Western Blot Analysis of Histone Deacetylases

RPMI culture medium was purchased from Corning (Corning, NY). Fetal bovine serum (FBS) was purchased from Sigma Chemical Co. (St. Louis, MO). Trypsin, pen/strep antibiotics, and puromycin were purchased from Gibco (Grand Island, NY). Trametinib (MEK inhibitor) was purchased from Selleckchem (Houston, TX). Antibodies for Western Blot: Anti-HDAC1 (#2062, from Cell Signaling Technology (Danvers, MA), anti-HDAC2 (#2540 CST), The anti-HDAC3 and anti-HDAC8 antibodies were described in refs. 17 (link), HDAC11(#58442, CST). Anti-HDAC6 (#H-300, sc-11420) was purchased from Santa Cruz Biotechnologies. Cleaved PARP (Asp214) (#D64E10) (#5625 CST), phospho LKB1 (#3482, CST), total LKB1 (#3050, CST). Anti-YAP1 antibody (#H00010413-M01, Abnova). Anti-Vinculin (#G8796) and anti-GAPDH (#V9131) were purchased from Millipore Sigma (Bedford, MA).

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Sriramareddy S.N., Faião-Flores F., Emmons M.F., Saha B., Chellappan S., Wyatt C., Smalley I., Licht J.D., Durante M.A., Harbour J.W, & Smalley K.S. (2022). HDAC11 activity contributes to MEK inhibitor escape in uveal melanoma. Cancer gene therapy, 29(12), 1840-1846.

Publication 2022

RPMI culture medium Fetal bovine serum (FBS) Trypsin pen/strep antibiotics puromycin Trametinib Anti-HDAC1 anti-HDAC2 Anti-HDAC6 Anti-YAP1 antibody Anti-Vinculin anti-GAPDH

Anti antibodies Anti antibody Antibiotics Antibodies Culture medium Fetal bovine serum Gapdh Hdac11 Lkb1 Puromycin Strep Trametinib Trypsin Vinculin Western blot Yap1

Corresponding Organization :

Other organizations : Moffitt Cancer Center, University of Florida Health, Sylvester Comprehensive Cancer Center, University of Miami

Top 5 similar protocols

Variable analysis

independent variables

Trametinib (MEK inhibitor)

dependent variables

HDAC1 expression
HDAC2 expression
HDAC3 expression
HDAC8 expression
HDAC11 expression
HDAC6 expression
Cleaved PARP (Asp214) expression
Phospho LKB1 expression
Total LKB1 expression
YAP1 expression

control variables

RPMI culture medium
Fetal bovine serum (FBS)
Trypsin
Pen/strep antibiotics
Puromycin
Anti-HDAC1 antibody
Anti-HDAC2 antibody
Anti-HDAC3 antibody
Anti-HDAC8 antibody
Anti-HDAC11 antibody
Anti-HDAC6 antibody
Anti-cleaved PARP antibody
Anti-phospho LKB1 antibody
Anti-total LKB1 antibody
Anti-YAP1 antibody
Anti-Vinculin antibody
Anti-GAPDH antibody

Annotations

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