Journal: Science signaling

Article Title: A post-translational modification code for CFTR maturation is altered in cystic fibrosis

doi: 10.1126/scisignal.aan7984

Figure Lengend Snippet: A. CX-4945 mediated inhibition of CK2α prevents phosphorylation of wild-type CFTR at Thr421, Ser422 and Ser427 as quantified with spiked-in synthetic, heavy isotope-labeled peptides. Data are means ± S.D. of n=3 experiments; statistical analysis noted below. B. Experimental outline for AHA labeling and enrichment of newly synthesized CFTR by click chemistry. C. Western blot of input lysate for CFTR-IP before click reaction. D. Western blot showing that wild-type CFTR maturation is dependent on CK2α phosphorylation in the RI element. Blots are representative of n=3 experiments. E. Quantification of newly synthesized immature (band B) and mature (band C) CFTR relative to time point 0 (h, hours). F. Western blot of mature CFTR (band C) in FRT cells expressing various CFTR mutants. G. Phosphorylation at Thr421, Ser422 and Ser427 in the various CFTR mutants described in (F) was quantified with spiked-in synthetic heavy isotope-labeled peptides, relative to the amount of mature CFTR present. Data are mean ± S.D. of ≥ n=4 independent measurements per site; statistical analysis noted below. (H to K) AHA pulse-chase assays in primary human bronchial epithelial (NHBE) cells (H) and FRT cells expressing G551D or wild-type CFTR (K) treated with CX-4945 or DMSO. Subsequent quantification of newly synthesized immature CFTR (band B) and mature CFTR (band C) at the time points indicated relative to time 0 (I and K). Top panels show input and loading control. SA-HRP, streptavidin-conjugated horse radish peroxidase. Time of AHA labeling before CFTR immunoprecipitation is indicated. Blots are representative, and the data are means ± S.D. of n=3 independent biological replicates. *P < 0.0332, **P < 0.0021, ***P < 0.0002, and ****P < 0.0001 by unpaired, two-tailed t- tests with Bonferroni-Sidak correction (A) or one-Way ANOVA with Tukey post-test (I and K).

Article Snippet: Treatment with 5 – 10 μM VX809 (Selleck, Houston, TX), 10 μM CX-4945 (Santa Cruz Biotechnologies, Dallas, TX) or vehicle (DMSO), was carried out for 20 hours or as indicated.

Techniques: Inhibition, Labeling, Synthesized, Western Blot, Expressing, Pulse Chase, Immunoprecipitation, Two Tailed Test

Journal: Science signaling

Article Title: A post-translational modification code for CFTR maturation is altered in cystic fibrosis

doi: 10.1126/scisignal.aan7984

Figure Lengend Snippet: Degradation network identified for wild-type CFTR, wild-type CFTR treated with CX-4945, or ΔF508 CFTR in 16HBE41o- and CFBE41o- cells shows recruitment of ΔF508- specific interactions in wild-type CFTR expressing cells treated with CX-4945 (red nodes). Data represent independent biological replicates (wt n=8, ΔF508 n=8, wt+CX-4945 n=3).

Article Snippet: Treatment with 5 – 10 μM VX809 (Selleck, Houston, TX), 10 μM CX-4945 (Santa Cruz Biotechnologies, Dallas, TX) or vehicle (DMSO), was carried out for 20 hours or as indicated.

Techniques: Expressing

Journal: Science signaling

Article Title: A post-translational modification code for CFTR maturation is altered in cystic fibrosis

doi: 10.1126/scisignal.aan7984

Figure Lengend Snippet: A. Quantification of phosphorylated sites Ser422 and Ser427, methylation site Lys442 and ubiquitination site Lys420 at 37°C (red bars) or at permissive temperature of 30°C (blue bars). B. Quantification of ΔF508 CFTR phosphorylation at Thr421, Ser422 and Ser427 by synthetic heavy-labeled peptides shows reduced phosphorylation upon shRNA- mediated knockdown of CSNK2A1. C. Western blot showing that shRNA-mediated knockdown of CSNK2A1 impairs ΔF508 CFTR maturation at permissive temperature. Detection of b-actin was used as loading control. D. Representative traces of forskolin (F) and genistein (G) stimulated ΔF508 CFTR short circuit currents (/sc) showing that knockdown of CSNK2A1 (right) or treatment with 10 μM CX-4945 (left) prevent temperature shift (28°C) induced rescue of ΔF508 CFTR channel activity. Specificity of the current is established by treatment with CFTR inhibitor 172 (I). E. Quantification of Δ/sc currents as log2 fold change relative to control cells. All data are representative or mean ± S.D. of n=3 independent biological replicates. *P < 0.0332, **P < 0.0021, ***P < 0.0002, and ****P < 0.0001 by one-Way ANOVA with Bonferroni post-test.

Article Snippet: Treatment with 5 – 10 μM VX809 (Selleck, Houston, TX), 10 μM CX-4945 (Santa Cruz Biotechnologies, Dallas, TX) or vehicle (DMSO), was carried out for 20 hours or as indicated.

Techniques: Methylation, Labeling, shRNA, Western Blot, Activity Assay

Journal: bioRxiv

Article Title: CK2 inhibitor, CX-4945, enhances BH3 priming and promotes apoptosis of venetoclax-resistant AML by targeting antiapoptotic proteins

doi: 10.64898/2025.12.24.696284

Figure Lengend Snippet: A) AML cell lines including both VEN-susceptible and resistant (Molm-13, Molm-13/VR, HL-60, HL-60/VR, U937), and AML PDX cells (2016-1, 2016-7) were treated with different concentrations of CX-4945 and VEN alone or in combination with indicated doses for 48 h. Cell viability was assessed using WST reagent and cell viability was calculated relative to vehicle-treated cells as 100%. B) ZIP synergy scores for CX-4945 and VEN combination in different AML cell lines and PDX cells were shown. ZIP scores greater than 10 indicates ‘synergy’, and 0-10 indicate ‘additive’ activity between CX-4945 and VEN. C-D) AML cells (Molm-13, Molm-13/VR, PDX 2016-7) were treated with CX-4945 and VEN alone or in combination for 24 h and stained with Annexin V/7AAD for analyzing apoptosis by flow cytometry ( C ). Relative apoptosis was calculated by normalizing to vehicle treated cells ( D ). The data are presented as mean ± SD (n=2-4). *p<0.05, **p<0.01, ***p<0.001, and ****p<0.0001 by one-way ANOVA (Holm-Sidak’s multiple comparisons test) denotes statistical significance. E) AML cell lines (Molm-13, Molm-13/VR) were treated with CX-4945 and VEN alone or in combination for 24 h and the surface expression of chemoresistance markers (CD47 and CD123) was analyzed by flow cytometry. F) Schematic showing the workflow for dynamic BH3 profiling to measure drug-induced priming of BH3 peptides in AML cells. The difference in Cyt c release (%) with and without drug treatment was presented as delta priming. Created with BioRender.com . G) Molm-13 and Molm-13/VR cells were tested for Cyt C release by priming with BH3 peptides with and without CX-4945 treatment and calculated the delta priming. The data are presented as mean ± SEM (n=2) and analyzed by two-way ANOVA (Sidak’s multiple comparisons test).

Article Snippet: CX-4945 (#HY-50855B) and venetoclax (#HY-15531) were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Activity Assay, Staining, Flow Cytometry, Expressing

Journal: bioRxiv

Article Title: CK2 inhibitor, CX-4945, enhances BH3 priming and promotes apoptosis of venetoclax-resistant AML by targeting antiapoptotic proteins

doi: 10.64898/2025.12.24.696284

Figure Lengend Snippet: A) The common mechanisms that govern cellular MCL-1 levels at transcription, translation, and post-translation levels are depicted. The signaling cascade mediated through CK2, PI3K/AKT, and mTOR regulate transcription and translation of MCL-1 isoforms (L-large, S-short, ES-extra short). MCL-1L (anti-apoptotic) undergoes caspase-mediated cleavage to form pro-apoptotic shorter MCL-1 isoforms (S & ES) that can induce cellular apoptosis independent of BAX and BAK. Created using BioRender.com . B) The levels of MCL-1 isoforms (L and ES) were measured by immunoblotting in AML cell lines and PDX cells after 24h of treatment with CX-4945 and VEN alone or in combination. C) Quantification of MCL-1 transcript levels after treatment with CX-4945 and VEN alone or in combination for 24h in indicated AML cell lines and PDX cells by qRT-PCR. The data are presented as mean ± SD (n=3) and analyzed by two-way ANOVA (Tukey’s multiple comparisons test). *p<0.05, **p<0.01, ***p<0.001, and ****p<0.0001 are considered statistically significant. D) Immunoblotting analysis of AML cell lines (Molm-13, Molm-13/VR, U937) and PDX (2016-1, 2016-7) cells after 24h of treatment with CX-4945 and VEN alone or in combination. β-Actin was used as a loading control. Representative blots from two to three independent experiments were shown.

Article Snippet: CX-4945 (#HY-50855B) and venetoclax (#HY-15531) were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Western Blot, Quantitative RT-PCR, Control

Journal: bioRxiv

Article Title: CK2 inhibitor, CX-4945, enhances BH3 priming and promotes apoptosis of venetoclax-resistant AML by targeting antiapoptotic proteins

doi: 10.64898/2025.12.24.696284

Figure Lengend Snippet: A) Schematic depicting the treatment plan in AML patient-derived xenograft (PDX) mouse model. Similar scheme followed for cell line-derived xenograft (CDX) mice except the analysis after two-weeks of treatment. The NRG-S mice were irradiated and intravenously injected with AML cell lines Molm-13VR (0.25×10 6 cells/mouse), U937 (1×10 4 cells/mouse), and PDX 2016-7 (0.5×10 6 cells/mouse) and randomized into experimental groups (Vehicle, CX-4945, VEN, and CX+VEN Como). The drug treatment started after one week of cell transplantation and followed up for survival analysis or analysis after two-weeks of treatment (only for PDX). B-F) After two weeks of drug treatment, mice injected with PDX 2016-7 cells were sacrificed and spleen weight was recorded ( B ). The cells collected from spleens were analyzed by flow cytometry to analyze percent hCD45 positive cells ( C ). D-F) CBC analysis was performed on PDX 2016-7 mice after two-weeks of drug treatment by Hemavet analyzer. The data for B-F are presented as mean ± SD (n=7-8) and analyzed by one-way ANOVA (Tukey’s multiple comparisons test). *p<0.05, **p<0.01, ***p<0.001, and ****p<0.0001 indicates statistical significance, while ‘ns’ denotes ‘not significant’. G) Immunoblotting analysis of spleen cells collected from PDX 2016-7 mice after two-weeks of treatment with CX-4945, VEN, and CX+VEN combo. The data for three different experimental animals was presented. H-J) Kaplan-Meyer survival analysis of 2016-7 PDX ( H ), Molm-13/VR CDX ( I ), and U937 CDX ( J ) mice after treatment with CX-4945, VEN and CX+VEN combo. The overall median survival (MS) days for each experimental group of PDX and CDX mice were provided next to the legend. *p<0.05, **p<0.01 and ***p<0.001 by Gehan-Breslow-Wilcoxon test indicates statistical significance.

Article Snippet: CX-4945 (#HY-50855B) and venetoclax (#HY-15531) were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Derivative Assay, Irradiation, Injection, Transplantation Assay, Flow Cytometry, Western Blot

Journal: bioRxiv

Article Title: CK2 inhibitor, CX-4945, enhances BH3 priming and promotes apoptosis of venetoclax-resistant AML by targeting antiapoptotic proteins

doi: 10.64898/2025.12.24.696284

Figure Lengend Snippet: A) AML cells from patients were surfaced stained with different fluorescent labeled antibodies and analyzed for the expression of markers for LSCs (CD34, CD38, TIM3) and chemoresistance (CD47 and CD123) by flow cytometry. B) The basal level expression of CK2α, BCL-XL, and CK2 activity (p-CK2 substrate) was measured in primary AML cells by immunoblotting analysis. C) The basal level apoptosis and viability of primary AML patient cells was assessed after 12-24h of post-thaw using Annexin V & dead cell kit for Muse cell analyzer. The patient cells with high basal level apoptosis were omitted and the ones used for further testing were highlighted using red square box. D & E) AML patient cells were treated with CX-4945 and VEN alone and in combination for 24 h and surface stained for different cell surface markers along with annexin V. The relative apoptosis in bulk cells ( D ), LSCs (CD34+CD38−) and chemoresistant (CD47+CD123-) subpopulations were calculated by normalizing the percent apoptosis in vehicle treated cells ( E ). The data are presented as mean ± SEM (n=6-7) and analyzed via one-way ANOVA (Tukey’s multiple comparisons test). *p<0.05, **p<0.01, and ***p<0.001 considered as statistically significant.

Article Snippet: CX-4945 (#HY-50855B) and venetoclax (#HY-15531) were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Staining, Labeling, Expressing, Flow Cytometry, Activity Assay, Western Blot

Journal: bioRxiv

Article Title: CK2 inhibitor, CX-4945, enhances BH3 priming and promotes apoptosis of venetoclax-resistant AML by targeting antiapoptotic proteins

doi: 10.64898/2025.12.24.696284

Figure Lengend Snippet: Inhibition of CK2-mediated signaling through CX-4945 negatively affect cell survival pathways (PI3K/AKT/mTOR, NF-κB pathways) and anti-apoptotic proteins (MCL-1L and BCL-XL). Downregulation of anti-apoptotic BCL2 members (MCL-1L, BCL-XL) further enhance BCL-2 dependency and enhance VEN-mediated apoptosis in VR-AML cells. Overexpression of pro-apoptotic MCL-1ES isoform that can mediate mitochondrial depolarization independent of BAX/BAK contribute to potentiate VEN activity. Moreover, the transcriptome of VR-AML cells followed by CX-4945 and VEN combination treatment showed reversal of molecular gene signatures associated with VEN resistance and result in potentiated apoptosis in VR-AML cells. The signaling targeted by CX-4945 and VEN are represented by dotted lines. Created using BioRender.com .

Article Snippet: CX-4945 (#HY-50855B) and venetoclax (#HY-15531) were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Inhibition, Over Expression, Activity Assay

Journal: Cells

Article Title: Cancer Stem Cell and Aggressiveness Traits Are Promoted by Stable Endothelin-Converting Enzyme-1c in Glioblastoma Cells

doi: 10.3390/cells12030506

Figure Lengend Snippet: ECE1c K6R mutant was highly stable in GBM cells. Flag-tagged ECE1c WT - or ECE1c K6R -expressing U87MG ( A ), T98G ( B ), and U251 ( C ) cells were treated with 20 μg/mL cycloheximide (CHX) in the absence or presence of 25 μM silmitasertib for 6 h. ECE1c protein levels were evaluated using Western blots with an anti-Flag antibody, using β-actin as a loading control. Representative blots from three independent experiments are shown. Representative Western blots of ECE1c WT or ECE1c K6R and β-actin from three independent cell lines ( n = 3).

Article Snippet: Then, cells were incubated in the presence of 25 μM silmitasertib (a selective CK2 inhibitor, formerly known as CX-4945; Apexbio, Houston, TX, USA) or vehicle only (0.001% DMSO) for 6 h. Cells were then harvested and lysed, and the total protein was analyzed using Western blots.

Techniques: Mutagenesis, Expressing, Western Blot, Control

Journal: ASN NEURO

Article Title: Casein Kinase 2 Mediates HIV- and Opioid-Induced Pathologic Phosphorylation of TAR DNA Binding Protein 43 in the Basal Ganglia

doi: 10.1177/17590914231158218

Figure Lengend Snippet: Pathologic deposition of TDP-43 and casein kinase 2 (CK2) immunoreactivity in the basal ganglia of HIV+ individuals. (a,a’ and b,b’) Representative confocal images of TDP-43, casein kinase 1δ (CK1δ), pTDP-43, and CK2 immunofluorescence in HIV− (a,b) and HIV+ (a’,b’) basal ganglia. Note the overall paucity of CK1δ antigenicity and the increased nuclear-to-cytoplasmic redistribution of TDP-43 antigenicity in the neural cells of HIV+ individuals. (c) A high magnification (63×) image showing pTDP-43 and CK2 immunofluorescence colocalization in cells within post-mortem HIV+ basal ganglia.

Article Snippet: Cells were either treated with vehicle (DNase/RNase-free non-pyrogenic Ultrapure water), co-treated with Tat (100 nM) and morphine (500 nM), or a combination of Tat, morphine, and 0.5, 1, or 2 μM concentrations of the highly selective CK2 antagonist CX-4945 (#A11060, AdooQ Bioscience, Irvine, CA).

Techniques: Immunofluorescence

Journal: ASN NEURO

Article Title: Casein Kinase 2 Mediates HIV- and Opioid-Induced Pathologic Phosphorylation of TAR DNA Binding Protein 43 in the Basal Ganglia

doi: 10.1177/17590914231158218

Figure Lengend Snippet: Assessment of pathologic TDP-43 and casein kinase 2 (CK2) immunoreactivity in the basal ganglia of HIV− and HIV+ individuals. (a) Colocalization of pTDP-43, CK2, and DARPP32 immunoreactivity in cells of post-mortem HIV+ brains confirming the identity of the cells as medium spiny neurons. (b,c) Cytoplasmic TDP-43 (b) and pTDP-43 (c) fluorescence intensities were increased in HIV+ compared to HIV− basal ganglia. (d) Cytoplasmic-to-nuclear ratio of TDP-43 fluorescent intensity was increased in HIV+ tissues indicating mislocalization. (e) CK1δ fluorescent intensity was slightly above background levels and not changed in any of the groups evaluated. (f) Cytoplasmic CK2 intensity was increased in HIV+ compared to HIV− basal ganglia. Pearson correlations revealed no significant relationship between the nuclear (g) and cytoplasmic (h) intensities of pTDP-43 (pooled across groups) vs CK1δ fluorescence. On the contrary, nuclear (i) and cytoplasmic (j) intensities of pTDP-43 fluorescence (pooled across groups) correlated positively with CK2 fluorescence intensities. Mean intensity values indicate the mean fluorescence pixel intensities for each respective protein acquired in optical sections using confocal microscopy and analyzed using CellProfiler TM 6.1 software (see Materials and Methods 2.1.2). Data are expressed as the mean + the SEM ( n = 4-7/group). * indicates a significant difference by Student's t -test ( p < 0.05).

Article Snippet: Cells were either treated with vehicle (DNase/RNase-free non-pyrogenic Ultrapure water), co-treated with Tat (100 nM) and morphine (500 nM), or a combination of Tat, morphine, and 0.5, 1, or 2 μM concentrations of the highly selective CK2 antagonist CX-4945 (#A11060, AdooQ Bioscience, Irvine, CA).

Techniques: Fluorescence, Confocal Microscopy, Software

Journal: ASN NEURO

Article Title: Casein Kinase 2 Mediates HIV- and Opioid-Induced Pathologic Phosphorylation of TAR DNA Binding Protein 43 in the Basal Ganglia

doi: 10.1177/17590914231158218

Figure Lengend Snippet: Effects of morphine and Tat on the concentrations of TDP-43 and casein kinases in the mouse striatum. (a) Nuclear TDP-43 levels were not changed in any of the groups evaluated. (a,b) Alternatively, morphine exposure increased cytoplasmic TDP-43 levels (a), and nuclear and cytoplasmic levels of pTDP-43 (b), while Tat increased cytoplasmic pTDP-43 concentrations (b). (b) Post hoc comparisons showed differences in nuclear pTDP-43 between saline-treated (Tat+ and Tat−) mice vs mice co-exposed to Tat and morphine, and in cytoplasmic pTDP-43 levels between saline-treated Tat− mice vs mice co-exposed to Tat and morphine. (c) Representative immunoblots of cytoplasmic pTDP-43, TDP-43, CK1δ, CK2α, and CK2α’, and GAPDH. (d) The cytoplasmic-to-nuclear ratios of TDP-43 levels were not changed in any of the treatment groups. (e) Nuclear and cytoplasmic CK1δ levels were not changed in any of the groups evaluated. (f) When CK2 levels were evaluated, Tat increased nuclear CK2α levels, and either Tat or morphine alone increased cytoplasmic CK2α levels. (f) Post hoc comparisons of cytoplasmic CK2α levels showed differences between saline-treated Tat− mice vs Tat+ mice that were exposed to morphine. (g) Nuclear and cytoplasmic levels of the CK2α’ subunit were unaffected by Tat or morphine exposure. Data are expressed as the mean + the SEM ( n = 5-6/group). # indicates significant main effects by two-way ANOVA; * indicates planned contrasts with post hoc Tukey's test ( p < 0.05).

Article Snippet: Cells were either treated with vehicle (DNase/RNase-free non-pyrogenic Ultrapure water), co-treated with Tat (100 nM) and morphine (500 nM), or a combination of Tat, morphine, and 0.5, 1, or 2 μM concentrations of the highly selective CK2 antagonist CX-4945 (#A11060, AdooQ Bioscience, Irvine, CA).

Techniques: Saline, Western Blot

Journal: ASN NEURO

Article Title: Casein Kinase 2 Mediates HIV- and Opioid-Induced Pathologic Phosphorylation of TAR DNA Binding Protein 43 in the Basal Ganglia

doi: 10.1177/17590914231158218

Figure Lengend Snippet: Effects of morphine and Tat on the relationship between the concentrations of pTDP-43 and TDP-43 kinases. (a,b) No significant correlations were observed between levels of nuclear (a) or cytoplasmic (b) CK1δ and pTDP-43 . (c) No significant correlations were observed between levels of nuclear CK2α vs nuclear pTDP-43 . (d) By contrast, levels of cytoplasmic CK2α positively correlated with cytoplasmic pTDP-43 concentrations ( p < 0.05). (e,f) No significant correlations were observed between levels of nuclear (e) or cytoplasmic (f) CK2α’ ( Figure 3 g ) and pTDP-43 .

Article Snippet: Cells were either treated with vehicle (DNase/RNase-free non-pyrogenic Ultrapure water), co-treated with Tat (100 nM) and morphine (500 nM), or a combination of Tat, morphine, and 0.5, 1, or 2 μM concentrations of the highly selective CK2 antagonist CX-4945 (#A11060, AdooQ Bioscience, Irvine, CA).

Techniques:

Journal: ASN NEURO

Article Title: Casein Kinase 2 Mediates HIV- and Opioid-Induced Pathologic Phosphorylation of TAR DNA Binding Protein 43 in the Basal Ganglia

doi: 10.1177/17590914231158218

Figure Lengend Snippet: The effects of Tat and morphine on pTDP-43 levels and CK2 activity in striatal neurons in vitro . (a) Tat or morphine exposure did not affect the concentration of cytoplasmic TDP-43. (b) By contrast, Tat and morphine co-exposure significantly increased levels of cytoplasmic pTDP-43, and these increases in pTDP-43 were fully reversible by the CK2 antagonist (CX-4945) in a concentration-dependent manner. (c) Tat and morphine co-exposure increased CK2 enzymatic activity in cytoplasmic extracts of striatal neurons. (d) Representative immunoblots of cytoplasmic pTDP-43, TDP-43, and GAPDH. Symbols indicate planned contrasts with post hoc Tukey's test ( p < 0.05). $ vs the control group (no active treatment), $$ vs the control group at the 20 min reaction time, $$$ vs the control group at 40 min, ǂ vs Tat+/morphine group, ^ vs Tat+/morphine and Cx-4945 at 20 min, ^^ vs Tat+/morphine and Cx-4945 at 40 min, * vs the 0 min reaction time. Data are expressed as the mean + or ± the SEM ( n = 4 experiments/group).

Article Snippet: Cells were either treated with vehicle (DNase/RNase-free non-pyrogenic Ultrapure water), co-treated with Tat (100 nM) and morphine (500 nM), or a combination of Tat, morphine, and 0.5, 1, or 2 μM concentrations of the highly selective CK2 antagonist CX-4945 (#A11060, AdooQ Bioscience, Irvine, CA).

Techniques: Activity Assay, In Vitro, Concentration Assay, Western Blot, Control