Journal: Cell Death & Disease

Article Title: Prediction of venetoclax activity in precursor B-ALL by functional assessment of apoptosis signaling

doi: 10.1038/s41419-019-1801-0

Figure Lengend Snippet: Assaying the dependence of mitochondrial apoptosis signaling on different regulating molecules interrogating binding of specific BH3 peptides (BH3 profiling). a Experimental procedure, ALL cells are permeabilized, incubated with the respective peptide followed by detection of cytochrome c release. b Binding table showing interaction of the BH3-peptides (columns) with the respective apoptosis-regulating molecule (rows). Mitochondrial priming by c VEN, e BAD, and g BAD-HRK is significantly associated with ex vivo venetoclax sensitivity (linear regression; R 2 , correlation coefficient; p , significance) and d , f , h predictive for ex vivo response of ALL cells to venetoclax (ROC/receiver operating characteristic curve; AUC, area under the curve; p , significance)

Article Snippet: VEN was purchased from Chemietek (USA).

Techniques: Binding Assay, Incubation, Ex Vivo

Journal: Cell Death & Disease

Article Title: Prediction of venetoclax activity in precursor B-ALL by functional assessment of apoptosis signaling

doi: 10.1038/s41419-019-1801-0

Figure Lengend Snippet: Individual patient-derived BCP-ALL xenograft samples ( N = 12) were transplanted onto pairs of recipient mice and treated with either VEN or vehicle for 10 days. After treatment, mice were tightly monitored for onset of leukemia-related morbidity. a Survival times of leukemia bearing mice treated with VEN (gray bars) or vehicle (black bars) (left diagram) and corresponding VEN-induced survival differences (‘delta survival’, right diagram). b Association of direct VEN priming with preclinical VEN sensitivities of BCP-ALL in vivo (linear regression; R 2 , correlation coefficient; p , significance) and c predictive value of direct VEN priming for post-VEN survival of treated mice (ROC/receiver operating characteristic curve; AUC, area under the curve; p , significance). d Strong association of functional BCL-2 dependence (mitochondrial BAD-HRK priming) with in vivo VEN responses, and e high predictive value of BAD-HRK priming for survival of VEN treated mice. Preclinical in vivo VEN responses analyzed in larger treatment groups confirming f low (PDX13; N = 8 mice per group), g intermediate (PDX10; N = 10 mice per group), and h strong (PDX2, five mice per group) VEN sensitivity of the respective patient-derived xenograft leukemia observed in the preclinical trial (Kaplan–Meier analysis; p , significance by log-rank test; VEN, venetoclax; CTRL control)

Article Snippet: VEN was purchased from Chemietek (USA).

Techniques: Derivative Assay, In Vivo, Functional Assay, Control

Journal: Journal of Veterinary Internal Medicine

Article Title: Sensitivity of canine hematological cancers to BH3 mimetics

doi: 10.1111/jvim.16587

Figure Lengend Snippet: Summary cell viability data for normal canine CD3+ T and CD21+ B lymphocytes after 24 hours treatment with VEN and NAV, with viability normalized to DMSO control wells. Points and error bars represent the mean EC 50 ± SD. Viability of peripheral blood (A and B) from 8 dogs, and lymph node (C and D) aspirates from 5 dogs is shown

Article Snippet: Either VEN or NAV (AbbVie, North Chicago, Illinois), dissolved in DMSO to a final concentration of 1 mM, were added to cells at graded drug concentrations (2 nM to 10 μM).

Techniques: Control

Journal: Journal of Veterinary Internal Medicine

Article Title: Sensitivity of canine hematological cancers to BH3 mimetics

doi: 10.1111/jvim.16587

Figure Lengend Snippet: Estimated mean EC 50 ± SD for normal lymphocytes isolated from peripheral blood (8 dogs), and lymph node (5 dogs), after 24 hours treatment with VEN and NAV, calculated from dose–response curves presented in Figure 1

Article Snippet: Either VEN or NAV (AbbVie, North Chicago, Illinois), dissolved in DMSO to a final concentration of 1 mM, were added to cells at graded drug concentrations (2 nM to 10 μM).

Techniques: Isolation

Journal: Journal of Veterinary Internal Medicine

Article Title: Sensitivity of canine hematological cancers to BH3 mimetics

doi: 10.1111/jvim.16587

Figure Lengend Snippet: In vitro sensitivity of peripheral blood (PB) and lymph node (LN) derived B lymphocytes (A) and T lymphocytes (B) from individual dogs to VEN and for the same cells to NAV (C and D). Data show the log‐transformed EC 50 , with the horizontal line representing the mean and error bars showing SD. P ‐values derived from unpaired Student's t ‐test

Article Snippet: Either VEN or NAV (AbbVie, North Chicago, Illinois), dissolved in DMSO to a final concentration of 1 mM, were added to cells at graded drug concentrations (2 nM to 10 μM).

Techniques: In Vitro, Derivative Assay, Transformation Assay

Journal: Journal of Veterinary Internal Medicine

Article Title: Sensitivity of canine hematological cancers to BH3 mimetics

doi: 10.1111/jvim.16587

Figure Lengend Snippet: Paired comparisons of normal lymphocyte sensitivity to VEN and NAV. Connecting lines show sensitivity of lymphocytes from an individual dog treated under the same conditions, comparing different BH3 mimetics. Data represent log‐transformed EC 50 from individual dogs, with the horizontal line representing the mean and error bars showing SD. P ‐values derived from paired Student's t ‐test

Article Snippet: Either VEN or NAV (AbbVie, North Chicago, Illinois), dissolved in DMSO to a final concentration of 1 mM, were added to cells at graded drug concentrations (2 nM to 10 μM).

Techniques: Transformation Assay, Derivative Assay

Journal: Journal of Veterinary Internal Medicine

Article Title: Sensitivity of canine hematological cancers to BH3 mimetics

doi: 10.1111/jvim.16587

Figure Lengend Snippet: Representative dose response curve from 1 dog (dog 28) showing CD21+ B lymphocytes after 24 hours incubation with VEN or NAV. Data shown represent mean ± SD from triplicate wells and are normalized to the DMSO control

Article Snippet: Either VEN or NAV (AbbVie, North Chicago, Illinois), dissolved in DMSO to a final concentration of 1 mM, were added to cells at graded drug concentrations (2 nM to 10 μM).

Techniques: Incubation, Control

Journal: Journal of Veterinary Internal Medicine

Article Title: Sensitivity of canine hematological cancers to BH3 mimetics

doi: 10.1111/jvim.16587

Figure Lengend Snippet: Dot plot comparing the in vitro sensitivity of canine B and T cell cancers and unclassified leukemia to BH3 mimetics. Data represent the log transformed EC 50 with the mean (horizontal line) and SD (error bars) after 24 hours incubation with VEN or NAV. Groups were compared using 1‐way ANOVA. ** Indicates a P ‐value <.01

Article Snippet: Either VEN or NAV (AbbVie, North Chicago, Illinois), dissolved in DMSO to a final concentration of 1 mM, were added to cells at graded drug concentrations (2 nM to 10 μM).

Techniques: In Vitro, Transformation Assay, Incubation

Journal: Journal of Veterinary Internal Medicine

Article Title: Sensitivity of canine hematological cancers to BH3 mimetics

doi: 10.1111/jvim.16587

Figure Lengend Snippet: Dot plot comparing the in vitro sensitivity of canine B and T cell cancers, unclassified leukemia and normal lymphocytes to BH3 mimetics. Data represent the log transformed EC 50 with the mean (horizontal line) and SD (error bars) after 24 hours incubation with VEN or NAV. Groups were compared using 1‐way ANOVA. * Indicates a P ‐value <.05; ** indicates a P ‐value <.01; **** indicates a P ‐value <.0001

Article Snippet: Either VEN or NAV (AbbVie, North Chicago, Illinois), dissolved in DMSO to a final concentration of 1 mM, were added to cells at graded drug concentrations (2 nM to 10 μM).

Techniques: In Vitro, Transformation Assay, Incubation

Journal: Journal of Veterinary Internal Medicine

Article Title: Sensitivity of canine hematological cancers to BH3 mimetics

doi: 10.1111/jvim.16587

Figure Lengend Snippet: Dose response curves to VEN and to NAV showing mean viability of cells normalized to DMSO control ± SD of triplicate wells. (A) Dog (37) with advanced nodal marginal zone lymphoma. The EC 50 of CD21+ neoplastic nodal cells exceed 5 μM. (B) Dog (26) with multicentric large T cell lymphoma. The EC 50 of neoplastic CD3+ nodal T cells to VEN and NAV are 7.9 and 20.8 nM, respectively

Article Snippet: Either VEN or NAV (AbbVie, North Chicago, Illinois), dissolved in DMSO to a final concentration of 1 mM, were added to cells at graded drug concentrations (2 nM to 10 μM).

Techniques: Control

Journal: Blood Advances

Article Title: Venetoclax enhances the efficacy of therapeutic antibodies in B-cell malignancies by augmenting tumor cell phagocytosis

doi: 10.1182/bloodadvances.2022007364

Figure Lengend Snippet: Effects of venetoclax in DHL cells. (A-C) Trypan blue exclusion test of cell viability in CARNAVAL (A), WILL-2 (B), and Oci-Ly7 (C) cells treated with increasing concentrations of venetoclax (VEN) for up to 48 hours. Technical triplicates were counted per experiment. Graphs show results of 3 independent experiments (n = 3, SD). (D-F) Annexin V staining measured by flow cytometry in CARNAVAL (D), WILL-2 (E), and Oci-Ly7 (F) cells subjected to escalating concentrations of 1 nM to 1 µM VEN and DMSO for 12 hours. (G-I) Protein expression of Bcl-2 and apoptotic markers Caspase 3, 7, 9, cPARP, and total levels analyzed by Western blot in CARNAVAL (G), WILL-2 (H), and Oci-Ly7 (I) cells after treatment with VEN for 12 hours with concentrations as indicated. Tubulin served as a loading control. Quantification of Western blot was done with ImageJ. Intensities were calculated relative to tubulin and normalized to DMSO solvent control. N/A, not analyzable.

Article Snippet: VEN was purchased from LC Laboratories, navitoclax (NTX) from Selleckchem, N -benzyloxycarbonyl-Val-Ala-Asp( O -Me) fluoromethyl ketone (Z-VAD-FMK) from MedChemExpress, and AZD5991 from selleckchem.

Techniques: Staining, Flow Cytometry, Expressing, Western Blot, Control, Solvent

Journal: Blood Advances

Article Title: Blockage of BCL-XL overcomes venetoclax resistance across BCL2 + lymphoid malignancies irrespective of BIM status

doi: 10.1182/bloodadvances.2024012906

Figure Lengend Snippet: In vitro cytotoxic synergy induced by the combination of VEN and A1155463

Article Snippet: VEN-sensitive cell lines (HBL-2, MAVER-1, and UPF1H) and PDX cells (VFN-M1 and VFN-ALL6) were subcutaneously xenografted into immunodeficient NSG mice and treated with VEN (100 mg/kg per daily) until the development of VEN-R tumors.

Techniques: In Vitro

Journal: Blood Advances

Article Title: Blockage of BCL-XL overcomes venetoclax resistance across BCL2 + lymphoid malignancies irrespective of BIM status

doi: 10.1182/bloodadvances.2024012906

Figure Lengend Snippet: BAX belongs to key mediators of cytotoxicity triggered by the combination of VEN and A1155463 in BIM-deficient lymphoma and leukemia cells. (A) Western blot analysis of selected BCL2 family proteins in the tested lymphoma and leukemia cell lines. (B) Western blot confirmation of absence of BIM and/or BAK proteins in the HBL-2 and MAVER-1 lymphoma cell clones with K/O of BIM and BAK genes. (C) K/O of BIM in lymphoma cell lines (HBL-2 and MAVER-1 BIM K/O clones) resulted in increased VEN-R, whereas sensitivity to the combination was largely retained; sensitivity of wild-type (WT) cells is shown for comparison. (D) Immunoprecipitations (IPs) of 2 BIM-deficient cell lines (MINO and Z-138) exposed for 3 hours to A1155463 (1000 nM) or medium only with anti–BCL-XL and anti-BCL2 antibodies and subsequent western blot detection of BAX and BAK proteins bound on BCL-XL and BCL2. (E) Western blot confirmation of absence of BAX protein expression in HBL-2 cell clone with K/O of BAX gene. (F) Apoptosis essays after exposure to the defined VEN and A1155463 concentrations reveal that HBL-2 cells with BAX K/O are significantly more resistant to VEN and to the VEN + A1155463 combination than HBL-2 cells with BAK K/O.

Article Snippet: VEN-sensitive cell lines (HBL-2, MAVER-1, and UPF1H) and PDX cells (VFN-M1 and VFN-ALL6) were subcutaneously xenografted into immunodeficient NSG mice and treated with VEN (100 mg/kg per daily) until the development of VEN-R tumors.

Techniques: Western Blot, Clone Assay, Comparison, Expressing

Journal: Blood Advances

Article Title: Blockage of BCL-XL overcomes venetoclax resistance across BCL2 + lymphoid malignancies irrespective of BIM status

doi: 10.1182/bloodadvances.2024012906

Figure Lengend Snippet: BCL-XL belongs to key mediators of sensitivity/resistance to VEN-induced apoptosis. (A) Representative western blot analysis confirms absence of BCL-XL protein expression in the HBL-2 and MAVER-1 lymphoma clones with K/O of BCL-XL gene. (B) Calculated lethal dose 50 (LD 50 ) for A1155463 and VEN in the HBL-2 and MAVER-1 cell clones with BCL-XL K/O compared with the WT lymphoma cell lines. (C) Western blot analysis confirms doxycycline-induced dose-dependent re-expression of BCL-XL protein in HBL-2 BCL-XL K/O clone with transgenic re-expression of BCL-XL (designated BCL-XL K/O R); expression of BCL-XL in the WT HBL-2 cell line is shown for comparison. (D) Extent of the doxycycline-induced transgenic re-expression of BCL-XL in the HBL-2 BCL-XL K/O R clone under doxycycline promotor negatively correlates with sensitivity to VEN; sensitivity to HBL-2 WT cell line and HBL-2 BCL-XL K/O clone is shown for comparison. DOX, doxycycline.

Article Snippet: VEN-sensitive cell lines (HBL-2, MAVER-1, and UPF1H) and PDX cells (VFN-M1 and VFN-ALL6) were subcutaneously xenografted into immunodeficient NSG mice and treated with VEN (100 mg/kg per daily) until the development of VEN-R tumors.

Techniques: Western Blot, Expressing, Clone Assay, Transgenic Assay, Comparison

Journal: Blood Advances

Article Title: Blockage of BCL-XL overcomes venetoclax resistance across BCL2 + lymphoid malignancies irrespective of BIM status

doi: 10.1182/bloodadvances.2024012906

Figure Lengend Snippet: Upregulation of BCL-XL induced by microenvironmental factors belongs to key mediators of VEN resistance in vitro. (A) Western blot analysis of BCL-XL protein after 24-hour coculture of the tested lymphoma cell lines (HBL-2, MAVER-1, UPF1H, and MINO) with HS5 CD40L feeder cells with stable transgenic expression of human CD40 ligand (HS5 CD40L). (B) Bar charts showing the differences of normalized expression levels of BCL2L1/BCL-XL in the tested WT lymphoma cell lines (blue bars) and after 24-hour coculture with HS5 CD40L feeder cells (red bars). (C) Coculture of HBL-2 and MAVER-1 WT lymphoma cell lines with HS5 CD40L feeder cells for 24 hours resulted in VEN resistance as measured by numbers of apoptotic cells after exposure to VEN (10 and 100 ng/mL); in contrast, coculture of HBL-2 and MAVER-1 BCL-XL K/O clones with HS5 CD40L did not change their sensitivity to VEN. (D) Combined treatment of the WT lymphoma cell lines cocultured for 24 hours on HS5 CD40L feeder cells with the combination of VEN (10 and 100 nM) and A1155463 (1000 nM) overcame the microenvironment-induced VEN resistance.

Article Snippet: VEN-sensitive cell lines (HBL-2, MAVER-1, and UPF1H) and PDX cells (VFN-M1 and VFN-ALL6) were subcutaneously xenografted into immunodeficient NSG mice and treated with VEN (100 mg/kg per daily) until the development of VEN-R tumors.

Techniques: In Vitro, Western Blot, Transgenic Assay, Expressing, Clone Assay

Journal: Blood Advances

Article Title: Blockage of BCL-XL overcomes venetoclax resistance across BCL2 + lymphoid malignancies irrespective of BIM status

doi: 10.1182/bloodadvances.2024012906

Figure Lengend Snippet: Sequestration of BIM to upregulated BCL-XL belongs to hallmarks of acquired VEN resistance. (A-E) Western blot analysis of selected BCL2 family proteins (MCL1, BCL-XL, BCL2, BAK, and BIM) in cell lysates isolated from the subcutaneous cell line–based xenograft (CDX) tumors (HBL-2, MAVER-1, and UPF1H), and subcutaneous patient-derived xenograft (PDX) tumors (VFN-M1 and VFN-ALL6) obtained from the untreated animals (CTRL) and from the animals with acquired VEN resistance. (F-H) IP of the selected CDX or PDX tumors with anti-BIM antibody and subsequent detection of BCL-XL and BCL2 bound on BIM; 1 CDX model (UPF1H) and 2 PDX models (VFN-M1 and VFN-ALL6) were analyzed; the mice were euthanized and tumors analyzed 24 hours after the last dose of VEN.

Article Snippet: VEN-sensitive cell lines (HBL-2, MAVER-1, and UPF1H) and PDX cells (VFN-M1 and VFN-ALL6) were subcutaneously xenografted into immunodeficient NSG mice and treated with VEN (100 mg/kg per daily) until the development of VEN-R tumors.

Techniques: Western Blot, Isolation, Derivative Assay

Journal: Blood Advances

Article Title: Blockage of BCL-XL overcomes venetoclax resistance across BCL2 + lymphoid malignancies irrespective of BIM status

doi: 10.1182/bloodadvances.2024012906

Figure Lengend Snippet: Cotargeting of BCL2 and BCL-XL with VEN and A1155463 is synthetically lethal on a panel of PDX models derived from patients with MCL, ALL, and DLBCL. (A) Cotargeting of BCL2 and BCL-XL with VEN and A1155463 was tested on a panel of 9 PDX models including MCL (VFN-M1, VFN-M5R1, and VFN-M16), ALL (VFN-ALL1, VFN-ALL6, and VFN-ALL7), and DLBCL (VFN-D1, VFN-D9, and VFN-D20); x-axis shows days from initiation of therapy; y-axis shows calculated tumor volumes ± standard deviations of PDX tumors; therapy was administered on days 1 to 4 and 8 to 12 (for details, see “Methods”). (B) Cotargeting of BCL2 and BCL-XL with VEN and A1155463 was effective even in the PDX models with acquired VEN resistance (VFN-M1 VEN-R and VFN-ALL6 VEN-R); VEN-R PDX tumors were derived from animals, in which originally VEN-sensitive PDX tumors grew on continued VEN therapy; such VEN-R PDX tumors were re-engrafted into secondary recipients and subjected to retreatment; each cohort of mice comprised 6 animals.

Article Snippet: VEN-sensitive cell lines (HBL-2, MAVER-1, and UPF1H) and PDX cells (VFN-M1 and VFN-ALL6) were subcutaneously xenografted into immunodeficient NSG mice and treated with VEN (100 mg/kg per daily) until the development of VEN-R tumors.

Techniques: Derivative Assay

Journal: Cell Death Discovery

Article Title: Venlafaxine antagonizes the noradrenaline-promoted colon cancer progression by inhibiting the norepinephrine transporter

doi: 10.1038/s41420-023-01447-5

Figure Lengend Snippet: Colon cancer cells were treated with control, venlafaxine (VEN) (10 μM), NE (10 μM), and drug combination [NE (10 μM) + VEN (10 μM)]. A Western blotting and B Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) assay were used to detect the changes in NET protein and mRNA. The band intensities of NET relative to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were quantified and normalized to the Ctrl. group. The quantification graphs of normalized band intensities of different replicates were shown in Fig. . C Colon cancer cells were transfected with siNET, sihNET, or siNC separately to knock down the expression of NET. After 24 h, they were treated with 10 μM of NE and incubated further for 24 h and 48 h. The effects of NE/siNET on colon cancer cell proliferation were determined by 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) assay. D Western blotting and E qRT-PCR assay were performed and revealed the knockdown of NET inhibited the NE-increased VEGF protein,VEGF mRNA, and Akt activation in colon cancer cells. The band intensities of VEGF, pAkt, and Akt relative to GAPDH were quantified and normalized to the siNC group. The quantification graphs of normalized band intensities of different replicates were shown in Fig. . Data are expressed as mean ± SD, * P < 0.05, ** P < 0.01, or *** P < 0.001 in ( B ), ( C ) and ( E ).

Article Snippet: Norepinephrine (NE) was purchased from Sigma (Sigma, USA), and VEN from Cayman Chemical (Cayman Chemical, USA).

Techniques: Control, Western Blot, Reverse Transcription, Polymerase Chain Reaction, Quantitative RT-PCR, Transfection, Knockdown, Expressing, Incubation, MTT Assay, Activation Assay

Journal: Cell Death Discovery

Article Title: Venlafaxine antagonizes the noradrenaline-promoted colon cancer progression by inhibiting the norepinephrine transporter

doi: 10.1038/s41420-023-01447-5

Figure Lengend Snippet: Colon cancer cells were treated with control, venlafaxine (VEN) (10 μM), NE (10 μM), and drug combination [NE (10 μM) + VEN (10 μM)] separately. A Western blotting and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) assay were performed and revealed that NE reduced the protein level of PPP2R1A, while VEN increased it; there was no influence on PPP2R1A mRNA levels with NE/VEN treatment. The band intensities of PPP2R1A relative to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were quantified and normalized to the Ctrl. group. The quantification graphs of normalized band intensities of different replicates were shown in Fig. . B Colon cancer cells were transfected with siNET, sihNET, or siNC separately. After 24 h, they were treated with 10 μM of NE and incubated further. Western blotting and qRT-PCR assay were performed and revealed that the knockdown of NET caused an increase in NE-reduced PPP2R1A protein expression; there was no influence on PPP2R1A mRNA levels with NE/ siNET treatment. The band intensities of PPP2R1A relative to GAPDH were quantified and normalized to the siNC group. The quantification graphs of normalized band intensities of different replicates were shown in Fig. . C The plasmids overexpressing NET and PPP2R1A were constructed, and short interfering RNAs against PPP2R1A were synthesized. They were transfected into CT26 cells separately. Western blotting and qRT-PCR were used to detect the changes in NET and PPP2R1A. The band intensities of NET and PPP2R1A relative to GAPDH were quantified and normalized to the ovCtrl./siNC group. The quantification graphs of normalized band intensities of different replicates were shown in Fig. . D Coimmunoprecipitation (Co-IP) assay was performed and revealed the existence of NET-PPP2R1A interaction. Data are expressed as mean ± SD, ** P < 0.01, or *** P < 0.001 in C .

Article Snippet: Norepinephrine (NE) was purchased from Sigma (Sigma, USA), and VEN from Cayman Chemical (Cayman Chemical, USA).

Techniques: Control, Western Blot, Reverse Transcription, Polymerase Chain Reaction, Quantitative RT-PCR, Transfection, Incubation, Knockdown, Expressing, Construct, Synthesized, Co-Immunoprecipitation Assay

Journal: Cell Death Discovery

Article Title: Venlafaxine antagonizes the noradrenaline-promoted colon cancer progression by inhibiting the norepinephrine transporter

doi: 10.1038/s41420-023-01447-5

Figure Lengend Snippet: A CT26 cells were transfected with plasmids of ovCtrl., ovPPP2R1A, or ovNET separately. Twenty-four hours later, they were treated with NE or not and incubated further for 48 h. Western blotting was used to detect the changes in PPP2R1A, pAkt, Akt, and VEGF proteins. The quantification graphs of normalized band intensities of different replicates were shown in Fig. . B CT26 cells were transfected with short interfering RNAs of siNC, siPPP2R1A-1, or siPPP2R1A-2 separately. Twenty-four hours later, they were treated with venlafaxine (VEN), NE, and NE + VEN, or not and incubated further for 48 h. Western blotting was used to detect the changes in PPP2R1A, pAkt, Akt, and VEGF proteins. The band intensities of PPP2R1A, pAkt, Akt, and VEGF relative to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were quantified and normalized to the ovCtrl./siNC group. The quantification graphs of normalized band intensities of different replicates were shown in Fig. . Experiments were repeated three independent times with reproducible results.

Article Snippet: Norepinephrine (NE) was purchased from Sigma (Sigma, USA), and VEN from Cayman Chemical (Cayman Chemical, USA).

Techniques: Transfection, Incubation, Western Blot

Journal: Cell Death Discovery

Article Title: Venlafaxine antagonizes the noradrenaline-promoted colon cancer progression by inhibiting the norepinephrine transporter

doi: 10.1038/s41420-023-01447-5

Figure Lengend Snippet: By binding with beta-adrenergic receptor (b-AR), norepinephrine (NE) induces Akt activation, VEGF expression, cell proliferation, angiogenesis, and cancer progression in human colorectal cancer (CRC) cells. NE also increases the expression of NE transporter (NET). Venlafaxine (VEN) can antagonize the above effects of NE by inhibiting the NE-increased NET expression, which is related to the interaction of NET and protein phosphatase 2 scaffold subunit alpha (PPP2R1A).

Article Snippet: Norepinephrine (NE) was purchased from Sigma (Sigma, USA), and VEN from Cayman Chemical (Cayman Chemical, USA).

Techniques: Binding Assay, Activation Assay, Expressing