Monoclonal Anti-ACTA2 antibody produced in mouse

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	-Mouse (1:10,000)

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-Mouse (1:10,000)

Publication protocol

10–20 μg of the proteins were separated on SDS‐PAGE gels and transferred to a nitrocellulose membrane. The membranes were blocked with 10% nonfat milk for 60 min at room temperature, and then incubated with primary antibodies overnight at 4°C. The resources and dilutions of the primary antibodies are rabbit anti‐EZH2 monoclonal antibody (5246, Cell Signaling, 1:500), mouse anti‐ACTA2 (α‐SMA) monoclonal antibody (SAB1403519, SIGMA, 1:10,000), rabbit anti‐GAPDH polyclonal antibody (G9545, SIGMA, 1:10,000), rabbit anti‐p‐Smad2/3 monoclonal antibody (8828, Cell Signaling; 1:500), rabbit anti‐Smad2/3 monoclonal antibody (8685, Cell Signaling; 1:500), rabbit anti‐p‐Smad2 monoclonal antibody (3108, Cell Signaling; 1:500), rabbit anti‐p‐Smad3 monoclonal antibody (9520, Cell Signaling; 1:500), rabbit anti‐ACTB (β‐actin) polyclonal antibody (A2066, SIGMA, 1:10,000), and rabbit anti‐Lamin B1 monoclonal antibody (12586, Cell Signaling, 1:500). The membrane was washed using Tris‐buffered saline with Tween 20 (TTBS) and incubated with goat anti‐mouse or goat anti‐rabbit secondary antibodies (115‐035‐003 or 111‐035‐003, Jackson ImmunoResearch, West Grove, PA, 1:10000) for 1 h. The blots were then developed with enhanced chemiluminescence (ECL) reagents (PIERCE, Rockford, IL) and imaged with an Amersham Imager 600 (GE Healthcare Bio‐Sciences, Pittsburgh, PA).

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