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Blots were also washed three times with TBST 1× for 10 min and incubated overnight, in TBST containing 1% milk, with anti-DNMT1, DNMT3A, and DNMT3B monoclonal mouse antibodies (1 200), anti-TET1 polyclonal goat antibody (1 200), and anti-TET2 and anti-TET3 polyclonal rabbit antibodies (1 200).
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The blots were then washed in 1x TBST four times for 10 min.
The blots were then washed 3× for 10 min each time, with TBS-T.
The blots were subsequently washed with TBST and incubated with respective secondary antibodies.
Blots were again washed three times with TBS and then developed using enhanced chemiluminescence (ECL) kit.
The blots were then washed with TBS, and blocked with TBST.
Blots were stripped, washed, and reprobed for β-actin.
Blots were then washed three times at 50°C and autoradiographed.
The blots were again washed in TBS-T.
Blots were washed with TBST, followed by two washes in TBS without Tween prior to imaging.
Blots were washed for at least 30 mins in wash buffer (Tris/phosphate/saline/Tween).
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