Anti collageni ab34710

Anti-α-SMA (cs19245), anti-p-AMPKα (cs2535), anti-caspase-1 (cs24232) and anti-NOD-like receptor thermal protein domain associated protein 3 (NLRP3) (cs15101) were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-CollagenI (ab34710), anti-Tissue Inhibitors of Metalloproteinase 1 (TIMP1) (ab61224), anti-caspase-3 (ab179517), anti-beclin-1 (ab210498), anti-Bcl-2 (ab194583) antibodies and the horseradish peroxidase (HRP)-conjugated goat anti-rabbit (ab97051) were purchased from Abcam (Cambridge, MA, USA). Anti-CollagenI (14695-1-AP), anti-SIRT3 (10099-1-AP), anti-F4/80 (28463-1-AP), anti-caspase-6 (10198-1-AP), anti-caspase-9 (10380-1-AP) and anti-GAPDH (60004-1-Ig) antibodies were purchased from Proteintech (Wuhan, HubChina). Anti-Matrix metalloproteinase-13 (MMP-13) (sc515284) and anti-Interleukin (IL)-1R1 (sc393998) antibodies were purchased from Santa Cruz Biotechnology (Stanta Cruze, CA, USA). Anti-IL-1β (mab4012) antibodies were purchased from R&D Systems (Minneapolis, MN, USA). Alexa Fluor 488 Goat anti-Mouse IgG (A11034) and Alexa Fluor Plus 647 Goat anti-Rabbit IgG (A32727) were purchased from Beyotime Biotechnology (Shanghai, China).

The peri-infract regions of myocardial or CFs were lysed in RIPA buffer containing PMSF (Beyotime, Shanghai, China). After being homogenized and centrifuged (12 000 rpm, 10 min), the supernatant protein was transferred to another tube and subjected to standard BCA assay (Solarbio, Beijing, China). We separated 50 μg of proteins to SDS-PAGE on 12% polyacrylamide gels, transferred it to a PVDF microporous membrane (Millipore, Bedford, MA, USA), then blocked it with buffer containing TBST and 5% fat-free milk for 2 h at room temperature. The PVDF membrane was incubated in the primary antibodies (diluted by TBST with 0.1% tween) at 4°C overnight. After washing 3 times, the PVDF membrane was incubated with horseradish peroxidase (HRP)-conjugated anti-rabbit (1: 1000, Cell Signaling Technology, Inc., Danvers, MA, USA) for 2 h at room temperature. High-sensitivity chemiluminescence (Bio-Rad, Hercules, CA, USA) was used to detect the protein bands. The relative band densities were normalized to β-actin (1: 1000, Cell Signaling Technology, Inc., Danvers, MA, USA). The primary antibodies Anti-Collagen I (ab34710, 1: 1000) and Anti-Collagen III (ab7778, 1: 1000), purchased from Abcam (USA).