Journal: The American journal of pathology

Article Title: CK2α/CSNK2A1 Phosphorylates SIRT6 and Is Involved in the Progression of Breast Carcinoma and Predicts Shorter Survival of Diagnosed Patients.

doi: 10.1016/j.ajpath.2016.08.007

Figure Lengend Snippet: Figure 2 Inhibition of CSNK2A1 decreased the proliferation and invasiveness of breast cancer cells. A and B: Treatment with the CSNK2 inhibitors, CX4945 (A) and emodin (B), significantly inhibited the growth of both MCF7 and T47D cells in a dose- and time-dependent manner. C: The knockdown of CSNK2A1 inhibited the proliferation of both MCF7 and T47D cells as indicated by MTT and colony-forming assays. D: The sub-G1 and G0/G1 population increased with the knockdown of CSNK2A1 in flow cytometric cell cycle analysis. E and F: The knockdown of CSNK2A1 significantly reduced cell migration (E) and invasion (F) in both MCF7 and T47D cell lines. *P < 0.05, **P < 0.01, and ***P < 0.001. CSNK2A1, casein kinase 2 a1.

Article Snippet: Two CSNK2 inhibitors, CX4945 (Santa Cruz Biotechnology, Santa Cruz, CA) and emodin (Sigma-Aldrich, St. Louis, MO), were used.

Techniques: Inhibition, Knockdown, Cell Cycle Assay, Migration

Journal: Cells

Article Title: Regulation of TMEM16A by CK2 and Its Role in Cellular Proliferation

doi: 10.3390/cells9051138

Figure Lengend Snippet: CK2 controls membrane expression of TMEM16A in CFBE airway epithelial cells. ( A ) Expression of double-tagged (eGFP and extracellular HA-tag) TMEM16A in CFBE airway epithelial cells. Membrane localized TMEM16A (Alexa647 positivity) was detected by an extracellular anti-HA-Alexa647-conjugated antibody. ( B , C ) RT-PCR and densitometric analysis indicating successful knockdown of CK2α’, #significant inhibition (unpaired t -test; p = 0.01). ( D , E ) Immunocytochemistry of TMEM16A expressed endogenously in CFBE cells. Membrane expression was reduced by knockdown of CK2α’, #significant inhibition (unpaired t -test; p = 0.000000002). Mean ± SEM. In parentheses are numbers of experiments.

Article Snippet: The CK2 inhibitors CX-4945 (silmitasertib) and TBB (4,5,6,7-Tetrabromobenzotriazole) were purchased from Cayman Chemicals and Sigma, respectively.

Techniques: Membrane, Expressing, Reverse Transcription Polymerase Chain Reaction, Knockdown, Inhibition, Immunocytochemistry

Journal: Cells

Article Title: Regulation of TMEM16A by CK2 and Its Role in Cellular Proliferation

doi: 10.3390/cells9051138

Figure Lengend Snippet: Inhibitors of CK2 inhibit TMEM16A in CFBE airway epithelial cells. ( A – F ) Whole cell current overlay recorded in patch clamp experiments and current/voltage relationships. ATP (100 µM) activated TMEM16A whole cell Cl − currents that were strongly inhibited by the CK2-inhibitors TBB (10 µM; #significant inhibition, unpaired t -test; p = 0.01, ( A , B )) and CX4945 (20 µM; #significant inhibition, unpaired t -test; p = 0.02; ( C , D )), and siRNA-knockdown of CK2α’ (#significant inhibition, unpaired t -test; p = 0.0001; ( E , F )). ( G , H ) Plasma membrane (PM) expression of endogenous TMEM16A in CFBE cells and inhibition of PM expression by the CK2-inhibitor CX4945 (#significant inhibition, unpaired t -test; p = 0.000000000007). Mean ± SEM #significant inhibition ( p < 0.05; unpaired t -test). In parentheses are numbers of experiments.

Article Snippet: The CK2 inhibitors CX-4945 (silmitasertib) and TBB (4,5,6,7-Tetrabromobenzotriazole) were purchased from Cayman Chemicals and Sigma, respectively.

Techniques: Patch Clamp, Inhibition, Knockdown, Clinical Proteomics, Membrane, Expressing

Journal: Cells

Article Title: Regulation of TMEM16A by CK2 and Its Role in Cellular Proliferation

doi: 10.3390/cells9051138

Figure Lengend Snippet: Role of CK2 for plasma membrane expression of TMEM16A in Cal33 head and neck cancer cells. ( A , B ) RT-PCR and densitometric analysis indicating successful knockdown of CK2α’ by siRNA for CK2α’ in Cal33 head and neck cancer cells (#significant inhibition, unpaired t -test; p = 0.01). Knockdown of CK2α’ did not inhibit transcription of TMEM16A. ( C , D ) Western blot analysis indicating successful knockdown of CK2α’ but unaffected expression of TMEM16A. ( E , F ) Plasma membrane (PM) expression of TMEM16A expressed endogenously in Cal33 cells and inhibition of PM expression by knockdown of CK2α’ (#significant inhibition, unpaired t -test; p = 0.00000002). ( G ) Current/voltage relationships of ATP-activated TMEM16A whole cells currents, indicating inhibition of TMEM16A by knockdown of CK2α’ (#significant inhibition, unpaired t -test; p = 0.01). Mean ± SEM. In parentheses are numbers of experiments.

Article Snippet: The CK2 inhibitors CX-4945 (silmitasertib) and TBB (4,5,6,7-Tetrabromobenzotriazole) were purchased from Cayman Chemicals and Sigma, respectively.

Techniques: Clinical Proteomics, Membrane, Expressing, Reverse Transcription Polymerase Chain Reaction, Knockdown, Inhibition, Western Blot

Journal: Cells

Article Title: Regulation of TMEM16A by CK2 and Its Role in Cellular Proliferation

doi: 10.3390/cells9051138

Figure Lengend Snippet: Inhibition of proliferation by knockdown of CK2α’ and TMEM16A. ( A ) Cell proliferation assessed in MTT assays and shown as absorbance. Both siRNA-knockdown of CK2α’ and TMEM16A inhibited cell proliferation (#significant inhibition, unpaired t -tests; p = 0.0001). Simultaneous knockdown of CK2α’ and TMEM16A had a more pronounced inhibitory effect on cell proliferation (#significant inhibition, unpaired t -test; p = 0.0015). ( B ) Inhibition of cell proliferation by the CK2-inhibitor CX4945 (20 µM) and additional inhibitory effect of TMEM16A-knockdown (#significant inhibition, unpaired t -test; p = 0.0001). Mean ± SEM. In parentheses are numbers of experiments.

Article Snippet: The CK2 inhibitors CX-4945 (silmitasertib) and TBB (4,5,6,7-Tetrabromobenzotriazole) were purchased from Cayman Chemicals and Sigma, respectively.

Techniques: Inhibition, Knockdown

Journal: Cells

Article Title: Regulation of TMEM16A by CK2 and Its Role in Cellular Proliferation

doi: 10.3390/cells9051138

Figure Lengend Snippet: Blockers of CK2 and TMEM16A inhibit proliferation of Cal33 and BHY head and neck cancer cells. ( A ) Blocking CK2 by CX4945 (20 µM) and blocking TMEM16A by niclosamide (0.5 µM) inhibited proliferation of Cal33 cells. Simultaneous application of both blockers had an additive effect (#significant inhibition, unpaired t -tests; p = 0.0001). ( B ) Enhanced cell proliferation of BHY cells induced by the TMEM16A-activator, Eact. Blocking CK2 by CX4945 (20 µM) and blocking TMEM16A by niclosamide (0.5 µM) inhibited proliferation of BHY cells. Simultaneous application of both blockers had an additive effect (#significant inhibition, unpaired t -tests; p = 0.000015). Mean ± SEM. In parentheses are numbers of experiments.

Article Snippet: The CK2 inhibitors CX-4945 (silmitasertib) and TBB (4,5,6,7-Tetrabromobenzotriazole) were purchased from Cayman Chemicals and Sigma, respectively.

Techniques: Blocking Assay, Inhibition

Journal: Cells

Article Title: Regulation of TMEM16A by CK2 and Its Role in Cellular Proliferation

doi: 10.3390/cells9051138

Figure Lengend Snippet: Blockers of CK2 and TMEM16A inhibit receptor-mediated Ca 2+ signaling. ( A , B ) Original recordings and summaries for basal and ATP-induced intracellular Ca 2+ concentrations in Cal33 cells. Increase of intracellular Ca 2+ by 10 and 100 µM ATP, respectively. Both CX4945 (20 µM; #significant inhibition, ANOVA; p = 0.0004) and niclosamide (1 µM; #significant inhibition, ANOVA; p = 0.0002) largely reduced ATP-induced Ca 2+ increase. Mean ± SEM. In parentheses are numbers of experiments.

Article Snippet: The CK2 inhibitors CX-4945 (silmitasertib) and TBB (4,5,6,7-Tetrabromobenzotriazole) were purchased from Cayman Chemicals and Sigma, respectively.

Techniques: Inhibition