Anti gamma h2a x antibody

First, MII oocytes were collected and washed with phosphate-buffered saline (PBS) containing 0.1% polyvinyl alcohol (PVA) and fixed with 4% paraformaldehyde (PFA) at room temperature (RT) for 30 min. Then, oocytes were permeabilized with 0.5% Triton X-100 for 20 min followed by a block with 1% BSA in PBS for 1 h. Next, oocytes were incubated with primary antibodies, anti-α-tubulin-fluorescein isothiocyanate (FITC) antibody (Invitrogen, United States, 1:300), phalloidin-FITC antibody (Sigma, United States, 1:300), and anti-gamma H2A.X antibody (Abcam, United Kingdom, 1:300) and FITC anti-mouse FR4 (Biolegend, United States, 1:100) overnight at 4°C. After that, oocytes were washed and incubated with appropriate secondary antibody (ZSGB-BIO, Beijing, China) for 1 h at RT. Oocytes were washed thoroughly and then stained with 10 μg/ml Hoechst 33,342 for 10 min. Finally, the oocytes were mounted on glass slides, and images were captured under a confocal laser scanning microscope (TCS-SP8MP, Leica, Germany).

Formalin‐fixed and paraffin‐embedded sections of the kidney were stained with Masson's trichrome. Fibrosis was detected using Masson trichrome staining. Kidney cryosections embedded in Tissue‐Tek O.C.T compound (4583, Sakura Finetek Japan) (5 μm‐thick slices) were fixed with acetone for 10 min. After washing three times with Dulbecco's phosphate buffered saline (PBS), the cells were incubated with 5% bovine serum albumin (BSA) (A3059, Sigma Aldrich) for 30 min, and subsequently with serum‐free protein block (X0909, DAKO) for 10 min. The slices were then incubated overnight at 4°C with primary antibodies, anti‐gamma H2A.X antibody (1:5000 dilution, ab11174, Abcam). The sections were incubated with Alexa Fluor 594 anti‐rabbit IgG secondary antibody (1:200 dilution, R37119, Thermo Fisher Scientific) for 30 min at room temperature. Nuclear staining was performed using bisbenzimide H 33342 trihydrochloride (B2261, Sigma Aldrich) for each sample. Images were acquired using an inverted fluorescent microscope, BZ‐X710 (Keyence), with the following filters: Texas Red with an excitation wavelength (Ex) of 560/40 nm, emission wavelength (Em) of 630/75 nm, and DAPI (Ex: 360/40 nm, Em: 460/50 nm).

Acetyl-Histone H4 (Lys8): Millipore, Billerica, MA; Cat # 17–10099
Anti-Histone H4 antibody: Millipore, Billerica, MA; Cat # 05-858R; clone 62-10C-2
Anti-hnRNPA2 antibody: Santa Cruz Biotechnology, Dallas, TX; Cat # sc-32316, clone # DP3B3
Anti- beta actin antibody: Santa Cruz Biotechnology, Dallas, TX; Cat # sc-47778, clone # C4
Anti-53BP1 antibody: Abcam, Cambridge, MA; Cat # ab36823
Anti-gamma H2A.X antibody: Abcam, Cambridge, MA; Cat # ab2893