Ab205531

Antibodies raised against the following proteins were used at the indicated concentrations for western blotting (WB): rabbit anti-FLAG antibody (Sigma, F7425; 1:5000 for WB); rabbit anti-GRSF1 antibody (Abcam, ab205531; 1:1000 for WB); rabbit anti-Fibrinogen alpha chain antibody (Abcam, ab92572; 1:1000 for WB); rabbit anti-Fibrinogen beta chain antibody (Abcam, ab189490; 1:1000 for WB); mouse anti-Fibrinogen gamma chain antibody (Abcam, ab119948; 1:1000 for WB); rabbit anti-β-actin antibody (ABclonal, AC026; 1:10000 for WB); mouse anti-GAPDH antibody (ABclonal, AC002; 1:10000 for WB).

The western blot assay was performed as mentioned in previous studies [47 (link)]. The cells were seeded into 60 mm dishes at 2 × 10⁶ cells/dish. Protein extracted from cardiac tissues and cells was analyzed by the BCA Protein Assay Kit. SDS-PAGE (Sodium Dodecyl Sulfate—Polyacrylamide Gel Electrophoresis) was used to separate equal amounts of protein, which were then transferred to PVDF membranes. Membranes were incubated with primary antibodies overnight at 4 °C after being blocked with 5% milk. The primary antibodies used included TfR1 (A5865; ABclonal Technology, Woburn, MA, USA), ferritin heavy chain1 (FTH1, 3998; CST, Danvers, MA, USA), GPX4 (ab125066; Abcam, Cambridge, UK), Grsf1(ab205531; Abcam), cleaved Caspase 3 (AF7022; Affinity, West Bridgford, UK), Caspase 3 (AF6311; Affinity), Bax (14796; CST), Bcl2 (ab196495; Abcam), p38 MAPK (14451; CST), phosphop p38 MAPK (p-p38 MAPK;4631; CST), JNK (9252; CST), and phosphop JNK (p-JNK; 4668; CST). The blots were then incubated for 1.5 h with the corresponding secondary antibodies (ab7097; Abcam) and visualized using enhanced chemiluminescence (ECL) and the Bio-Rad imaging system. The protein signals were normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (ab181602; Abcam). The intensity of protein expression was quantified using ImageJ software.

Anti-peroxisome proliferator-activated receptor γ coactivator 1 alpha (PCG-1α) antibody (ab191838), anti-nuclear respiratory factor 1 (NRF1) antibody (ab175932), anti-mitochondrial transcription factor A (TFAM) antibody (ab176558), anti-ferritin heavy chain 1 (FTH1) antibody (ab65080), anti-transferrin receptor (ab269513), anti-glutathione peroxidase 4 antibody (ab125066) and anti-Grsf1 (ab205531) were purchased from Abcam. Rifampicin (R817237), isoniazid (I811711), and silybin (S817883) were purchased from Macklin Inc.