Smyd2

Antibodies to Kim-1, p21 and H3K36me3 were purchased from Abcam Inc. (Cambridge, United Kingdom). Antibodies to cleaved Caspase 3, Cyclin D1, SMYD2, p-p53, p53, p-STAT3, STAT3, p-NFκBp65, and Tubulin were purchased from Cell Signaling Technology (Danvers, MA). Antibodies to PCNA, F4/80, MCP-1, ICAM-1 and TUNEL were purchased from Servicebio (Wuhan, China). Antibodies to NGAL was purchased from R and D SYSTEMS (Minneapolis,United States). Antibodies to GAPDH were purchased from Arigo Biolaboratories Corp. (Shanghai, China). Antibodies to BAX was purchased from Absin Bioscience Inc. (Shanghai, China). Antibodies to Histone-H3 was purchased from Merck (Darmstadt, Germany). Antibodies to NFκBp65 was purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Antibodies to BCL-2 was purchased from Proteintech Group Inc. (Chicargo, United States). SMYD2 siRNA and scramble siRNA were purchased from GenePharma Co.Ltd. (Shanghai, China). AZ505 was purchased from MedChemExpress (New Jersey, United States). Cisplatin and all other chemicals were purchased from Sigma (St. Louis, MO).

Differentiated H9c2 cells or HL-1 cells transfected with HA-SMYD2 WT or C13S, or appropriate siRNA were subjected to serum starvation for 12 h and treated with AMA (2 μg/mL) in DMEM (5 mM glucose) for 12 h. Alternatively, cells were treated with ARP 100 (10 µM) for 1 h before incubation with AMA. Cells were then lysed with a lysis buffer. Proteins in lysates were resolved by SDS-PAGE and transferred to PVDF membrane. The membrane was blocked and incubated with primary antibodies toward α-actinin (Abcam, Cat# ab9465) (1:500), Hsp90 (BD transduction, Cat# 610418) (1:1000), actin (Abcam, Cat# ab3280) (1:2000), HA-tag (Biolegend, Cat# 901502) (1:1000), cardiac heavy chain myosin (Abcam, ab50967) (1:200), troponin I (Cell Signaling, Cat# 4002 S) (1:1000), β-tubulin (Santa Cruz) (1:1000), SMYD2 (Cell Signaling, Cat# 9734) (1:1000), or MMP-2 (Cell Signaling, Cat#4022 S) (1:1000) diluted in a blocking buffer at 4 °C overnight. Proteins were then visualized by chemiluminescence using appropriate HRP-conjugated secondary antibodies. Supplementary Fig. 19 shows the uncropped or original blots corresponding to the panels displayed in the figures.