staining (Periodic Acid Schiff (PAS)) kit, BCA Protein Concentration Measurement Kit, CCK‐8 (Cell Counting Kit‐8, CCK‐8) kit, Streptavidin‐FITC were purchased from APExBIO, USA. high‐efficiency RIPA (Radio Immuno Precipitation Assay, RIPA) lysate, anti‐fluorescence attenuation sealer containing 4′,6‐diamidino‐2‐phenylindole (DAPI), Bromphenol Blue, DL‐Dithiothreitol (DTT), NP‐40 LYSIS BUFFER lysate, and protease inhibitor (Phenylmethanesulfonyl fluoride (PMSF)) were purchased from Beijing Soleilbao Technology Co. TB GreeTM Premix Ex TaqTM II (TLi RNaseH Plus) was purchased from Takara Bio, Japan; Hifair III 1st Strand cDNA Synthesis SuperMix for qPCR (gDNA digester plus) was purchased from Yi Sheng Biotechnology (Shanghai); TUNEL kit (ElabscienceOne‐step TUNEL In Situ Apoptosis Kit) was purchased from Wuhan Elabscience Biotechnology Co. LTD; Goat anti‐Mouse IgG (H + L), CY3 AffiniPure was purchased from Wuhan Pramerica Biotechnology Co. biom; LTL, Biotinylated was purchased from Vector Laboratories Inc. in the USA; Aquaporin 1 (AQP1) (sc‐25 287), Protein A/G PLUS‐Agarose was purchased from SANTA CRUZ biotechnology Co. SANTA CRUZ biotechnology Inc.

Techniques: Staining, Western Blot, Expressing

Journal: The FASEB Journal

Article Title: SMYD2 Promotes Renal Tubular Cell Apoptosis and Chronic Kidney Disease Following Cisplatin Nephrotoxicity

doi: 10.1096/fj.202402703R

Figure Lengend Snippet: Smyd2 tecKO Mitigates kidney injury in cisplatin‐induced CKD mice and suppresses ECM protein upregulation. (A–C) Immunofluorescence and Western blot analysis of SMYD2 expression and localization in renal tissues of Smyd2 tecKO mice, with β‐Actin as a loading control (scale bar: 50 μm; N = 3). (D) Schematic of cisplatin‐induced CKD model in Smyd2 tecKO mice. (E–G) Blood urea nitrogen (BUN), serum creatinine (Scr), and proteinuria levels in cisplatin‐treated Smyd2 tecKO mice and Smyd2 fl/fl mice ( n = 5). (H) H&E, Masson, and PAS staining of kidney sections. Scale bar: 10 μm. (I, J) Western blot analysis of renal ECM proteins (α‐SMA, fibronectin, collagen I) in cisplatin‐treated groups ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: DMEM/F12 medium, fetal bovine serum (FBS) were purchased from Invitrogen Gibco, USA; tryptic digest, penicillin–streptomycin mixture were purchased from Biological Industries, Israel; Cis‐platin (CIS), SMYD2 specific inhibitor AZ505, NF‐κB specific inhibitor BAY11‐7085, Streptavidin‐FITC were purchased from APExBIO, USA; Smart‐ECL Enhanced Luminescence Kit was purchased from Changzhou Tiandi Renhe Bio‐technology Co. Ltd.; Hematoxylin and Eosin (HE) kit, MASSON Trichrome Staining Improvement Kit, Glycogen Staining (Periodic Acid Schiff (PAS)) kit, BCA Protein Concentration Measurement Kit, CCK‐8 (Cell Counting Kit‐8, CCK‐8) kit, Streptavidin‐FITC were purchased from APExBIO, USA. high‐efficiency RIPA (Radio Immuno Precipitation Assay, RIPA) lysate, anti‐fluorescence attenuation sealer containing 4′,6‐diamidino‐2‐phenylindole (DAPI), Bromphenol Blue, DL‐Dithiothreitol (DTT), NP‐40 LYSIS BUFFER lysate, and protease inhibitor (Phenylmethanesulfonyl fluoride (PMSF)) were purchased from Beijing Soleilbao Technology Co. TB GreeTM Premix Ex TaqTM II (TLi RNaseH Plus) was purchased from Takara Bio, Japan; Hifair III 1st Strand cDNA Synthesis SuperMix for qPCR (gDNA digester plus) was purchased from Yi Sheng Biotechnology (Shanghai); TUNEL kit (ElabscienceOne‐step TUNEL In Situ Apoptosis Kit) was purchased from Wuhan Elabscience Biotechnology Co. LTD; Goat anti‐Mouse IgG (H + L), CY3 AffiniPure was purchased from Wuhan Pramerica Biotechnology Co. biom; LTL, Biotinylated was purchased from Vector Laboratories Inc. in the USA; Aquaporin 1 (AQP1) (sc‐25 287), Protein A/G PLUS‐Agarose was purchased from SANTA CRUZ biotechnology Co. SANTA CRUZ biotechnology Inc.

Techniques: Immunofluorescence, Western Blot, Expressing, Control, Staining