ck2β Search Results


antibodies against ck2 β  (Becton Dickinson)
90
Becton Dickinson antibodies against ck2 β
Antibodies Against Ck2 β, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rabbit monoclonal anti-ck2β antibody  (WuXi AppTec)
90
WuXi AppTec rabbit monoclonal anti-ck2β antibody
a Immunoblotting showing the expression pattern of <t>CK2β</t> in mouse oocytes at different developmental stages. A total 150 oocytes were collected after being cultured for 0, 4, 8, and 12 h, corresponding to GV, GVBD, MI, and MII stages, respectively. β-actin was detected as an internal control. b Immunohistochemistry analysis of the expression pattern of CK2β in primordial, primary, secondary, and tertiary follicles at PD21. c Immunohistochemistry detection of CK2β loss in oocytes of Ck2β fl/fl ;GCre + mice. Scale bar: 50 μm
Rabbit Monoclonal Anti Ck2β Antibody, supplied by WuXi AppTec, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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ck2α (#611610)  (Becton Dickinson)
90
Becton Dickinson ck2α (#611610)
a Immunoblotting showing the expression pattern of <t>CK2β</t> in mouse oocytes at different developmental stages. A total 150 oocytes were collected after being cultured for 0, 4, 8, and 12 h, corresponding to GV, GVBD, MI, and MII stages, respectively. β-actin was detected as an internal control. b Immunohistochemistry analysis of the expression pattern of CK2β in primordial, primary, secondary, and tertiary follicles at PD21. c Immunohistochemistry detection of CK2β loss in oocytes of Ck2β fl/fl ;GCre + mice. Scale bar: 50 μm
Ck2α (#611610), supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rrr-qaasnfkspvktir peptide corresponding to ck2β human sequence [203–215]  (MultiSynTech gmbh)
90
MultiSynTech gmbh rrr-qaasnfkspvktir peptide corresponding to ck2β human sequence [203–215]
a Immunoblotting showing the expression pattern of <t>CK2β</t> in mouse oocytes at different developmental stages. A total 150 oocytes were collected after being cultured for 0, 4, 8, and 12 h, corresponding to GV, GVBD, MI, and MII stages, respectively. β-actin was detected as an internal control. b Immunohistochemistry analysis of the expression pattern of CK2β in primordial, primary, secondary, and tertiary follicles at PD21. c Immunohistochemistry detection of CK2β loss in oocytes of Ck2β fl/fl ;GCre + mice. Scale bar: 50 μm
Rrr Qaasnfkspvktir Peptide Corresponding To Ck2β Human Sequence [203–215], supplied by MultiSynTech gmbh, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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recombinant human ck2β protein  (GeneTex)
90
GeneTex recombinant human ck2β protein
<t>CK2</t> interacts with NOXO1 in T84 colon epithelial cells under inflammatory conditions. ( A ) Immunoblots of NOXO1, CK2α/α′, CK2β, and β-actin in colon T84 cells stimulated with TNFα (5 ng/mL) or IL17 (50 ng/mL) individually or in combination for 24 hours at 37°C. Representative of 3 independent experiments. ( B ) ROS production was measured by chemiluminescence in T84 cells stimulated as in (A) . n = 3. ( C ) Immunoblots of CK2 α/α′ in NOXO1 IPP from T84 cells stimulated as in (A) . Representative of 4 independent experiments. ( D ) Densitometry analysis of CK2 α/α′ in NOXO1 IPP as in ( C ), normalized to CK2 α/α′expression in cell lysates; n = 4; mean ± SEM; one-way ANOVA with Tukey multiple comparisons test; ∗ P < .05. ( E ) Confocal microcopy of T84 cells co-stimulated or not with TNFα + IL17 for 24 hours at 37°C. NOXO1 ( green ), CK2 α/α′ ( red ), DAPI ( blue ). Scale bar: 10 μm. Representative of 6 independent experiments.
Recombinant Human Ck2β Protein, supplied by GeneTex, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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recombinant human ck2β protein - by Bioz Stars, 2026-03
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pcdna5-flag-ck2β (wt or kssr mutant  (Froggabio inc)
90
Froggabio inc pcdna5-flag-ck2β (wt or kssr mutant
<t>CK2</t> interacts with NOXO1 in T84 colon epithelial cells under inflammatory conditions. ( A ) Immunoblots of NOXO1, CK2α/α′, CK2β, and β-actin in colon T84 cells stimulated with TNFα (5 ng/mL) or IL17 (50 ng/mL) individually or in combination for 24 hours at 37°C. Representative of 3 independent experiments. ( B ) ROS production was measured by chemiluminescence in T84 cells stimulated as in (A) . n = 3. ( C ) Immunoblots of CK2 α/α′ in NOXO1 IPP from T84 cells stimulated as in (A) . Representative of 4 independent experiments. ( D ) Densitometry analysis of CK2 α/α′ in NOXO1 IPP as in ( C ), normalized to CK2 α/α′expression in cell lysates; n = 4; mean ± SEM; one-way ANOVA with Tukey multiple comparisons test; ∗ P < .05. ( E ) Confocal microcopy of T84 cells co-stimulated or not with TNFα + IL17 for 24 hours at 37°C. NOXO1 ( green ), CK2 α/α′ ( red ), DAPI ( blue ). Scale bar: 10 μm. Representative of 6 independent experiments.
Pcdna5 Flag Ck2β (Wt Or Kssr Mutant, supplied by Froggabio inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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30 nm ck2β specific sirna (sequence target: 5'-gccatggtgaagctctact-3' )  (SignaGen)
90
SignaGen 30 nm ck2β specific sirna (sequence target: 5'-gccatggtgaagctctact-3' )
(A) HK-2 control (shCV) and silenced cells (shCK2α and shCK2β) were analysed for the expression of each CK2 subunit (α,α’ or β) and for the phosphorylation state of the CK2 target Akt S129. The 1AD9 anti-CK2α antibody detects also CK2α’; the arrows indicate the migration of each catalytic isoform. (B) Detection of total and phosphorylated eIF2β(S2) using specific antibodies. β-actin was used as a loading control. (C) The phosphorylation of endogenous CK2 substrates were analysed in lysates of HK-2 control (shCV) and silenced cells (shCK2α and shCK2β) by Western blot using Phospho-(Ser/Thr) CK2 Substrate (P-S/T3-100) antibody. Arrows indicate the bands that are more affected by <t>CK2β</t> down-regulation, whose quantification is shown in the bar graph. Tubulin was used as a loading control. A representative Western blot is shown in each case, while quantification (means ± SEM) is shown in the bar graphs. n = 4 for panel (A) and (B), n = 3 for panel (C).
30 Nm Ck2β Specific Sirna (Sequence Target: 5' Gccatggtgaagctctact 3' ), supplied by SignaGen, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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ck2β-floxed mice  (Inserm Transfert)
90
Inserm Transfert ck2β-floxed mice
(A) HK-2 control (shCV) and silenced cells (shCK2α and shCK2β) were analysed for the expression of each CK2 subunit (α,α’ or β) and for the phosphorylation state of the CK2 target Akt S129. The 1AD9 anti-CK2α antibody detects also CK2α’; the arrows indicate the migration of each catalytic isoform. (B) Detection of total and phosphorylated eIF2β(S2) using specific antibodies. β-actin was used as a loading control. (C) The phosphorylation of endogenous CK2 substrates were analysed in lysates of HK-2 control (shCV) and silenced cells (shCK2α and shCK2β) by Western blot using Phospho-(Ser/Thr) CK2 Substrate (P-S/T3-100) antibody. Arrows indicate the bands that are more affected by <t>CK2β</t> down-regulation, whose quantification is shown in the bar graph. Tubulin was used as a loading control. A representative Western blot is shown in each case, while quantification (means ± SEM) is shown in the bar graphs. n = 4 for panel (A) and (B), n = 3 for panel (C).
Ck2β Floxed Mice, supplied by Inserm Transfert, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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ck2β subunits  (Gallus BioPharmaceuticals)
90
Gallus BioPharmaceuticals ck2β subunits
Alignment of PfCK2β1 and PfCK2β2 with Homo sapiens <t>CK2β.</t> ClustalW alignments of the following proteins were performed: Homo sapiens CK2β (HsCK2β; AAM50092); PfCK2β1 (AAN35637; PlasmoDB PF11_0048; 33% identical to HsCK2β), PfCK2β2 (CAD52554; PlasmoDB PF13_0232; 39% identical to HsCK2β). Note the long N-terminal extension and acidic insertion sequence in PfCK2β2. The sequence of shPfCK2β2 begins (after an artificially introduced initiating methionine) with residue E156, underlined. The cysteine residues thought to hold the zinc finger in place are indicated in bold text and with arrowheads. The potential autophosphorylation site on PfCK2β2 (SSEE) is indicated in bold. The export of CK2 as an ectokinase is mediated by CK2β residues 20 to 33 (44) (boxed, along with the equivalent residues in PfCK2β1 and PfCK2β2). The acidic region mentioned in the text is also boxed (D107 to E133 of PfCK2β2).
Ck2β Subunits, supplied by Gallus BioPharmaceuticals, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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ck2 β  (Biaffin Inc)
90
Biaffin Inc ck2 β
Alignment of PfCK2β1 and PfCK2β2 with Homo sapiens <t>CK2β.</t> ClustalW alignments of the following proteins were performed: Homo sapiens CK2β (HsCK2β; AAM50092); PfCK2β1 (AAN35637; PlasmoDB PF11_0048; 33% identical to HsCK2β), PfCK2β2 (CAD52554; PlasmoDB PF13_0232; 39% identical to HsCK2β). Note the long N-terminal extension and acidic insertion sequence in PfCK2β2. The sequence of shPfCK2β2 begins (after an artificially introduced initiating methionine) with residue E156, underlined. The cysteine residues thought to hold the zinc finger in place are indicated in bold text and with arrowheads. The potential autophosphorylation site on PfCK2β2 (SSEE) is indicated in bold. The export of CK2 as an ectokinase is mediated by CK2β residues 20 to 33 (44) (boxed, along with the equivalent residues in PfCK2β1 and PfCK2β2). The acidic region mentioned in the text is also boxed (D107 to E133 of PfCK2β2).
Ck2 β, supplied by Biaffin Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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ck2 β - by Bioz Stars, 2026-03
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ck2β active peptide  (Active Motif)
90
Active Motif ck2β active peptide
Alignment of PfCK2β1 and PfCK2β2 with Homo sapiens <t>CK2β.</t> ClustalW alignments of the following proteins were performed: Homo sapiens CK2β (HsCK2β; AAM50092); PfCK2β1 (AAN35637; PlasmoDB PF11_0048; 33% identical to HsCK2β), PfCK2β2 (CAD52554; PlasmoDB PF13_0232; 39% identical to HsCK2β). Note the long N-terminal extension and acidic insertion sequence in PfCK2β2. The sequence of shPfCK2β2 begins (after an artificially introduced initiating methionine) with residue E156, underlined. The cysteine residues thought to hold the zinc finger in place are indicated in bold text and with arrowheads. The potential autophosphorylation site on PfCK2β2 (SSEE) is indicated in bold. The export of CK2 as an ectokinase is mediated by CK2β residues 20 to 33 (44) (boxed, along with the equivalent residues in PfCK2β1 and PfCK2β2). The acidic region mentioned in the text is also boxed (D107 to E133 of PfCK2β2).
Ck2β Active Peptide, supplied by Active Motif, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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recombinant human ck2β  (Biaffin Inc)
90
Biaffin Inc recombinant human ck2β
Alignment of PfCK2β1 and PfCK2β2 with Homo sapiens <t>CK2β.</t> ClustalW alignments of the following proteins were performed: Homo sapiens CK2β (HsCK2β; AAM50092); PfCK2β1 (AAN35637; PlasmoDB PF11_0048; 33% identical to HsCK2β), PfCK2β2 (CAD52554; PlasmoDB PF13_0232; 39% identical to HsCK2β). Note the long N-terminal extension and acidic insertion sequence in PfCK2β2. The sequence of shPfCK2β2 begins (after an artificially introduced initiating methionine) with residue E156, underlined. The cysteine residues thought to hold the zinc finger in place are indicated in bold text and with arrowheads. The potential autophosphorylation site on PfCK2β2 (SSEE) is indicated in bold. The export of CK2 as an ectokinase is mediated by CK2β residues 20 to 33 (44) (boxed, along with the equivalent residues in PfCK2β1 and PfCK2β2). The acidic region mentioned in the text is also boxed (D107 to E133 of PfCK2β2).
Recombinant Human Ck2β, supplied by Biaffin Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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recombinant human ck2β - by Bioz Stars, 2026-03
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Image Search Results


a Immunoblotting showing the expression pattern of CK2β in mouse oocytes at different developmental stages. A total 150 oocytes were collected after being cultured for 0, 4, 8, and 12 h, corresponding to GV, GVBD, MI, and MII stages, respectively. β-actin was detected as an internal control. b Immunohistochemistry analysis of the expression pattern of CK2β in primordial, primary, secondary, and tertiary follicles at PD21. c Immunohistochemistry detection of CK2β loss in oocytes of Ck2β fl/fl ;GCre + mice. Scale bar: 50 μm

Journal: Cell Death & Disease

Article Title: Ablation of beta subunit of protein kinase CK2 in mouse oocytes causes follicle atresia and premature ovarian failure

doi: 10.1038/s41419-018-0505-1

Figure Lengend Snippet: a Immunoblotting showing the expression pattern of CK2β in mouse oocytes at different developmental stages. A total 150 oocytes were collected after being cultured for 0, 4, 8, and 12 h, corresponding to GV, GVBD, MI, and MII stages, respectively. β-actin was detected as an internal control. b Immunohistochemistry analysis of the expression pattern of CK2β in primordial, primary, secondary, and tertiary follicles at PD21. c Immunohistochemistry detection of CK2β loss in oocytes of Ck2β fl/fl ;GCre + mice. Scale bar: 50 μm

Article Snippet: Rabbit monoclonal anti-CK2β antibody (AJ1128b; ABGENT); mouse monoclonal anti-β-actin antibody (sc-47778; Santa Cruz); mouse monoclonal anti-MVH antibody (ab27591; abcam); rabbit monoclonal anti-p-Akt (S473) antibody (4046; Cell Signaling Technology, Inc.); rabbit monoclonal anti-p-Akt (T308) (13038; Cell Signaling Technology, Inc.); rabbit monoclonal anti-p-Akt (S129) (ab133458; abcam); rabbit monoclonal anti-AKT1/2/3 antibody (ab32505; abcam); rabbit monoclonal anti-PTEN antibody (9559; Cell Signaling Technology, Inc.); rabbit polyclonal anti-p-TSC2 (S1387) antibody (5584; Cell Signaling Technology, Inc.); rabbit monoclonal anti-TSC2 antibody (4308; Cell Signaling Technology, Inc.); rabbit monoclonal anti-p-mTOR (S2448) antibody (5536; Cell Signaling Technology, Inc.); rabbit monoclonal anti-p-S6K (T389) antibody (9234; Cell Signaling Technology, Inc.); rabbit monoclonal anti-p-rpS6 (S240/244) antibody (5364; Cell Signaling Technology, Inc.); rabbit polyclonal anti-GSK3α/β (S21/9) antibody (9331; Cell Signaling Technology, Inc.); rabbit monoclonal anti-GSK3α/β antibody (5676; Cell Signaling Technology, Inc.); rabbit polyclonal anti-p-FOXO1 (T24)/FOXO3a (T32) antibody (9464; Cell Signaling Technology, Inc.); rabbit monoclonal anti-FOXO1 antibody (2880; Cell Signaling Technology, Inc.); rabbit monoclonal anti-γH2AX antibody (9718; Cell Signaling Technology, Inc.); rabbit monoclonal anti-H2AX antibody (7631; Cell Signaling Technology, Inc.); rabbit polyclonal anti-p-CHK2 (T68) antibody (BS4043; Bioworld Technology, Inc.); rabbit polyclonal anti-CHK2 antibody (BS6791; Bioworld Technology, Inc.); rabbit monoclonal anti-p-p53 (S15) (12571; Cell Signaling Technology, Inc.); rabbit polyclonal anti-p53 antibody (BS3156; Bioworld Technology, Inc.); rabbit monoclonal anti-p63 (ab124762; abcam); mouse monoclonal anti-CK2α antibody (ab70774; abcam); rabbit polyclonal anti-CK2α’ antibody (BS6571; Bioworld Technology, Inc.); rabbit monoclonal anti-phospho-CK2 substrate [(pS/pT)DXE] (8738; Cell Signaling Technology, Inc.); secondary antibodies were purchased from Zhongshan Golden Bridge Biotechnology Co, Ltd (Beijing).

Techniques: Western Blot, Expressing, Cell Culture, Immunohistochemistry

a Comparison of the accumulative number of pups per Ck2β fl/fl female and Ck2β fl/fl ;GCre + female for 6 months. At least five mice of each genotype were used in this assay. b Ovary weight to body weight ratio of Ck2β fl/fl and Ck2β fl/fl ;GCre + mice at 3, 4, 6, and 8 weeks of age after birth. For each time point, at least three mice of each genotype were used for analysis. Data are presented as the mean ± SEM. P < 0.05(*), 0.01(**) or 0.001(***). c – j Representative ovarian histology of Ck2β fl/fl and Ck2β fl/fl ;GCre + mice of 3, 4, 6, and 8 weeks of age, respectively. Images c’ – j’ correspond to the partial magnification of images c – j . Yellow arrowheads in f’ , h’ , and j’ indicate atretic follicles. For each time point, at least three mice of each genotype were used for analysis. Scale bar: 100 μm

Journal: Cell Death & Disease

Article Title: Ablation of beta subunit of protein kinase CK2 in mouse oocytes causes follicle atresia and premature ovarian failure

doi: 10.1038/s41419-018-0505-1

Figure Lengend Snippet: a Comparison of the accumulative number of pups per Ck2β fl/fl female and Ck2β fl/fl ;GCre + female for 6 months. At least five mice of each genotype were used in this assay. b Ovary weight to body weight ratio of Ck2β fl/fl and Ck2β fl/fl ;GCre + mice at 3, 4, 6, and 8 weeks of age after birth. For each time point, at least three mice of each genotype were used for analysis. Data are presented as the mean ± SEM. P < 0.05(*), 0.01(**) or 0.001(***). c – j Representative ovarian histology of Ck2β fl/fl and Ck2β fl/fl ;GCre + mice of 3, 4, 6, and 8 weeks of age, respectively. Images c’ – j’ correspond to the partial magnification of images c – j . Yellow arrowheads in f’ , h’ , and j’ indicate atretic follicles. For each time point, at least three mice of each genotype were used for analysis. Scale bar: 100 μm

Article Snippet: Rabbit monoclonal anti-CK2β antibody (AJ1128b; ABGENT); mouse monoclonal anti-β-actin antibody (sc-47778; Santa Cruz); mouse monoclonal anti-MVH antibody (ab27591; abcam); rabbit monoclonal anti-p-Akt (S473) antibody (4046; Cell Signaling Technology, Inc.); rabbit monoclonal anti-p-Akt (T308) (13038; Cell Signaling Technology, Inc.); rabbit monoclonal anti-p-Akt (S129) (ab133458; abcam); rabbit monoclonal anti-AKT1/2/3 antibody (ab32505; abcam); rabbit monoclonal anti-PTEN antibody (9559; Cell Signaling Technology, Inc.); rabbit polyclonal anti-p-TSC2 (S1387) antibody (5584; Cell Signaling Technology, Inc.); rabbit monoclonal anti-TSC2 antibody (4308; Cell Signaling Technology, Inc.); rabbit monoclonal anti-p-mTOR (S2448) antibody (5536; Cell Signaling Technology, Inc.); rabbit monoclonal anti-p-S6K (T389) antibody (9234; Cell Signaling Technology, Inc.); rabbit monoclonal anti-p-rpS6 (S240/244) antibody (5364; Cell Signaling Technology, Inc.); rabbit polyclonal anti-GSK3α/β (S21/9) antibody (9331; Cell Signaling Technology, Inc.); rabbit monoclonal anti-GSK3α/β antibody (5676; Cell Signaling Technology, Inc.); rabbit polyclonal anti-p-FOXO1 (T24)/FOXO3a (T32) antibody (9464; Cell Signaling Technology, Inc.); rabbit monoclonal anti-FOXO1 antibody (2880; Cell Signaling Technology, Inc.); rabbit monoclonal anti-γH2AX antibody (9718; Cell Signaling Technology, Inc.); rabbit monoclonal anti-H2AX antibody (7631; Cell Signaling Technology, Inc.); rabbit polyclonal anti-p-CHK2 (T68) antibody (BS4043; Bioworld Technology, Inc.); rabbit polyclonal anti-CHK2 antibody (BS6791; Bioworld Technology, Inc.); rabbit monoclonal anti-p-p53 (S15) (12571; Cell Signaling Technology, Inc.); rabbit polyclonal anti-p53 antibody (BS3156; Bioworld Technology, Inc.); rabbit monoclonal anti-p63 (ab124762; abcam); mouse monoclonal anti-CK2α antibody (ab70774; abcam); rabbit polyclonal anti-CK2α’ antibody (BS6571; Bioworld Technology, Inc.); rabbit monoclonal anti-phospho-CK2 substrate [(pS/pT)DXE] (8738; Cell Signaling Technology, Inc.); secondary antibodies were purchased from Zhongshan Golden Bridge Biotechnology Co, Ltd (Beijing).

Techniques:

a Germ cells marker MVH immunohistochemistry of ovaries of Ck2β fl/fl and Ck2β fl/fl ;GCre + at 3 and 8 weeks after birth. At least three mice of each genotype were used in this assay. Scale bar: 100 μm. b TUNEL immunofluorescence staining of the ovaries of Ck2β fl/fl and Ck2β fl/fl ;GCre + at 4 weeks after birth. Green: TUNEL-positive signal; Blue: DAPI. At least three mice of each genotype were used for analysis. Scale bar: 100 μm

Journal: Cell Death & Disease

Article Title: Ablation of beta subunit of protein kinase CK2 in mouse oocytes causes follicle atresia and premature ovarian failure

doi: 10.1038/s41419-018-0505-1

Figure Lengend Snippet: a Germ cells marker MVH immunohistochemistry of ovaries of Ck2β fl/fl and Ck2β fl/fl ;GCre + at 3 and 8 weeks after birth. At least three mice of each genotype were used in this assay. Scale bar: 100 μm. b TUNEL immunofluorescence staining of the ovaries of Ck2β fl/fl and Ck2β fl/fl ;GCre + at 4 weeks after birth. Green: TUNEL-positive signal; Blue: DAPI. At least three mice of each genotype were used for analysis. Scale bar: 100 μm

Article Snippet: Rabbit monoclonal anti-CK2β antibody (AJ1128b; ABGENT); mouse monoclonal anti-β-actin antibody (sc-47778; Santa Cruz); mouse monoclonal anti-MVH antibody (ab27591; abcam); rabbit monoclonal anti-p-Akt (S473) antibody (4046; Cell Signaling Technology, Inc.); rabbit monoclonal anti-p-Akt (T308) (13038; Cell Signaling Technology, Inc.); rabbit monoclonal anti-p-Akt (S129) (ab133458; abcam); rabbit monoclonal anti-AKT1/2/3 antibody (ab32505; abcam); rabbit monoclonal anti-PTEN antibody (9559; Cell Signaling Technology, Inc.); rabbit polyclonal anti-p-TSC2 (S1387) antibody (5584; Cell Signaling Technology, Inc.); rabbit monoclonal anti-TSC2 antibody (4308; Cell Signaling Technology, Inc.); rabbit monoclonal anti-p-mTOR (S2448) antibody (5536; Cell Signaling Technology, Inc.); rabbit monoclonal anti-p-S6K (T389) antibody (9234; Cell Signaling Technology, Inc.); rabbit monoclonal anti-p-rpS6 (S240/244) antibody (5364; Cell Signaling Technology, Inc.); rabbit polyclonal anti-GSK3α/β (S21/9) antibody (9331; Cell Signaling Technology, Inc.); rabbit monoclonal anti-GSK3α/β antibody (5676; Cell Signaling Technology, Inc.); rabbit polyclonal anti-p-FOXO1 (T24)/FOXO3a (T32) antibody (9464; Cell Signaling Technology, Inc.); rabbit monoclonal anti-FOXO1 antibody (2880; Cell Signaling Technology, Inc.); rabbit monoclonal anti-γH2AX antibody (9718; Cell Signaling Technology, Inc.); rabbit monoclonal anti-H2AX antibody (7631; Cell Signaling Technology, Inc.); rabbit polyclonal anti-p-CHK2 (T68) antibody (BS4043; Bioworld Technology, Inc.); rabbit polyclonal anti-CHK2 antibody (BS6791; Bioworld Technology, Inc.); rabbit monoclonal anti-p-p53 (S15) (12571; Cell Signaling Technology, Inc.); rabbit polyclonal anti-p53 antibody (BS3156; Bioworld Technology, Inc.); rabbit monoclonal anti-p63 (ab124762; abcam); mouse monoclonal anti-CK2α antibody (ab70774; abcam); rabbit polyclonal anti-CK2α’ antibody (BS6571; Bioworld Technology, Inc.); rabbit monoclonal anti-phospho-CK2 substrate [(pS/pT)DXE] (8738; Cell Signaling Technology, Inc.); secondary antibodies were purchased from Zhongshan Golden Bridge Biotechnology Co, Ltd (Beijing).

Techniques: Marker, Immunohistochemistry, TUNEL Assay, Immunofluorescence, Staining

Immunoblotting detection of PI3K/AKT signaling in ovaries of Ck2β fl/fl and Ck2β fl/fl ;GCre + at 2 weeks after birth. The ovary lysates were collected at least from three mice of each genotype and immunoblotted for p-AKT (S473), p-AKT (T308), p-AKT (S129), AKT1/2/3, PTEN, p-TSC2 (S1387), TSC2, p-mTOR (S2448), p-S6K (T389), p-rpS6 (S240/244), p-GSKα/β (S21/9), GSKα/β, p-FOXO1 (T24)/FOXO3a (T32), FOXO1, and β-actin. Levels of β-action were used as internal control. Each experiment was repeated at least 2–3 times. Molecular mass is given in kilo Daltons

Journal: Cell Death & Disease

Article Title: Ablation of beta subunit of protein kinase CK2 in mouse oocytes causes follicle atresia and premature ovarian failure

doi: 10.1038/s41419-018-0505-1

Figure Lengend Snippet: Immunoblotting detection of PI3K/AKT signaling in ovaries of Ck2β fl/fl and Ck2β fl/fl ;GCre + at 2 weeks after birth. The ovary lysates were collected at least from three mice of each genotype and immunoblotted for p-AKT (S473), p-AKT (T308), p-AKT (S129), AKT1/2/3, PTEN, p-TSC2 (S1387), TSC2, p-mTOR (S2448), p-S6K (T389), p-rpS6 (S240/244), p-GSKα/β (S21/9), GSKα/β, p-FOXO1 (T24)/FOXO3a (T32), FOXO1, and β-actin. Levels of β-action were used as internal control. Each experiment was repeated at least 2–3 times. Molecular mass is given in kilo Daltons

Article Snippet: Rabbit monoclonal anti-CK2β antibody (AJ1128b; ABGENT); mouse monoclonal anti-β-actin antibody (sc-47778; Santa Cruz); mouse monoclonal anti-MVH antibody (ab27591; abcam); rabbit monoclonal anti-p-Akt (S473) antibody (4046; Cell Signaling Technology, Inc.); rabbit monoclonal anti-p-Akt (T308) (13038; Cell Signaling Technology, Inc.); rabbit monoclonal anti-p-Akt (S129) (ab133458; abcam); rabbit monoclonal anti-AKT1/2/3 antibody (ab32505; abcam); rabbit monoclonal anti-PTEN antibody (9559; Cell Signaling Technology, Inc.); rabbit polyclonal anti-p-TSC2 (S1387) antibody (5584; Cell Signaling Technology, Inc.); rabbit monoclonal anti-TSC2 antibody (4308; Cell Signaling Technology, Inc.); rabbit monoclonal anti-p-mTOR (S2448) antibody (5536; Cell Signaling Technology, Inc.); rabbit monoclonal anti-p-S6K (T389) antibody (9234; Cell Signaling Technology, Inc.); rabbit monoclonal anti-p-rpS6 (S240/244) antibody (5364; Cell Signaling Technology, Inc.); rabbit polyclonal anti-GSK3α/β (S21/9) antibody (9331; Cell Signaling Technology, Inc.); rabbit monoclonal anti-GSK3α/β antibody (5676; Cell Signaling Technology, Inc.); rabbit polyclonal anti-p-FOXO1 (T24)/FOXO3a (T32) antibody (9464; Cell Signaling Technology, Inc.); rabbit monoclonal anti-FOXO1 antibody (2880; Cell Signaling Technology, Inc.); rabbit monoclonal anti-γH2AX antibody (9718; Cell Signaling Technology, Inc.); rabbit monoclonal anti-H2AX antibody (7631; Cell Signaling Technology, Inc.); rabbit polyclonal anti-p-CHK2 (T68) antibody (BS4043; Bioworld Technology, Inc.); rabbit polyclonal anti-CHK2 antibody (BS6791; Bioworld Technology, Inc.); rabbit monoclonal anti-p-p53 (S15) (12571; Cell Signaling Technology, Inc.); rabbit polyclonal anti-p53 antibody (BS3156; Bioworld Technology, Inc.); rabbit monoclonal anti-p63 (ab124762; abcam); mouse monoclonal anti-CK2α antibody (ab70774; abcam); rabbit polyclonal anti-CK2α’ antibody (BS6571; Bioworld Technology, Inc.); rabbit monoclonal anti-phospho-CK2 substrate [(pS/pT)DXE] (8738; Cell Signaling Technology, Inc.); secondary antibodies were purchased from Zhongshan Golden Bridge Biotechnology Co, Ltd (Beijing).

Techniques: Western Blot

a Immunoblotting analysis of DNA damage response signaling in ovaries of Ck2β fl/fl and Ck2β fl/fl ;GCre + mice at 2 weeks after birth. The ovary lysates were collected at least from three mice of each genotype and immunoblotted for γH2AX, H2AX, p-CHK2 (T68), CHK2, p63, p-p53 (S15), p53 and β-actin. Level of β-actin was detected as internal control. Each experiment was repeated at least 2–3 times. Molecular mass is given in kilo Daltons. b Immunoblotting analysis of the expression of p53 and p63 in ovaries of Ck2β fl/fl and Ck2β fl/fl ;GCre + at 4 weeks after birth. The ovary lysates were obtained from at least three mice of each genotype and immunoblotted for p63, p-p53 (S15), p53 and β-actin. Level of β-actin was used as internal control. Each experiment was repeated at least three times. Molecular mass is given in kilo Daltons. c MVH and γH2AX immunofluorescent staining of 2-week-old ovarian sections from Ck2β fl/fl and Ck2β fl/fl ;GCre + mice. Green: γH2AX; Red: MVH; Blue: DAPI. At least three mice of each genotype were used in this assay. Scale bar: 50 μm

Journal: Cell Death & Disease

Article Title: Ablation of beta subunit of protein kinase CK2 in mouse oocytes causes follicle atresia and premature ovarian failure

doi: 10.1038/s41419-018-0505-1

Figure Lengend Snippet: a Immunoblotting analysis of DNA damage response signaling in ovaries of Ck2β fl/fl and Ck2β fl/fl ;GCre + mice at 2 weeks after birth. The ovary lysates were collected at least from three mice of each genotype and immunoblotted for γH2AX, H2AX, p-CHK2 (T68), CHK2, p63, p-p53 (S15), p53 and β-actin. Level of β-actin was detected as internal control. Each experiment was repeated at least 2–3 times. Molecular mass is given in kilo Daltons. b Immunoblotting analysis of the expression of p53 and p63 in ovaries of Ck2β fl/fl and Ck2β fl/fl ;GCre + at 4 weeks after birth. The ovary lysates were obtained from at least three mice of each genotype and immunoblotted for p63, p-p53 (S15), p53 and β-actin. Level of β-actin was used as internal control. Each experiment was repeated at least three times. Molecular mass is given in kilo Daltons. c MVH and γH2AX immunofluorescent staining of 2-week-old ovarian sections from Ck2β fl/fl and Ck2β fl/fl ;GCre + mice. Green: γH2AX; Red: MVH; Blue: DAPI. At least three mice of each genotype were used in this assay. Scale bar: 50 μm

Article Snippet: Rabbit monoclonal anti-CK2β antibody (AJ1128b; ABGENT); mouse monoclonal anti-β-actin antibody (sc-47778; Santa Cruz); mouse monoclonal anti-MVH antibody (ab27591; abcam); rabbit monoclonal anti-p-Akt (S473) antibody (4046; Cell Signaling Technology, Inc.); rabbit monoclonal anti-p-Akt (T308) (13038; Cell Signaling Technology, Inc.); rabbit monoclonal anti-p-Akt (S129) (ab133458; abcam); rabbit monoclonal anti-AKT1/2/3 antibody (ab32505; abcam); rabbit monoclonal anti-PTEN antibody (9559; Cell Signaling Technology, Inc.); rabbit polyclonal anti-p-TSC2 (S1387) antibody (5584; Cell Signaling Technology, Inc.); rabbit monoclonal anti-TSC2 antibody (4308; Cell Signaling Technology, Inc.); rabbit monoclonal anti-p-mTOR (S2448) antibody (5536; Cell Signaling Technology, Inc.); rabbit monoclonal anti-p-S6K (T389) antibody (9234; Cell Signaling Technology, Inc.); rabbit monoclonal anti-p-rpS6 (S240/244) antibody (5364; Cell Signaling Technology, Inc.); rabbit polyclonal anti-GSK3α/β (S21/9) antibody (9331; Cell Signaling Technology, Inc.); rabbit monoclonal anti-GSK3α/β antibody (5676; Cell Signaling Technology, Inc.); rabbit polyclonal anti-p-FOXO1 (T24)/FOXO3a (T32) antibody (9464; Cell Signaling Technology, Inc.); rabbit monoclonal anti-FOXO1 antibody (2880; Cell Signaling Technology, Inc.); rabbit monoclonal anti-γH2AX antibody (9718; Cell Signaling Technology, Inc.); rabbit monoclonal anti-H2AX antibody (7631; Cell Signaling Technology, Inc.); rabbit polyclonal anti-p-CHK2 (T68) antibody (BS4043; Bioworld Technology, Inc.); rabbit polyclonal anti-CHK2 antibody (BS6791; Bioworld Technology, Inc.); rabbit monoclonal anti-p-p53 (S15) (12571; Cell Signaling Technology, Inc.); rabbit polyclonal anti-p53 antibody (BS3156; Bioworld Technology, Inc.); rabbit monoclonal anti-p63 (ab124762; abcam); mouse monoclonal anti-CK2α antibody (ab70774; abcam); rabbit polyclonal anti-CK2α’ antibody (BS6571; Bioworld Technology, Inc.); rabbit monoclonal anti-phospho-CK2 substrate [(pS/pT)DXE] (8738; Cell Signaling Technology, Inc.); secondary antibodies were purchased from Zhongshan Golden Bridge Biotechnology Co, Ltd (Beijing).

Techniques: Western Blot, Expressing, Staining

a Immunoblotting detection of the expression of CK2α and CK2α’ in ovaries of Ck2β fl/fl and Ck2β fl/fl ;GCre + at 2 weeks after birth. b Immunoblotting detection of CK2 activity in ovaries of Ck2β fl/fl and Ck2β fl/fl ;GCre + at 2 weeks after birth. The obviously altered bands were marked with red arrows. The ovary lysates were obtained from at least three mice of each genotype and immunoblotted for CK2α, CK2α’, CK2β, and phospho-CK2 substrate, β-actin. Level of β-actin was used as internal control. Each experiment was repeated at least three times. Molecular mass is given in kilo Daltons

Journal: Cell Death & Disease

Article Title: Ablation of beta subunit of protein kinase CK2 in mouse oocytes causes follicle atresia and premature ovarian failure

doi: 10.1038/s41419-018-0505-1

Figure Lengend Snippet: a Immunoblotting detection of the expression of CK2α and CK2α’ in ovaries of Ck2β fl/fl and Ck2β fl/fl ;GCre + at 2 weeks after birth. b Immunoblotting detection of CK2 activity in ovaries of Ck2β fl/fl and Ck2β fl/fl ;GCre + at 2 weeks after birth. The obviously altered bands were marked with red arrows. The ovary lysates were obtained from at least three mice of each genotype and immunoblotted for CK2α, CK2α’, CK2β, and phospho-CK2 substrate, β-actin. Level of β-actin was used as internal control. Each experiment was repeated at least three times. Molecular mass is given in kilo Daltons

Article Snippet: Rabbit monoclonal anti-CK2β antibody (AJ1128b; ABGENT); mouse monoclonal anti-β-actin antibody (sc-47778; Santa Cruz); mouse monoclonal anti-MVH antibody (ab27591; abcam); rabbit monoclonal anti-p-Akt (S473) antibody (4046; Cell Signaling Technology, Inc.); rabbit monoclonal anti-p-Akt (T308) (13038; Cell Signaling Technology, Inc.); rabbit monoclonal anti-p-Akt (S129) (ab133458; abcam); rabbit monoclonal anti-AKT1/2/3 antibody (ab32505; abcam); rabbit monoclonal anti-PTEN antibody (9559; Cell Signaling Technology, Inc.); rabbit polyclonal anti-p-TSC2 (S1387) antibody (5584; Cell Signaling Technology, Inc.); rabbit monoclonal anti-TSC2 antibody (4308; Cell Signaling Technology, Inc.); rabbit monoclonal anti-p-mTOR (S2448) antibody (5536; Cell Signaling Technology, Inc.); rabbit monoclonal anti-p-S6K (T389) antibody (9234; Cell Signaling Technology, Inc.); rabbit monoclonal anti-p-rpS6 (S240/244) antibody (5364; Cell Signaling Technology, Inc.); rabbit polyclonal anti-GSK3α/β (S21/9) antibody (9331; Cell Signaling Technology, Inc.); rabbit monoclonal anti-GSK3α/β antibody (5676; Cell Signaling Technology, Inc.); rabbit polyclonal anti-p-FOXO1 (T24)/FOXO3a (T32) antibody (9464; Cell Signaling Technology, Inc.); rabbit monoclonal anti-FOXO1 antibody (2880; Cell Signaling Technology, Inc.); rabbit monoclonal anti-γH2AX antibody (9718; Cell Signaling Technology, Inc.); rabbit monoclonal anti-H2AX antibody (7631; Cell Signaling Technology, Inc.); rabbit polyclonal anti-p-CHK2 (T68) antibody (BS4043; Bioworld Technology, Inc.); rabbit polyclonal anti-CHK2 antibody (BS6791; Bioworld Technology, Inc.); rabbit monoclonal anti-p-p53 (S15) (12571; Cell Signaling Technology, Inc.); rabbit polyclonal anti-p53 antibody (BS3156; Bioworld Technology, Inc.); rabbit monoclonal anti-p63 (ab124762; abcam); mouse monoclonal anti-CK2α antibody (ab70774; abcam); rabbit polyclonal anti-CK2α’ antibody (BS6571; Bioworld Technology, Inc.); rabbit monoclonal anti-phospho-CK2 substrate [(pS/pT)DXE] (8738; Cell Signaling Technology, Inc.); secondary antibodies were purchased from Zhongshan Golden Bridge Biotechnology Co, Ltd (Beijing).

Techniques: Western Blot, Expressing, Activity Assay

CK2 interacts with NOXO1 in T84 colon epithelial cells under inflammatory conditions. ( A ) Immunoblots of NOXO1, CK2α/α′, CK2β, and β-actin in colon T84 cells stimulated with TNFα (5 ng/mL) or IL17 (50 ng/mL) individually or in combination for 24 hours at 37°C. Representative of 3 independent experiments. ( B ) ROS production was measured by chemiluminescence in T84 cells stimulated as in (A) . n = 3. ( C ) Immunoblots of CK2 α/α′ in NOXO1 IPP from T84 cells stimulated as in (A) . Representative of 4 independent experiments. ( D ) Densitometry analysis of CK2 α/α′ in NOXO1 IPP as in ( C ), normalized to CK2 α/α′expression in cell lysates; n = 4; mean ± SEM; one-way ANOVA with Tukey multiple comparisons test; ∗ P < .05. ( E ) Confocal microcopy of T84 cells co-stimulated or not with TNFα + IL17 for 24 hours at 37°C. NOXO1 ( green ), CK2 α/α′ ( red ), DAPI ( blue ). Scale bar: 10 μm. Representative of 6 independent experiments.

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Protein Kinase CK2 Acts as a Molecular Brake to Control NADPH Oxidase 1 Activation and Colon Inflammation

doi: 10.1016/j.jcmgh.2022.01.003

Figure Lengend Snippet: CK2 interacts with NOXO1 in T84 colon epithelial cells under inflammatory conditions. ( A ) Immunoblots of NOXO1, CK2α/α′, CK2β, and β-actin in colon T84 cells stimulated with TNFα (5 ng/mL) or IL17 (50 ng/mL) individually or in combination for 24 hours at 37°C. Representative of 3 independent experiments. ( B ) ROS production was measured by chemiluminescence in T84 cells stimulated as in (A) . n = 3. ( C ) Immunoblots of CK2 α/α′ in NOXO1 IPP from T84 cells stimulated as in (A) . Representative of 4 independent experiments. ( D ) Densitometry analysis of CK2 α/α′ in NOXO1 IPP as in ( C ), normalized to CK2 α/α′expression in cell lysates; n = 4; mean ± SEM; one-way ANOVA with Tukey multiple comparisons test; ∗ P < .05. ( E ) Confocal microcopy of T84 cells co-stimulated or not with TNFα + IL17 for 24 hours at 37°C. NOXO1 ( green ), CK2 α/α′ ( red ), DAPI ( blue ). Scale bar: 10 μm. Representative of 6 independent experiments.

Article Snippet: In some assays recombinant human CK2β protein (GeneTex Inc, Irvine, CA) was added.

Techniques: Western Blot, Expressing

CK2 interacts directly with NOXO1 through the N-terminal region mostly containing the PX domain. ( A ) Coomassie Blue staining of recombinant NOXO1, p47 PHOX , NOXA1, and p67 PHOX . ( B ) Dot-blot analysis of interaction between CK2 and recombinant NOXO1, p47 PHOX , NOXA1, or p67 PHOX . Representative of 3 independent experiments. ( C ) Schematic representation and Coomassie Blue staining of GST fusion proteins of the NOXO1 full-length (β isoform), N-terminal (1-157), and C-terminal regions (232-371). ( D ) Pull-down of CK2 from resting T84 cells lysates by full-length GST-NOXO1, GST-NOXO1 (1-157), GST-NOXO1 (232-371), or GST (control). Representative of 3 independent experiments. ( E ) Densitometry analysis of CK2 α/α′ pull-downed as in ( D ), normalized to GST-fusion proteins; n = 3; mean ± SEM; one-way ANOVA with Tukey multiple comparisons test; ∗∗∗∗ P < 0.0001.

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Protein Kinase CK2 Acts as a Molecular Brake to Control NADPH Oxidase 1 Activation and Colon Inflammation

doi: 10.1016/j.jcmgh.2022.01.003

Figure Lengend Snippet: CK2 interacts directly with NOXO1 through the N-terminal region mostly containing the PX domain. ( A ) Coomassie Blue staining of recombinant NOXO1, p47 PHOX , NOXA1, and p67 PHOX . ( B ) Dot-blot analysis of interaction between CK2 and recombinant NOXO1, p47 PHOX , NOXA1, or p67 PHOX . Representative of 3 independent experiments. ( C ) Schematic representation and Coomassie Blue staining of GST fusion proteins of the NOXO1 full-length (β isoform), N-terminal (1-157), and C-terminal regions (232-371). ( D ) Pull-down of CK2 from resting T84 cells lysates by full-length GST-NOXO1, GST-NOXO1 (1-157), GST-NOXO1 (232-371), or GST (control). Representative of 3 independent experiments. ( E ) Densitometry analysis of CK2 α/α′ pull-downed as in ( D ), normalized to GST-fusion proteins; n = 3; mean ± SEM; one-way ANOVA with Tukey multiple comparisons test; ∗∗∗∗ P < 0.0001.

Article Snippet: In some assays recombinant human CK2β protein (GeneTex Inc, Irvine, CA) was added.

Techniques: Staining, Recombinant, Dot Blot, Control

CK2 phosphorylates recombinant NOXO1 in vitro on several sites. ( A ) Autoradiography of recombinant NOXO1 phosphorylated with constitutively active CK2 in the presence of radioactive [γ -32P ] ATP. Time course ( left ), with or without CK2 ( right ). Representative of 3 independent experiments. ( B ) MS/MS protein coverage of recombinant NOXO1 after in vitro phosphorylation by CK2. Peptides were filtered according to 1% false discovery rate that corresponds to an identification score value >37.6 (–10lgP) with the software Peaks Studio Xpro. The confidence of modification sites is estimated by an Ascore, which calculates an ambiguity score as –10 × log 10 (p). The P value indicates the likelihood that the peptide is matched by chance (Ascore = 20 for P value of .01). Only confident modification sites with Ascore >20 were retained and their position labeled ( red ). ( C ) XIC of the heavily C-terminus phosphorylated peptide of m/z 753.997 showing 3 distinct elution peaks for 3 distinct phosphorylation sites. ( D ) Representative MS/MS spectrum of the phosphorylated C-terminus peptide at position 368. The confidence of modification sites is estimated by an Ascore (Ascore = 20 for P value of .01).

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Protein Kinase CK2 Acts as a Molecular Brake to Control NADPH Oxidase 1 Activation and Colon Inflammation

doi: 10.1016/j.jcmgh.2022.01.003

Figure Lengend Snippet: CK2 phosphorylates recombinant NOXO1 in vitro on several sites. ( A ) Autoradiography of recombinant NOXO1 phosphorylated with constitutively active CK2 in the presence of radioactive [γ -32P ] ATP. Time course ( left ), with or without CK2 ( right ). Representative of 3 independent experiments. ( B ) MS/MS protein coverage of recombinant NOXO1 after in vitro phosphorylation by CK2. Peptides were filtered according to 1% false discovery rate that corresponds to an identification score value >37.6 (–10lgP) with the software Peaks Studio Xpro. The confidence of modification sites is estimated by an Ascore, which calculates an ambiguity score as –10 × log 10 (p). The P value indicates the likelihood that the peptide is matched by chance (Ascore = 20 for P value of .01). Only confident modification sites with Ascore >20 were retained and their position labeled ( red ). ( C ) XIC of the heavily C-terminus phosphorylated peptide of m/z 753.997 showing 3 distinct elution peaks for 3 distinct phosphorylation sites. ( D ) Representative MS/MS spectrum of the phosphorylated C-terminus peptide at position 368. The confidence of modification sites is estimated by an Ascore (Ascore = 20 for P value of .01).

Article Snippet: In some assays recombinant human CK2β protein (GeneTex Inc, Irvine, CA) was added.

Techniques: Recombinant, In Vitro, Autoradiography, Tandem Mass Spectroscopy, Phospho-proteomics, Software, Modification, Labeling

Inhibition of CK2 enhances ROS production by NOX1 in colon T84 epithelial cells under inflammatory conditions. ( A ) ROS production was measured by chemiluminescence in T84 cells co-stimulated or not with TNFα + IL17 in presence or absence of 1 μmol/L CX-4945 after 24-hour incubation at 37°C. n = 3. ( B ) CK2 activity (top) assessed with the phospho-CK2-substrate [(pS/pT)DXE] antibody and NOXO1 expression (middle) in T84 cells co-stimulated as in (A) . Representative of 3 independent experiments. ( C ) Concentration-dependent effect of CX-4549 on ROS production by NOX1 in T84 cells co-stimulated as in (A) in presence or absence of various concentrations of CX-4945 for 24 hours at 37°C. Data were expressed as percentage of control (cells treated with TNFα + IL17 in absence of CX-4549); n = 7 per condition; mean ± SEM; one-way ANOVA with Tukey multiple comparisons test; ∗∗ P < .01. ( D ) Concentration-dependent effect of TBBz on ROS production by NOX1 measured by chemiluminescence in T84 cells co-stimulated as in (A) in the presence or absence of various concentrations of TBBz for 24 hours at 37°C. ( E ) Concentration-dependent effect of CX4945 on proliferation/cytotoxicity of T84 epithelial cells. n = 3 per condition; mean ± SEM; one-way ANOVA with Dunnett multiple comparisons test; ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001, ∗∗∗∗ P < .0001. ( F ) Concentration-dependent effect of TBBz on proliferation/cytotoxicity of T84 epithelial cells. n = 3 per condition; mean ± SEM.

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Protein Kinase CK2 Acts as a Molecular Brake to Control NADPH Oxidase 1 Activation and Colon Inflammation

doi: 10.1016/j.jcmgh.2022.01.003

Figure Lengend Snippet: Inhibition of CK2 enhances ROS production by NOX1 in colon T84 epithelial cells under inflammatory conditions. ( A ) ROS production was measured by chemiluminescence in T84 cells co-stimulated or not with TNFα + IL17 in presence or absence of 1 μmol/L CX-4945 after 24-hour incubation at 37°C. n = 3. ( B ) CK2 activity (top) assessed with the phospho-CK2-substrate [(pS/pT)DXE] antibody and NOXO1 expression (middle) in T84 cells co-stimulated as in (A) . Representative of 3 independent experiments. ( C ) Concentration-dependent effect of CX-4549 on ROS production by NOX1 in T84 cells co-stimulated as in (A) in presence or absence of various concentrations of CX-4945 for 24 hours at 37°C. Data were expressed as percentage of control (cells treated with TNFα + IL17 in absence of CX-4549); n = 7 per condition; mean ± SEM; one-way ANOVA with Tukey multiple comparisons test; ∗∗ P < .01. ( D ) Concentration-dependent effect of TBBz on ROS production by NOX1 measured by chemiluminescence in T84 cells co-stimulated as in (A) in the presence or absence of various concentrations of TBBz for 24 hours at 37°C. ( E ) Concentration-dependent effect of CX4945 on proliferation/cytotoxicity of T84 epithelial cells. n = 3 per condition; mean ± SEM; one-way ANOVA with Dunnett multiple comparisons test; ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001, ∗∗∗∗ P < .0001. ( F ) Concentration-dependent effect of TBBz on proliferation/cytotoxicity of T84 epithelial cells. n = 3 per condition; mean ± SEM.

Article Snippet: In some assays recombinant human CK2β protein (GeneTex Inc, Irvine, CA) was added.

Techniques: Inhibition, Incubation, Activity Assay, Expressing, Concentration Assay, Control

Inhibition of CK2 enhances ROS production by NOX1 in colon organoids under inflammatory conditions. ( A ) Representative immunoblots of NOXO1, CK2α/α′, CK2β, and β-actin in colon organoids stimulated as in <xref ref-type=Figure 1 A for 24 hours at 37°C. ( B ) Representative images of colon organoids established from colon biopsies of control patients. At day 8, organoids were co-stimulated or not with TNFα + IL17 in presence or absence of 1 μmol/L CX-4945 for 24 hours at 37°C. Scale bar: 100 μm. ( C ) ROS production was measured by chemiluminescence in colon organoids treated as in (B). ( D ) Data as in (C) were quantified and expressed as percentage of control (resting cells in the absence of CX-4549); n = 4 per condition; mean ± SEM; one-way ANOVA with Tukey multiple comparisons test; ∗ P < .05, ∗∗∗ P < .001. ( E ) ROS production by organoids treated as in ( B) was assessed by NBT reduction. Phase contrast microcopy with ×400 magnification. " width="100%" height="100%">

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Protein Kinase CK2 Acts as a Molecular Brake to Control NADPH Oxidase 1 Activation and Colon Inflammation

doi: 10.1016/j.jcmgh.2022.01.003

Figure Lengend Snippet: Inhibition of CK2 enhances ROS production by NOX1 in colon organoids under inflammatory conditions. ( A ) Representative immunoblots of NOXO1, CK2α/α′, CK2β, and β-actin in colon organoids stimulated as in Figure 1 A for 24 hours at 37°C. ( B ) Representative images of colon organoids established from colon biopsies of control patients. At day 8, organoids were co-stimulated or not with TNFα + IL17 in presence or absence of 1 μmol/L CX-4945 for 24 hours at 37°C. Scale bar: 100 μm. ( C ) ROS production was measured by chemiluminescence in colon organoids treated as in (B). ( D ) Data as in (C) were quantified and expressed as percentage of control (resting cells in the absence of CX-4549); n = 4 per condition; mean ± SEM; one-way ANOVA with Tukey multiple comparisons test; ∗ P < .05, ∗∗∗ P < .001. ( E ) ROS production by organoids treated as in ( B) was assessed by NBT reduction. Phase contrast microcopy with ×400 magnification.

Article Snippet: In some assays recombinant human CK2β protein (GeneTex Inc, Irvine, CA) was added.

Techniques: Inhibition, Western Blot, Control

CK2 activity is highly reduced and NOX1 expression is increased during TNBS-induced acute colitis. ( A ) Left, representative image of colons 24 hours after injection of TNBS or vehicle (CTL); middle, weight/length ratio changes after TNBS treatment; right, macroscopic lesions as assessed by Wallace score. Mean ± SEM, ≥10 animals; Mann-Whitney test; ∗∗∗∗ P < .0001. ( B ) Histologic images of colons 24 hours after TNBS-induced colitis (original magnification, ×200) as compared with CTL (vehicle). Scale bar: 50 μm. ( C ) CK2 activity in cytosolic and membrane fractions of colon homogenates assessed 24 hours after injection of TNBS or vehicle (CTL) with the phospho-CK2-substrate antibody. Four mice are shown (M1 to M4) for each group. ( D ) Densitometry analysis of p-CK2 substrates normalized to β-actin expression. Mean ± SEM from 10 animals; Kruskal-Wallis test with Dunn multiple comparisons test; ∗∗ P < .01, ∗∗∗ P < .001. ( E ) Immunoblots of CK2α/α′ and CK2β in colon homogenates 24 hours after injection of TNBS or vehicle (CTL). Four mice are shown (M1 to M4) for each group. ( F ) Densitometry analysis of CK2α/α′ and CK2β normalized to β-actin expression. Mean ± SEM, ≥6 animals; Mann-Whitney test; ∗∗ P < .01. ( G ) Immunoblots of NOX1 and p22 PHOX in colon homogenates 24 hours after injection of TNBS or vehicle (CTL). Two mice (M1 to M2) in the CTL group and 5 mice (M1 to M5) in the TNBS group are shown. ( H ) Densitometry analysis of NOX1 and p22 PHOX normalized to β-actin expression. Mean ± SEM, ≥8 animals; Mann-Whitney test; ∗∗ P < .01. ( I ) Expression of NOXO1 and NOXA1 RNA messengers during TNBS-induced acute colitis. Relative expression levels for each gene were calculated using the 2 -ΔΔCt method, with normalization to the average and GAPDH housekeeping genes. Mean ± SEM, 9 animals; Mann-Whitney test; ∗∗ P < .01.

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Protein Kinase CK2 Acts as a Molecular Brake to Control NADPH Oxidase 1 Activation and Colon Inflammation

doi: 10.1016/j.jcmgh.2022.01.003

Figure Lengend Snippet: CK2 activity is highly reduced and NOX1 expression is increased during TNBS-induced acute colitis. ( A ) Left, representative image of colons 24 hours after injection of TNBS or vehicle (CTL); middle, weight/length ratio changes after TNBS treatment; right, macroscopic lesions as assessed by Wallace score. Mean ± SEM, ≥10 animals; Mann-Whitney test; ∗∗∗∗ P < .0001. ( B ) Histologic images of colons 24 hours after TNBS-induced colitis (original magnification, ×200) as compared with CTL (vehicle). Scale bar: 50 μm. ( C ) CK2 activity in cytosolic and membrane fractions of colon homogenates assessed 24 hours after injection of TNBS or vehicle (CTL) with the phospho-CK2-substrate antibody. Four mice are shown (M1 to M4) for each group. ( D ) Densitometry analysis of p-CK2 substrates normalized to β-actin expression. Mean ± SEM from 10 animals; Kruskal-Wallis test with Dunn multiple comparisons test; ∗∗ P < .01, ∗∗∗ P < .001. ( E ) Immunoblots of CK2α/α′ and CK2β in colon homogenates 24 hours after injection of TNBS or vehicle (CTL). Four mice are shown (M1 to M4) for each group. ( F ) Densitometry analysis of CK2α/α′ and CK2β normalized to β-actin expression. Mean ± SEM, ≥6 animals; Mann-Whitney test; ∗∗ P < .01. ( G ) Immunoblots of NOX1 and p22 PHOX in colon homogenates 24 hours after injection of TNBS or vehicle (CTL). Two mice (M1 to M2) in the CTL group and 5 mice (M1 to M5) in the TNBS group are shown. ( H ) Densitometry analysis of NOX1 and p22 PHOX normalized to β-actin expression. Mean ± SEM, ≥8 animals; Mann-Whitney test; ∗∗ P < .01. ( I ) Expression of NOXO1 and NOXA1 RNA messengers during TNBS-induced acute colitis. Relative expression levels for each gene were calculated using the 2 -ΔΔCt method, with normalization to the average and GAPDH housekeeping genes. Mean ± SEM, 9 animals; Mann-Whitney test; ∗∗ P < .01.

Article Snippet: In some assays recombinant human CK2β protein (GeneTex Inc, Irvine, CA) was added.

Techniques: Activity Assay, Expressing, Injection, MANN-WHITNEY, Membrane, Western Blot

Effect of CK2β and CK2α overexpression on CK2 activity and ROS production in colon T84 epithelial cells. ( A ) Immunoblots of HA-CK2β and Myc-CK2α overexpressed individually in colon T84 cells. ( B ) CK2 activity assessed with the phospho-CK2-substrate [(pS/pT)DXE] antibody in T84 cells overexpressing CK2β or CK2α. ( C ) ROS production was measured by chemiluminescence in T84 cells overexpressing CK2β or CK2α. n = 3. ( D ) Effect of increasing concentrations of recombinant CK2β subunit on phosphorylation of NOXO1 by CK2 in vitro . Autoradiography (Autorad.) and Ponceau-Red are shown.

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Protein Kinase CK2 Acts as a Molecular Brake to Control NADPH Oxidase 1 Activation and Colon Inflammation

doi: 10.1016/j.jcmgh.2022.01.003

Figure Lengend Snippet: Effect of CK2β and CK2α overexpression on CK2 activity and ROS production in colon T84 epithelial cells. ( A ) Immunoblots of HA-CK2β and Myc-CK2α overexpressed individually in colon T84 cells. ( B ) CK2 activity assessed with the phospho-CK2-substrate [(pS/pT)DXE] antibody in T84 cells overexpressing CK2β or CK2α. ( C ) ROS production was measured by chemiluminescence in T84 cells overexpressing CK2β or CK2α. n = 3. ( D ) Effect of increasing concentrations of recombinant CK2β subunit on phosphorylation of NOXO1 by CK2 in vitro . Autoradiography (Autorad.) and Ponceau-Red are shown.

Article Snippet: In some assays recombinant human CK2β protein (GeneTex Inc, Irvine, CA) was added.

Techniques: Over Expression, Activity Assay, Western Blot, Recombinant, Phospho-proteomics, In Vitro, Autoradiography

The highly selective CK2 inhibitor CX-4945 exacerbates TNBS-induced colitis. ( A ) Representative images of colons 24 hours after injection of TNBS or vehicle (CTL) in mice treated with the CK2 inhibitor CX-4945 (25 mg/kg) or DMSO. ( B ) Histologic images of colons 24 hours after TNBS-induced colitis (original magnification, ×200) as compared with CTL (vehicle) in mice treated with the CK2 inhibitor CX-4945 (25 mg/kg) or DMSO. Scale bar: 50 μm. ( C ) Histologic damage of colon as assessed by the Ameho score in mice treated as in (B) . Mean ± SEM, ≥6 animals; Mann-Whitney test; ∗ P < .05. ( D ) Secretion of CXCL1 (pg. mg -1 protein) in colons of mice treated as in (B). Mean ± SEM from 4 animals (CTL groups with and without CX-4945) to 6 animals (TNBS groups with and without CX-4945), Mann-Whitney test; ∗ P < .05. ( E ) ROS production assessed ex vivo by NBT reduction in colon tissue of mice treated as in (B) . Formazan deposits were examined macroscopically. Representative of 4 mice in the CTL groups with and without CX-4945 and at least 7 mice in the TNBS groups with and without CX-4945. ( F ) MDA (nmol. mg -1 protein) content in the colon of mice treated as in (B) , normalized as compared with control. Mean ± SEM, ≥6 animals; Mann-Whitney test; ∗ P < .05.

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Protein Kinase CK2 Acts as a Molecular Brake to Control NADPH Oxidase 1 Activation and Colon Inflammation

doi: 10.1016/j.jcmgh.2022.01.003

Figure Lengend Snippet: The highly selective CK2 inhibitor CX-4945 exacerbates TNBS-induced colitis. ( A ) Representative images of colons 24 hours after injection of TNBS or vehicle (CTL) in mice treated with the CK2 inhibitor CX-4945 (25 mg/kg) or DMSO. ( B ) Histologic images of colons 24 hours after TNBS-induced colitis (original magnification, ×200) as compared with CTL (vehicle) in mice treated with the CK2 inhibitor CX-4945 (25 mg/kg) or DMSO. Scale bar: 50 μm. ( C ) Histologic damage of colon as assessed by the Ameho score in mice treated as in (B) . Mean ± SEM, ≥6 animals; Mann-Whitney test; ∗ P < .05. ( D ) Secretion of CXCL1 (pg. mg -1 protein) in colons of mice treated as in (B). Mean ± SEM from 4 animals (CTL groups with and without CX-4945) to 6 animals (TNBS groups with and without CX-4945), Mann-Whitney test; ∗ P < .05. ( E ) ROS production assessed ex vivo by NBT reduction in colon tissue of mice treated as in (B) . Formazan deposits were examined macroscopically. Representative of 4 mice in the CTL groups with and without CX-4945 and at least 7 mice in the TNBS groups with and without CX-4945. ( F ) MDA (nmol. mg -1 protein) content in the colon of mice treated as in (B) , normalized as compared with control. Mean ± SEM, ≥6 animals; Mann-Whitney test; ∗ P < .05.

Article Snippet: In some assays recombinant human CK2β protein (GeneTex Inc, Irvine, CA) was added.

Techniques: Injection, MANN-WHITNEY, Ex Vivo, Control

Expression of NOX1 and p22 PHOX during TNBS-induced acute colitis in the presence or absence of CX-4945. Immunoblots of NOX1 and p22 PHOX in colon homogenates 24 hours after injection of TNBS or vehicle (CTL) in mice treated with the CK2 inhibitor CX-4945 (25 mg/kg) or DMSO. Results from 2 mice (M1 to M2) in the CTL group and 4 mice (M1 to M4) in the TNBS group are shown.

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Protein Kinase CK2 Acts as a Molecular Brake to Control NADPH Oxidase 1 Activation and Colon Inflammation

doi: 10.1016/j.jcmgh.2022.01.003

Figure Lengend Snippet: Expression of NOX1 and p22 PHOX during TNBS-induced acute colitis in the presence or absence of CX-4945. Immunoblots of NOX1 and p22 PHOX in colon homogenates 24 hours after injection of TNBS or vehicle (CTL) in mice treated with the CK2 inhibitor CX-4945 (25 mg/kg) or DMSO. Results from 2 mice (M1 to M2) in the CTL group and 4 mice (M1 to M4) in the TNBS group are shown.

Article Snippet: In some assays recombinant human CK2β protein (GeneTex Inc, Irvine, CA) was added.

Techniques: Expressing, Western Blot, Injection

(A) HK-2 control (shCV) and silenced cells (shCK2α and shCK2β) were analysed for the expression of each CK2 subunit (α,α’ or β) and for the phosphorylation state of the CK2 target Akt S129. The 1AD9 anti-CK2α antibody detects also CK2α’; the arrows indicate the migration of each catalytic isoform. (B) Detection of total and phosphorylated eIF2β(S2) using specific antibodies. β-actin was used as a loading control. (C) The phosphorylation of endogenous CK2 substrates were analysed in lysates of HK-2 control (shCV) and silenced cells (shCK2α and shCK2β) by Western blot using Phospho-(Ser/Thr) CK2 Substrate (P-S/T3-100) antibody. Arrows indicate the bands that are more affected by CK2β down-regulation, whose quantification is shown in the bar graph. Tubulin was used as a loading control. A representative Western blot is shown in each case, while quantification (means ± SEM) is shown in the bar graphs. n = 4 for panel (A) and (B), n = 3 for panel (C).

Journal: PLoS ONE

Article Title: Effects of CK2β subunit down-regulation on Akt signalling in HK-2 renal cells

doi: 10.1371/journal.pone.0227340

Figure Lengend Snippet: (A) HK-2 control (shCV) and silenced cells (shCK2α and shCK2β) were analysed for the expression of each CK2 subunit (α,α’ or β) and for the phosphorylation state of the CK2 target Akt S129. The 1AD9 anti-CK2α antibody detects also CK2α’; the arrows indicate the migration of each catalytic isoform. (B) Detection of total and phosphorylated eIF2β(S2) using specific antibodies. β-actin was used as a loading control. (C) The phosphorylation of endogenous CK2 substrates were analysed in lysates of HK-2 control (shCV) and silenced cells (shCK2α and shCK2β) by Western blot using Phospho-(Ser/Thr) CK2 Substrate (P-S/T3-100) antibody. Arrows indicate the bands that are more affected by CK2β down-regulation, whose quantification is shown in the bar graph. Tubulin was used as a loading control. A representative Western blot is shown in each case, while quantification (means ± SEM) is shown in the bar graphs. n = 4 for panel (A) and (B), n = 3 for panel (C).

Article Snippet: For the CK2β siRNA silencing experiments, HK-2 and HEK293T cells were transfected with 30 nM CK2β specific siRNA (sequence target: 5'-GCCATGGTGAAGCTCTACT-3' ) or scrambled siRNA using the GenMute siRNA Transfection Reagent (SignaGen Laboratories, Rockville, MD, USA) according to the manufacturer’s recommendations.

Techniques: Expressing, Migration, Western Blot

(A) Increasing amounts of CK2α (1.25, 2.5 and 5 ng), CK2α’ (40, 80 and 160 ng) or CK2α2β2 (2.5, 5 and 10 ng), corresponding to equal activity towards CK2-tide peptide, were incubated with 0.3 μg of active Akt1 or 0.3 μg of inactive Akt1, or 1 μg β-casein as a control. In the right panel, 0.3 μg of active form of Akt was incubated with CK2α (1.25 ng), CK2α2β2 (5 ng), CK2α’ (40 ng) or CK2α’2β2 (25 ng) in the presence of 150 mM NaCl. The amount of each enzyme used was equally active against the CK2 specific peptide (CK2-tide: RRRADDSDDDDD ) as determined by kinase activity assays (not shown). Upon radioactive phosphorylation, proteins were separated by SDS-PAGE. A representative digital autoradiography of the dried gel is shown. The bar graph on the right shows the relative quantitation of the bands (means ± SEM, n = 3), performed by analysis with CyclonePlus Storage Phosphor System, (PerkinElmer); the unit amounts of each isoform are indicated, and activity is reported as % of that measured with 5 units of CK2α. (B) shCV HK-2 and shCK2β HK-2 were transiently transfected with CK2β pCMV-HA vector. Representative Western blot analysis with the indicated antibodies is shown. β-actin was used as a loading control. Quantification (p-Akt(S129)/ AKT) is shown on the bar graph on the right (means ± SEM, n = 2).

Journal: PLoS ONE

Article Title: Effects of CK2β subunit down-regulation on Akt signalling in HK-2 renal cells

doi: 10.1371/journal.pone.0227340

Figure Lengend Snippet: (A) Increasing amounts of CK2α (1.25, 2.5 and 5 ng), CK2α’ (40, 80 and 160 ng) or CK2α2β2 (2.5, 5 and 10 ng), corresponding to equal activity towards CK2-tide peptide, were incubated with 0.3 μg of active Akt1 or 0.3 μg of inactive Akt1, or 1 μg β-casein as a control. In the right panel, 0.3 μg of active form of Akt was incubated with CK2α (1.25 ng), CK2α2β2 (5 ng), CK2α’ (40 ng) or CK2α’2β2 (25 ng) in the presence of 150 mM NaCl. The amount of each enzyme used was equally active against the CK2 specific peptide (CK2-tide: RRRADDSDDDDD ) as determined by kinase activity assays (not shown). Upon radioactive phosphorylation, proteins were separated by SDS-PAGE. A representative digital autoradiography of the dried gel is shown. The bar graph on the right shows the relative quantitation of the bands (means ± SEM, n = 3), performed by analysis with CyclonePlus Storage Phosphor System, (PerkinElmer); the unit amounts of each isoform are indicated, and activity is reported as % of that measured with 5 units of CK2α. (B) shCV HK-2 and shCK2β HK-2 were transiently transfected with CK2β pCMV-HA vector. Representative Western blot analysis with the indicated antibodies is shown. β-actin was used as a loading control. Quantification (p-Akt(S129)/ AKT) is shown on the bar graph on the right (means ± SEM, n = 2).

Article Snippet: For the CK2β siRNA silencing experiments, HK-2 and HEK293T cells were transfected with 30 nM CK2β specific siRNA (sequence target: 5'-GCCATGGTGAAGCTCTACT-3' ) or scrambled siRNA using the GenMute siRNA Transfection Reagent (SignaGen Laboratories, Rockville, MD, USA) according to the manufacturer’s recommendations.

Techniques: Activity Assay, Incubation, SDS Page, Autoradiography, Quantitation Assay, Transfection, Plasmid Preparation, Western Blot

The components whose specific phosphorylation has been checked in this work are indicated in grey. The effects of CK2β silencing (A) or CK2 inhibition by CX-4945 (B) in HK-2 cells are highlighted by colours (upregulation in green, downregulation in red, unchanged in blue). The effect of CX-4945 on Akt pS129 is inferred from previous work of our and other laboratories (e.g. [ , ]).

Journal: PLoS ONE

Article Title: Effects of CK2β subunit down-regulation on Akt signalling in HK-2 renal cells

doi: 10.1371/journal.pone.0227340

Figure Lengend Snippet: The components whose specific phosphorylation has been checked in this work are indicated in grey. The effects of CK2β silencing (A) or CK2 inhibition by CX-4945 (B) in HK-2 cells are highlighted by colours (upregulation in green, downregulation in red, unchanged in blue). The effect of CX-4945 on Akt pS129 is inferred from previous work of our and other laboratories (e.g. [ , ]).

Article Snippet: For the CK2β siRNA silencing experiments, HK-2 and HEK293T cells were transfected with 30 nM CK2β specific siRNA (sequence target: 5'-GCCATGGTGAAGCTCTACT-3' ) or scrambled siRNA using the GenMute siRNA Transfection Reagent (SignaGen Laboratories, Rockville, MD, USA) according to the manufacturer’s recommendations.

Techniques: Inhibition

Alignment of PfCK2β1 and PfCK2β2 with Homo sapiens CK2β. ClustalW alignments of the following proteins were performed: Homo sapiens CK2β (HsCK2β; AAM50092); PfCK2β1 (AAN35637; PlasmoDB PF11_0048; 33% identical to HsCK2β), PfCK2β2 (CAD52554; PlasmoDB PF13_0232; 39% identical to HsCK2β). Note the long N-terminal extension and acidic insertion sequence in PfCK2β2. The sequence of shPfCK2β2 begins (after an artificially introduced initiating methionine) with residue E156, underlined. The cysteine residues thought to hold the zinc finger in place are indicated in bold text and with arrowheads. The potential autophosphorylation site on PfCK2β2 (SSEE) is indicated in bold. The export of CK2 as an ectokinase is mediated by CK2β residues 20 to 33 (44) (boxed, along with the equivalent residues in PfCK2β1 and PfCK2β2). The acidic region mentioned in the text is also boxed (D107 to E133 of PfCK2β2).

Journal:

Article Title: Functional Analysis of Protein Kinase CK2 of the Human Malaria Parasite Plasmodium falciparum ▿ †

doi: 10.1128/EC.00334-08

Figure Lengend Snippet: Alignment of PfCK2β1 and PfCK2β2 with Homo sapiens CK2β. ClustalW alignments of the following proteins were performed: Homo sapiens CK2β (HsCK2β; AAM50092); PfCK2β1 (AAN35637; PlasmoDB PF11_0048; 33% identical to HsCK2β), PfCK2β2 (CAD52554; PlasmoDB PF13_0232; 39% identical to HsCK2β). Note the long N-terminal extension and acidic insertion sequence in PfCK2β2. The sequence of shPfCK2β2 begins (after an artificially introduced initiating methionine) with residue E156, underlined. The cysteine residues thought to hold the zinc finger in place are indicated in bold text and with arrowheads. The potential autophosphorylation site on PfCK2β2 (SSEE) is indicated in bold. The export of CK2 as an ectokinase is mediated by CK2β residues 20 to 33 (44) (boxed, along with the equivalent residues in PfCK2β1 and PfCK2β2). The acidic region mentioned in the text is also boxed (D107 to E133 of PfCK2β2).

Article Snippet: Most CK2β subunits from vertebrates have only eight amino acids prior to this conserved residue ( Homo sapiens, Gallus gallus, Mus musculus, Xenopus tropicalis, Bos taurus , and Danio rerio ); this N-terminal extension is expanded in yeast ( Saccharomyces cerevisiae , 37 residues), trypanosomatids ( Trypanosoma brucei , 27 residues; Leishmania major , 21 residues), plants ( Arabidopsis thaliana , 100 residues; Oryza sativa , 92 residues), and alveolates ( Cryptosporidium parvum , 27 residues; Theileria parva , 34 residues).

Techniques: Sequencing, Residue

PfCK2α can be distinguished from HsCK2α by small molecule inhibitors. Two small molecules, ML-7 and Rottlerin, were identified in a primary screen as inhibiting the activity of PfCK2α to below 10% of the uninhibited enzyme. The inhibitors Rottlerin and ML-7 were included in increasing concentrations in kinase assays with 25 μM ATP, 36 ng of enzyme, and the peptide RRREDEESDDEE as substrate. Activity was measured using the phosphocellulose assay method, and results were scored as a percentage of the control (no inhibitor). (A) ML-7. (B) Rottlerin. Mean values from two experiments are shown, with the error bars representing the standard deviations. (C) The classical CK2 inhibitor, TBB, has a similar inhibitory profile for PfCK2α and HsCK2α.

Journal:

Article Title: Functional Analysis of Protein Kinase CK2 of the Human Malaria Parasite Plasmodium falciparum ▿ †

doi: 10.1128/EC.00334-08

Figure Lengend Snippet: PfCK2α can be distinguished from HsCK2α by small molecule inhibitors. Two small molecules, ML-7 and Rottlerin, were identified in a primary screen as inhibiting the activity of PfCK2α to below 10% of the uninhibited enzyme. The inhibitors Rottlerin and ML-7 were included in increasing concentrations in kinase assays with 25 μM ATP, 36 ng of enzyme, and the peptide RRREDEESDDEE as substrate. Activity was measured using the phosphocellulose assay method, and results were scored as a percentage of the control (no inhibitor). (A) ML-7. (B) Rottlerin. Mean values from two experiments are shown, with the error bars representing the standard deviations. (C) The classical CK2 inhibitor, TBB, has a similar inhibitory profile for PfCK2α and HsCK2α.

Article Snippet: Most CK2β subunits from vertebrates have only eight amino acids prior to this conserved residue ( Homo sapiens, Gallus gallus, Mus musculus, Xenopus tropicalis, Bos taurus , and Danio rerio ); this N-terminal extension is expanded in yeast ( Saccharomyces cerevisiae , 37 residues), trypanosomatids ( Trypanosoma brucei , 27 residues; Leishmania major , 21 residues), plants ( Arabidopsis thaliana , 100 residues; Oryza sativa , 92 residues), and alveolates ( Cryptosporidium parvum , 27 residues; Theileria parva , 34 residues).

Techniques: Activity Assay, Control

PfCK2α and HsCK2β interact in vitro. To further test the interactions of the alpha and beta subunits, two substrates that are phosphorylated by the CK2 holoenzyme and not by the CK2α subunit alone were included in kinase assays with mixtures of human and P. falciparum alpha and beta subunits. (A) Phosphorylation of the eIF2β[1-22] peptide (40, 45) by PfCK2α-His or HsCK2α in the presence and absence of GST-PfCK2β1, GST-shPfCK2β2, or HsCK2β was measured by kinase assays, and the amount of radiolabel incorporated into the peptide was counted by scintillation. Results are shown as the means of two experiments, with the error bars representing the standard deviations. (B) Phosphorylation of the GST-Olig2[1-177] protein (27) by PfCK2α-His (lanes 1 to 4) or HsCK2α (lanes 5 to 8) alone (lanes 1 and 5) or in the presence of GST-PfCK2β1 (lanes 2 and 6), GST-shPfCK2β2 (lanes 3 and 7), or HsCK2β (lanes 4 and 8). Top, autoradiogram; bottom, corresponding Coomassie blue-stained gel of the kinase assay.

Journal:

Article Title: Functional Analysis of Protein Kinase CK2 of the Human Malaria Parasite Plasmodium falciparum ▿ †

doi: 10.1128/EC.00334-08

Figure Lengend Snippet: PfCK2α and HsCK2β interact in vitro. To further test the interactions of the alpha and beta subunits, two substrates that are phosphorylated by the CK2 holoenzyme and not by the CK2α subunit alone were included in kinase assays with mixtures of human and P. falciparum alpha and beta subunits. (A) Phosphorylation of the eIF2β[1-22] peptide (40, 45) by PfCK2α-His or HsCK2α in the presence and absence of GST-PfCK2β1, GST-shPfCK2β2, or HsCK2β was measured by kinase assays, and the amount of radiolabel incorporated into the peptide was counted by scintillation. Results are shown as the means of two experiments, with the error bars representing the standard deviations. (B) Phosphorylation of the GST-Olig2[1-177] protein (27) by PfCK2α-His (lanes 1 to 4) or HsCK2α (lanes 5 to 8) alone (lanes 1 and 5) or in the presence of GST-PfCK2β1 (lanes 2 and 6), GST-shPfCK2β2 (lanes 3 and 7), or HsCK2β (lanes 4 and 8). Top, autoradiogram; bottom, corresponding Coomassie blue-stained gel of the kinase assay.

Article Snippet: Most CK2β subunits from vertebrates have only eight amino acids prior to this conserved residue ( Homo sapiens, Gallus gallus, Mus musculus, Xenopus tropicalis, Bos taurus , and Danio rerio ); this N-terminal extension is expanded in yeast ( Saccharomyces cerevisiae , 37 residues), trypanosomatids ( Trypanosoma brucei , 27 residues; Leishmania major , 21 residues), plants ( Arabidopsis thaliana , 100 residues; Oryza sativa , 92 residues), and alveolates ( Cryptosporidium parvum , 27 residues; Theileria parva , 34 residues).

Techniques: In Vitro, Phospho-proteomics, Staining, Kinase Assay