Cell Lysis and Western Blot Analysis

Cell lysis and Western blot analysis were performed as described previously [40 (link)], using the following antibodies: Rabbit polyclonal anti-SIK2 (Cell Signaling, Danvers, MA, USA), rabbit polyclonal anti-SIK2-pS385 (Kinexus, Sydney, Australia), rabbit polyclonal anti-AKT (Cell Signaling), rabbit polyclonal anti-AKT-pS473 (Cell Signaling), mouse monoclonal anti-β-actin (Sigma-Aldrich), rabbit polyclonal-anti KIF18B (ThermoFisher scientific, Waltham, MA, USA), rabbit polyclonal anti-EB1 (Abcam, Cambridge, UK), mouse monoclonal anti-cyclin B1 (Santa Cruz Biotechnology, Dallas, TX, USA), mouse monoclonal anti-Histone 3-pS10 (ThermoFisher scientific), mouse monoclonal anti-PLK1 (Santa Cruz Biotechnology), mouse monoclonal anti-vinculin (Santa Cruz Biotech), mouse monoclonal anti-P53 (Santa Cruz Biotech), rabbit polyclonal anti-P21 (cell signaling), rabbit polyclonal anti-BCL-xL (Santa Cruz Biotechnology), rabbit polyclonal anti-Cyclin D (Santa Cruz Biotechnology).
Reagents were purchased from the following sources: Paclitaxel (T7402) Sigma-Aldrich, propidium iodide (440300250) Acros Organics (Fair Lawn, NJ, USA), RNase A (1007885) Qiagen (Hilden, Germany), PE Annexin V (556421) and 7AAD (21-68981E) BD Biosciences, Caspase-Glo^® 3/7 Assay system (P1781) Promega, RO3306 (S7747) Selleckchem (Houston, TX, USA).

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Raab M., Rak M., Tesch R., Gasimli K., Becker S., Knapp S., Strebhardt K, & Sanhaji M. (2021). The Small-Molecule Inhibitor MRIA9 Reveals Novel Insights into the Cell Cycle Roles of SIK2 in Ovarian Cancer Cells. Cancers, 13(15), 3658.

Publication 2021

rabbit polyclonal anti-AKT mouse monoclonal anti-β-actin mouse monoclonal anti-cyclin B1 mouse monoclonal anti-PLK1 mouse monoclonal anti-vinculin mouse monoclonal anti-P53 rabbit polyclonal anti-P21 rabbit polyclonal anti-BCL-xL Paclitaxel (T7402) propidium iodide RNase A PE Annexin V (556421) and 7AAD (21-68981E)

7aad Actin Annexin v Antibodies Assay Caspase 7 Cell Cyclin Cyclin b1 Histone Mouse Paclitaxel Plk1 Promega Propidium iodide Rabbit Rnase Ro3306 Vinculin Western blot analysis

Corresponding Organization :

Other organizations : Goethe University Frankfurt

Top 5 similar protocols

Variable analysis

independent variables

Paclitaxel
RO3306

dependent variables

SIK2 expression
SIK2-pS385 expression
AKT expression
AKT-pS473 expression
β-actin expression
KIF18B expression
EB1 expression
Cyclin B1 expression
Histone 3-pS10 expression
PLK1 expression
Vinculin expression
P53 expression
P21 expression
BCL-xL expression
Cyclin D expression
Cell cycle progression (via propidium iodide and 7AAD staining)
Apoptosis (via PE Annexin V and 7AAD staining)
Caspase 3/7 activity

control variables

Cell lysis and Western blot analysis procedures (as described previously in reference 40)

positive controls

Not specified

negative controls

Not specified

Annotations

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