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human neuroblastoma cell line sh sy5y  (ATCC)


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    ATCC human neuroblastoma cell line sh sy5y
    Effect of BDNF treatment on the neurite outgrowth parameters <t>in</t> <t>SH-SY5Y</t> cells Differentiated SH-SY5Y cells were treated with 10 ng/mL, 50 ng/mL, or 100 ng/mL of BDNF for 48h. (A) Cells were stained for ßIII-tubulin (red) and DAPI (blue). All images were captured 48h post-treatment at 20× magnification on the Agilent BioTek Cytation 5 instrument. Image analysis was performed with the Gen5 software Neurite Outgrowth module where the soma mask overlay is displayed in blue and the neurite mask overlay in yellow. (B) Average neurite length, (C) average neurite branches, (D) average neurite count depending on BDNF concentration, and (E) neurite thickness depending on BDNF concentration. Scale bar: 50 μm. Each data set represents N=2 independent experiments with 4-6 replicates per condition and with 5-9 images per well. The points represent the mean ± SEM for each condition. One-way ANOVA and post hoc Dunnett’s multiple comparison tests. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001. BDNF: Brain-derived neurotrophic factor.
    Human Neuroblastoma Cell Line Sh Sy5y, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 8995 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/human+neuroblastoma+cell+lines/pmc13068831-99-2-9?v=ATCC
    Average 99 stars, based on 8995 article reviews
    human neuroblastoma cell line sh sy5y - by Bioz Stars, 2026-07
    99/100 stars

    Images

    1) Product Images from "Protocol for automated quantification of neuronal cells using the Agilent BioTek Cytation system and Gen5 neurite outgrowth module"

    Article Title: Protocol for automated quantification of neuronal cells using the Agilent BioTek Cytation system and Gen5 neurite outgrowth module

    Journal: STAR Protocols

    doi: 10.1016/j.xpro.2026.104485

    Effect of BDNF treatment on the neurite outgrowth parameters in SH-SY5Y cells Differentiated SH-SY5Y cells were treated with 10 ng/mL, 50 ng/mL, or 100 ng/mL of BDNF for 48h. (A) Cells were stained for ßIII-tubulin (red) and DAPI (blue). All images were captured 48h post-treatment at 20× magnification on the Agilent BioTek Cytation 5 instrument. Image analysis was performed with the Gen5 software Neurite Outgrowth module where the soma mask overlay is displayed in blue and the neurite mask overlay in yellow. (B) Average neurite length, (C) average neurite branches, (D) average neurite count depending on BDNF concentration, and (E) neurite thickness depending on BDNF concentration. Scale bar: 50 μm. Each data set represents N=2 independent experiments with 4-6 replicates per condition and with 5-9 images per well. The points represent the mean ± SEM for each condition. One-way ANOVA and post hoc Dunnett’s multiple comparison tests. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001. BDNF: Brain-derived neurotrophic factor.
    Figure Legend Snippet: Effect of BDNF treatment on the neurite outgrowth parameters in SH-SY5Y cells Differentiated SH-SY5Y cells were treated with 10 ng/mL, 50 ng/mL, or 100 ng/mL of BDNF for 48h. (A) Cells were stained for ßIII-tubulin (red) and DAPI (blue). All images were captured 48h post-treatment at 20× magnification on the Agilent BioTek Cytation 5 instrument. Image analysis was performed with the Gen5 software Neurite Outgrowth module where the soma mask overlay is displayed in blue and the neurite mask overlay in yellow. (B) Average neurite length, (C) average neurite branches, (D) average neurite count depending on BDNF concentration, and (E) neurite thickness depending on BDNF concentration. Scale bar: 50 μm. Each data set represents N=2 independent experiments with 4-6 replicates per condition and with 5-9 images per well. The points represent the mean ± SEM for each condition. One-way ANOVA and post hoc Dunnett’s multiple comparison tests. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001. BDNF: Brain-derived neurotrophic factor.

    Techniques Used: Staining, Software, Concentration Assay, Comparison, Derivative Assay

    Effect of rotenone treatment on the neurite outgrowth parameters in SH-SY5Y cells Differentiated SH-SY5Y cells were treated with 0.05 μM, 0.1 μM or 0.5 μM of rotenone for 48h. (A) Cells were stained for ß3 tubulin (red color) and DAPI (blue color). All images were captured 48h post-treatment at 20× magnification on the Agilent BioTek Cytation 5 instrument. Image analysis was performed with the Gen5 software Neurite Outgrowth module where the soma mask overlay is displayed in blue and the neurite mask overlay in yellow. (B) Average neurite length, (C) average neurite branches, (D) average neurite count, and (E) neurite thickness depending on rotenone concentration. Scale bar: 50 μm. Each data set represents N=2 independent experiments with 4-6 replicates per condition and with 5-9 pictures per well. The points represent the mean ± SEM for each condition. One-way ANOVA and post hoc Dunnett’s multiple comparison tests. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.
    Figure Legend Snippet: Effect of rotenone treatment on the neurite outgrowth parameters in SH-SY5Y cells Differentiated SH-SY5Y cells were treated with 0.05 μM, 0.1 μM or 0.5 μM of rotenone for 48h. (A) Cells were stained for ß3 tubulin (red color) and DAPI (blue color). All images were captured 48h post-treatment at 20× magnification on the Agilent BioTek Cytation 5 instrument. Image analysis was performed with the Gen5 software Neurite Outgrowth module where the soma mask overlay is displayed in blue and the neurite mask overlay in yellow. (B) Average neurite length, (C) average neurite branches, (D) average neurite count, and (E) neurite thickness depending on rotenone concentration. Scale bar: 50 μm. Each data set represents N=2 independent experiments with 4-6 replicates per condition and with 5-9 pictures per well. The points represent the mean ± SEM for each condition. One-way ANOVA and post hoc Dunnett’s multiple comparison tests. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.

    Techniques Used: Staining, Software, Concentration Assay, Comparison

    Impact of the APP and P301L mutation on the neurite outgrowth capacity of SH-SY5Y cells The control condition corresponds to the native cells. The APP condition corresponds to the cells stably overexpressing the Amyloid Precursor Protein (APP). The P301L condition corresponds to the cells stably overexpressing the P301L-Tau mutation. The differentiated cells were treated with 50 ng/mL of BDNF for 48h. (A) Cells were stained for ß3 tubulin (red) and DAPI (blue). All images were captured 48h post-treatment at 20× magnification on the Agilent BioTek Cytation 5 instrument. Image analysis was performed with the Gen5 software Neurite Outgrowth module where the soma mask overlay is displayed in blue and the neurite mask overlay in yellow. (B) Average neurite length, (C) average neurite branches, (D) average neurite count, and (E) neurite thickness in Control, APP, and P301L cells. Scale bar: 50 μm. The boxes represent the median (full line) and the mean (“+” symbol); the whiskers represent the minimum and maximum values. Each data set represents N=2 independent experiments with 4-6 replicates per condition. One-way ANOVA and post hoc Dunnett’s multiple comparison tests. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.
    Figure Legend Snippet: Impact of the APP and P301L mutation on the neurite outgrowth capacity of SH-SY5Y cells The control condition corresponds to the native cells. The APP condition corresponds to the cells stably overexpressing the Amyloid Precursor Protein (APP). The P301L condition corresponds to the cells stably overexpressing the P301L-Tau mutation. The differentiated cells were treated with 50 ng/mL of BDNF for 48h. (A) Cells were stained for ß3 tubulin (red) and DAPI (blue). All images were captured 48h post-treatment at 20× magnification on the Agilent BioTek Cytation 5 instrument. Image analysis was performed with the Gen5 software Neurite Outgrowth module where the soma mask overlay is displayed in blue and the neurite mask overlay in yellow. (B) Average neurite length, (C) average neurite branches, (D) average neurite count, and (E) neurite thickness in Control, APP, and P301L cells. Scale bar: 50 μm. The boxes represent the median (full line) and the mean (“+” symbol); the whiskers represent the minimum and maximum values. Each data set represents N=2 independent experiments with 4-6 replicates per condition. One-way ANOVA and post hoc Dunnett’s multiple comparison tests. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.

    Techniques Used: Mutagenesis, Control, Stable Transfection, Staining, Software, Comparison



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    Image Search Results


    Effect of BDNF treatment on the neurite outgrowth parameters in SH-SY5Y cells Differentiated SH-SY5Y cells were treated with 10 ng/mL, 50 ng/mL, or 100 ng/mL of BDNF for 48h. (A) Cells were stained for ßIII-tubulin (red) and DAPI (blue). All images were captured 48h post-treatment at 20× magnification on the Agilent BioTek Cytation 5 instrument. Image analysis was performed with the Gen5 software Neurite Outgrowth module where the soma mask overlay is displayed in blue and the neurite mask overlay in yellow. (B) Average neurite length, (C) average neurite branches, (D) average neurite count depending on BDNF concentration, and (E) neurite thickness depending on BDNF concentration. Scale bar: 50 μm. Each data set represents N=2 independent experiments with 4-6 replicates per condition and with 5-9 images per well. The points represent the mean ± SEM for each condition. One-way ANOVA and post hoc Dunnett’s multiple comparison tests. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001. BDNF: Brain-derived neurotrophic factor.

    Journal: STAR Protocols

    Article Title: Protocol for automated quantification of neuronal cells using the Agilent BioTek Cytation system and Gen5 neurite outgrowth module

    doi: 10.1016/j.xpro.2026.104485

    Figure Lengend Snippet: Effect of BDNF treatment on the neurite outgrowth parameters in SH-SY5Y cells Differentiated SH-SY5Y cells were treated with 10 ng/mL, 50 ng/mL, or 100 ng/mL of BDNF for 48h. (A) Cells were stained for ßIII-tubulin (red) and DAPI (blue). All images were captured 48h post-treatment at 20× magnification on the Agilent BioTek Cytation 5 instrument. Image analysis was performed with the Gen5 software Neurite Outgrowth module where the soma mask overlay is displayed in blue and the neurite mask overlay in yellow. (B) Average neurite length, (C) average neurite branches, (D) average neurite count depending on BDNF concentration, and (E) neurite thickness depending on BDNF concentration. Scale bar: 50 μm. Each data set represents N=2 independent experiments with 4-6 replicates per condition and with 5-9 images per well. The points represent the mean ± SEM for each condition. One-way ANOVA and post hoc Dunnett’s multiple comparison tests. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001. BDNF: Brain-derived neurotrophic factor.

    Article Snippet: The native human neuroblastoma cell line SH-SY5Y comes from ATCC (CRL-2266).

    Techniques: Staining, Software, Concentration Assay, Comparison, Derivative Assay

    Effect of rotenone treatment on the neurite outgrowth parameters in SH-SY5Y cells Differentiated SH-SY5Y cells were treated with 0.05 μM, 0.1 μM or 0.5 μM of rotenone for 48h. (A) Cells were stained for ß3 tubulin (red color) and DAPI (blue color). All images were captured 48h post-treatment at 20× magnification on the Agilent BioTek Cytation 5 instrument. Image analysis was performed with the Gen5 software Neurite Outgrowth module where the soma mask overlay is displayed in blue and the neurite mask overlay in yellow. (B) Average neurite length, (C) average neurite branches, (D) average neurite count, and (E) neurite thickness depending on rotenone concentration. Scale bar: 50 μm. Each data set represents N=2 independent experiments with 4-6 replicates per condition and with 5-9 pictures per well. The points represent the mean ± SEM for each condition. One-way ANOVA and post hoc Dunnett’s multiple comparison tests. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.

    Journal: STAR Protocols

    Article Title: Protocol for automated quantification of neuronal cells using the Agilent BioTek Cytation system and Gen5 neurite outgrowth module

    doi: 10.1016/j.xpro.2026.104485

    Figure Lengend Snippet: Effect of rotenone treatment on the neurite outgrowth parameters in SH-SY5Y cells Differentiated SH-SY5Y cells were treated with 0.05 μM, 0.1 μM or 0.5 μM of rotenone for 48h. (A) Cells were stained for ß3 tubulin (red color) and DAPI (blue color). All images were captured 48h post-treatment at 20× magnification on the Agilent BioTek Cytation 5 instrument. Image analysis was performed with the Gen5 software Neurite Outgrowth module where the soma mask overlay is displayed in blue and the neurite mask overlay in yellow. (B) Average neurite length, (C) average neurite branches, (D) average neurite count, and (E) neurite thickness depending on rotenone concentration. Scale bar: 50 μm. Each data set represents N=2 independent experiments with 4-6 replicates per condition and with 5-9 pictures per well. The points represent the mean ± SEM for each condition. One-way ANOVA and post hoc Dunnett’s multiple comparison tests. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.

    Article Snippet: The native human neuroblastoma cell line SH-SY5Y comes from ATCC (CRL-2266).

    Techniques: Staining, Software, Concentration Assay, Comparison

    Impact of the APP and P301L mutation on the neurite outgrowth capacity of SH-SY5Y cells The control condition corresponds to the native cells. The APP condition corresponds to the cells stably overexpressing the Amyloid Precursor Protein (APP). The P301L condition corresponds to the cells stably overexpressing the P301L-Tau mutation. The differentiated cells were treated with 50 ng/mL of BDNF for 48h. (A) Cells were stained for ß3 tubulin (red) and DAPI (blue). All images were captured 48h post-treatment at 20× magnification on the Agilent BioTek Cytation 5 instrument. Image analysis was performed with the Gen5 software Neurite Outgrowth module where the soma mask overlay is displayed in blue and the neurite mask overlay in yellow. (B) Average neurite length, (C) average neurite branches, (D) average neurite count, and (E) neurite thickness in Control, APP, and P301L cells. Scale bar: 50 μm. The boxes represent the median (full line) and the mean (“+” symbol); the whiskers represent the minimum and maximum values. Each data set represents N=2 independent experiments with 4-6 replicates per condition. One-way ANOVA and post hoc Dunnett’s multiple comparison tests. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.

    Journal: STAR Protocols

    Article Title: Protocol for automated quantification of neuronal cells using the Agilent BioTek Cytation system and Gen5 neurite outgrowth module

    doi: 10.1016/j.xpro.2026.104485

    Figure Lengend Snippet: Impact of the APP and P301L mutation on the neurite outgrowth capacity of SH-SY5Y cells The control condition corresponds to the native cells. The APP condition corresponds to the cells stably overexpressing the Amyloid Precursor Protein (APP). The P301L condition corresponds to the cells stably overexpressing the P301L-Tau mutation. The differentiated cells were treated with 50 ng/mL of BDNF for 48h. (A) Cells were stained for ß3 tubulin (red) and DAPI (blue). All images were captured 48h post-treatment at 20× magnification on the Agilent BioTek Cytation 5 instrument. Image analysis was performed with the Gen5 software Neurite Outgrowth module where the soma mask overlay is displayed in blue and the neurite mask overlay in yellow. (B) Average neurite length, (C) average neurite branches, (D) average neurite count, and (E) neurite thickness in Control, APP, and P301L cells. Scale bar: 50 μm. The boxes represent the median (full line) and the mean (“+” symbol); the whiskers represent the minimum and maximum values. Each data set represents N=2 independent experiments with 4-6 replicates per condition. One-way ANOVA and post hoc Dunnett’s multiple comparison tests. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.

    Article Snippet: The native human neuroblastoma cell line SH-SY5Y comes from ATCC (CRL-2266).

    Techniques: Mutagenesis, Control, Stable Transfection, Staining, Software, Comparison

    Biomimetic NCM/IMP@HMSNs delivery system protective effects against OGD/R induced injury in SH-SY5Y cells. ( A ) Uptake Behavior of NCM/IMP@HMSNs in SH-SY5Y Cells. ( B ) Western Blot of the Effects of Each Treatment Group on the Nrf-2/ARE/Keap1 Pathway in SH-SY5Y Cells After OGD/R. ( C–E ) Bar Graph of the Relative Expression Levels of Keap1, Nrf2 and HO-1 Protein Among Different Groups. ( F ) Cell viability. ( G and H ) Nrf2 Protein Expressions in Each Group after ML385 Treatment. n.s., no significance, * p < 0.05, ** p < 0.01, vs. OGD/R group; && p < 0.01, vs. NCM/IMP@HMSNs group.

    Journal: International Journal of Nanomedicine

    Article Title: Neutrophil-Membrane Biomimetic Hollow Mesoporous Silica Nanoparticles for Targeted Delivery of Imperatorin to Alleviate Cerebral Ischemia–Reperfusion Injury via Nrf2/ARE/Keap1 Pathway

    doi: 10.2147/IJN.S592970

    Figure Lengend Snippet: Biomimetic NCM/IMP@HMSNs delivery system protective effects against OGD/R induced injury in SH-SY5Y cells. ( A ) Uptake Behavior of NCM/IMP@HMSNs in SH-SY5Y Cells. ( B ) Western Blot of the Effects of Each Treatment Group on the Nrf-2/ARE/Keap1 Pathway in SH-SY5Y Cells After OGD/R. ( C–E ) Bar Graph of the Relative Expression Levels of Keap1, Nrf2 and HO-1 Protein Among Different Groups. ( F ) Cell viability. ( G and H ) Nrf2 Protein Expressions in Each Group after ML385 Treatment. n.s., no significance, * p < 0.05, ** p < 0.01, vs. OGD/R group; && p < 0.01, vs. NCM/IMP@HMSNs group.

    Article Snippet: The human neuroblastoma cell line SH-SY5Y was purchased from Wuhan Procell Life Science & Technology Co., Ltd. SH-SY5Y cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin (P/S) mixture, and maintained in a cell incubator at 37°C with 5% CO 2 .

    Techniques: Western Blot, Expressing

    (A) General Synthesis of QW-5–70. i). 60% NaH, SEMCl, THF, 0–25 °C; ii). n-BuLi, 3,4,5-trimethoxybenzaldehyde, THF, −78 °C; iii). Dess-Martin, CH 2 Cl 2 , r.t.; iv)1-(phenylsulfonyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole, Pd(PPh 3 ) 4 , Na 2 CO 3 , 1,4-dioxane/H 2 O (v/v = 2/1), reflux; v). Pd(OAC) 2 , PPh 3 , K 2 CO 3 , n-butanol, reflux; vi). 37% HCl, MeOH, reflux. (B) QW-5–70 induced tubulin depolymerization. Colchicine and paclitaxel were included as reference compounds. (C) SPR sensorgrams of QW-5–70 and colchicine binding to tubulin. (mean ± SEM, n = 3) (D) EBI competition assays. The upper β-tubulin band represents native β-tubulin, while the lower band corresponds to the EBI–β-tubulin adduct. GAPDH served as a loading control. (E) Immunoblot analysis of soluble and polymerized β-tubulin in PC-3 cells following treatment with QW-5–70, colchicine, or paclitaxel (20 nM). Cells were fractionated into soluble and polymerized tubulin pools, which were analyzed by immunoblotting. Ponceau S staining and GAPDH were used as loading controls for polymerized and soluble fractions, respectively. Quantification of polymerized β-tubulin expressed as a percentage of total β-tubulin (soluble + polymerized). (mean ± SEM, n=3). (F) Representative immunofluorescence images of interphase and mitotic SK-N-BE(2)-C, NB-1691, and PC-3 cells treated with QW-5–70 (2 nM and 5 nM) for 24 h. Microtubules were stained with anti-α-tubulin antibodies (red), and nuclei were counterstained with DAPI (blue). Scale bar = 50 μm (n=3). (G) High-resolution X-ray crystal structure of the sT2R complex with QW-5–70. Left panel: detailed interactions of QW-5–70 (green sticks) within the colchicine-binding site are shown. Bound water molecules are represented as red spheres, and hydrogen bonds are indicated by dashed magenta lines. The electron density for QW-5–70 is shown as a blue mesh; the final 2Fo-Fc map is contoured at 1.0 σ. Middle panel: detailed interactions of colchicine (light-brown sticks) binding to sT2R (PDB 6XER) for comparison. Right panel: superposition of sT2R complexes with QW-5–70 and colchicine sT2R (colored grey) complexes, highlighting the relative inhibitor binding positions and differing conformations of the α-T5 and β-T7 loops.

    Journal: Molecular cancer therapeutics

    Article Title: QW-5-70 targets the colchicine site and demonstrates antitumor activity in P-gp–overexpressing cancer models

    doi: 10.1158/1535-7163.MCT-25-1013

    Figure Lengend Snippet: (A) General Synthesis of QW-5–70. i). 60% NaH, SEMCl, THF, 0–25 °C; ii). n-BuLi, 3,4,5-trimethoxybenzaldehyde, THF, −78 °C; iii). Dess-Martin, CH 2 Cl 2 , r.t.; iv)1-(phenylsulfonyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole, Pd(PPh 3 ) 4 , Na 2 CO 3 , 1,4-dioxane/H 2 O (v/v = 2/1), reflux; v). Pd(OAC) 2 , PPh 3 , K 2 CO 3 , n-butanol, reflux; vi). 37% HCl, MeOH, reflux. (B) QW-5–70 induced tubulin depolymerization. Colchicine and paclitaxel were included as reference compounds. (C) SPR sensorgrams of QW-5–70 and colchicine binding to tubulin. (mean ± SEM, n = 3) (D) EBI competition assays. The upper β-tubulin band represents native β-tubulin, while the lower band corresponds to the EBI–β-tubulin adduct. GAPDH served as a loading control. (E) Immunoblot analysis of soluble and polymerized β-tubulin in PC-3 cells following treatment with QW-5–70, colchicine, or paclitaxel (20 nM). Cells were fractionated into soluble and polymerized tubulin pools, which were analyzed by immunoblotting. Ponceau S staining and GAPDH were used as loading controls for polymerized and soluble fractions, respectively. Quantification of polymerized β-tubulin expressed as a percentage of total β-tubulin (soluble + polymerized). (mean ± SEM, n=3). (F) Representative immunofluorescence images of interphase and mitotic SK-N-BE(2)-C, NB-1691, and PC-3 cells treated with QW-5–70 (2 nM and 5 nM) for 24 h. Microtubules were stained with anti-α-tubulin antibodies (red), and nuclei were counterstained with DAPI (blue). Scale bar = 50 μm (n=3). (G) High-resolution X-ray crystal structure of the sT2R complex with QW-5–70. Left panel: detailed interactions of QW-5–70 (green sticks) within the colchicine-binding site are shown. Bound water molecules are represented as red spheres, and hydrogen bonds are indicated by dashed magenta lines. The electron density for QW-5–70 is shown as a blue mesh; the final 2Fo-Fc map is contoured at 1.0 σ. Middle panel: detailed interactions of colchicine (light-brown sticks) binding to sT2R (PDB 6XER) for comparison. Right panel: superposition of sT2R complexes with QW-5–70 and colchicine sT2R (colored grey) complexes, highlighting the relative inhibitor binding positions and differing conformations of the α-T5 and β-T7 loops.

    Article Snippet: Human neuroblastoma (NB) cell lines (SK-N-BE(2)-C, RRID: CVCL_0529; NB-1691, RRID: CVCL_5628; SK-N-BE(2), RRID: CVCL_0528; SiMa, RRID: CVCL_1695; IMR-32, RRID: CVCL_0346) and prostate cancer cell lines (PC-3, RRID: CVCL_0035; 22Rv1, RRID: CVCL_1045) were originally obtained from ATCC in 2019.

    Techniques: Binding Assay, Reflux, Control, Western Blot, Staining, Immunofluorescence, Comparison

    (A) Representative images and quantification of colony formation in BE2C/VCR cells treated with QW-5–70, vincristine, or colchicine (1–5 nM). (B) Colony formation assays in PC-3/TxR cells treated with QW-5–70, paclitaxel, or colchicine (1–5 nM). Colony area is expressed as mean ± SEM relative to vehicle control (n = 5). (C) Transwell migration of BE2C/VCR (left) and PC-3/TxR (right) cells after 24 h treatment with vincristine or paclitaxel (5 nM) or QW-5–70 (2 or 5 nM). (D) Transwell migration of BE2C/VCR (left) and PC-3/TxR (right) cells treated with QW-5–70 (0.5–5 nM) for 24 h. Migration area is expressed as mean ± SEM relative to control (n = 5). Scale bar = 100 μm. (E) Immunoblot analysis of P-gp expression in SK-N-BE(2)-C and BE2C/VCR cells. (F) Viability of BE2C/VCR cells treated with QW-5–70 or vincristine (0.1 nM–3 μM) in the absence or presence of verapamil (10 μM) for 72 h (MTS assay; n = 4). (G) Intracellular concentrations of QW-5–70 and vincristine in parental SK-N-BE(2)-C and BE2C/VCR cells following 50 nM, 2 h treatment, with or without tariquidar (1 μM), quantified by LC–MS/MS (n = 3). (H) Immunoblot analysis of P-gp expression in PC-3 and PC-3/TxR cells. (I) Cell viability of PC-3/TxR cells treated with QW-5–70 or paclitaxel (0.1 nM–3 μM) in the absence or presence of verapamil (10 μM) for 72 h (MTS assay; n = 4). (J) Intracellular concentrations of QW-5–70 and paclitaxel in PC-3 and PC-3/TxR cells under the same conditions as panel G, quantified by LC–MS/MS (n = 3). (K) Immunoblot analysis of P-gp expression in PC-3/TxR cells following transfection with scrambled control or si-P-gp. GAPDH served as a loading control. (L) Viability of QW-5–70 and paclitaxel in PC-3/TxR, PC-3/TxR-scramble, and PC-3/TxR-si-P-gp cells (MTS assay; n=4).

    Journal: Molecular cancer therapeutics

    Article Title: QW-5-70 targets the colchicine site and demonstrates antitumor activity in P-gp–overexpressing cancer models

    doi: 10.1158/1535-7163.MCT-25-1013

    Figure Lengend Snippet: (A) Representative images and quantification of colony formation in BE2C/VCR cells treated with QW-5–70, vincristine, or colchicine (1–5 nM). (B) Colony formation assays in PC-3/TxR cells treated with QW-5–70, paclitaxel, or colchicine (1–5 nM). Colony area is expressed as mean ± SEM relative to vehicle control (n = 5). (C) Transwell migration of BE2C/VCR (left) and PC-3/TxR (right) cells after 24 h treatment with vincristine or paclitaxel (5 nM) or QW-5–70 (2 or 5 nM). (D) Transwell migration of BE2C/VCR (left) and PC-3/TxR (right) cells treated with QW-5–70 (0.5–5 nM) for 24 h. Migration area is expressed as mean ± SEM relative to control (n = 5). Scale bar = 100 μm. (E) Immunoblot analysis of P-gp expression in SK-N-BE(2)-C and BE2C/VCR cells. (F) Viability of BE2C/VCR cells treated with QW-5–70 or vincristine (0.1 nM–3 μM) in the absence or presence of verapamil (10 μM) for 72 h (MTS assay; n = 4). (G) Intracellular concentrations of QW-5–70 and vincristine in parental SK-N-BE(2)-C and BE2C/VCR cells following 50 nM, 2 h treatment, with or without tariquidar (1 μM), quantified by LC–MS/MS (n = 3). (H) Immunoblot analysis of P-gp expression in PC-3 and PC-3/TxR cells. (I) Cell viability of PC-3/TxR cells treated with QW-5–70 or paclitaxel (0.1 nM–3 μM) in the absence or presence of verapamil (10 μM) for 72 h (MTS assay; n = 4). (J) Intracellular concentrations of QW-5–70 and paclitaxel in PC-3 and PC-3/TxR cells under the same conditions as panel G, quantified by LC–MS/MS (n = 3). (K) Immunoblot analysis of P-gp expression in PC-3/TxR cells following transfection with scrambled control or si-P-gp. GAPDH served as a loading control. (L) Viability of QW-5–70 and paclitaxel in PC-3/TxR, PC-3/TxR-scramble, and PC-3/TxR-si-P-gp cells (MTS assay; n=4).

    Article Snippet: Human neuroblastoma (NB) cell lines (SK-N-BE(2)-C, RRID: CVCL_0529; NB-1691, RRID: CVCL_5628; SK-N-BE(2), RRID: CVCL_0528; SiMa, RRID: CVCL_1695; IMR-32, RRID: CVCL_0346) and prostate cancer cell lines (PC-3, RRID: CVCL_0035; 22Rv1, RRID: CVCL_1045) were originally obtained from ATCC in 2019.

    Techniques: Activity Assay, Expressing, Control, Migration, Western Blot, MTS Assay, Liquid Chromatography with Mass Spectroscopy, Transfection

    (A) Cell-cycle distribution of SK-N-BE(2)-C, NB-1691, and PC-3/TxR cells treated with QW-5–70 (2 or 5 nM) for 24 h, analyzed by flow cytometry (n=3). (B, C) Immunoblot analysis of cell-cycle–related proteins (phospho-histone H3 (Ser10), histone H3, cyclin B1, phospho-CDK1 (Thr161), and CDK1) in parental (SK-N-BE(2)-C, NB-1691) and resistant (BE2C/VCR, PC-3/TxR) cells treated with QW-5–70 (2 or 5 nM) for 24 h. (D, E) Immunoblot analysis of apoptosis-related proteins (cCas9, cCas3, PARP, cPARP, p-BCL2 (Ser70), and BCL2) in the same cell lines following 24 h treatment. GAPDH served as a loading control.

    Journal: Molecular cancer therapeutics

    Article Title: QW-5-70 targets the colchicine site and demonstrates antitumor activity in P-gp–overexpressing cancer models

    doi: 10.1158/1535-7163.MCT-25-1013

    Figure Lengend Snippet: (A) Cell-cycle distribution of SK-N-BE(2)-C, NB-1691, and PC-3/TxR cells treated with QW-5–70 (2 or 5 nM) for 24 h, analyzed by flow cytometry (n=3). (B, C) Immunoblot analysis of cell-cycle–related proteins (phospho-histone H3 (Ser10), histone H3, cyclin B1, phospho-CDK1 (Thr161), and CDK1) in parental (SK-N-BE(2)-C, NB-1691) and resistant (BE2C/VCR, PC-3/TxR) cells treated with QW-5–70 (2 or 5 nM) for 24 h. (D, E) Immunoblot analysis of apoptosis-related proteins (cCas9, cCas3, PARP, cPARP, p-BCL2 (Ser70), and BCL2) in the same cell lines following 24 h treatment. GAPDH served as a loading control.

    Article Snippet: Human neuroblastoma (NB) cell lines (SK-N-BE(2)-C, RRID: CVCL_0529; NB-1691, RRID: CVCL_5628; SK-N-BE(2), RRID: CVCL_0528; SiMa, RRID: CVCL_1695; IMR-32, RRID: CVCL_0346) and prostate cancer cell lines (PC-3, RRID: CVCL_0035; 22Rv1, RRID: CVCL_1045) were originally obtained from ATCC in 2019.

    Techniques: Flow Cytometry, Western Blot, Control

    (A) SK-N-BE(2)-c cells were treated with five doses of QW-5–70 (0.125 to 2 nM) in combination with five doses of DFMO (10–1000 μM). Cell viability was assessed using the MTS assay (n=4). (B) CI–Fa analysis of the QW-5–70 and DFMO combination. CI values were calculated using the Chou–Talalay median-effect method for five experimentally tested dose pairs at their corresponding Fa values. CI < 1, CI = 1, and CI > 1 indicate synergistic, additive, and antagonistic interactions, respectively. (C) SK-N-BE(2)-c cells were treated with five doses of QW-5–70 (0.125 to 2 nM) in combination with five doses of MLN8237 (1–100 nM). Cell viability was assessed using the MTS assay (n=4). (D) SK-N-BE(2)-C cells were treated with 1 nM QW-5–70 in combination with either DFMO (300 μM, top) or MLN8237 (30 nM, bottom) as indicated. Cell lysates were analyzed by immunoblotting for cPARP as a marker of apoptosis, with GAPDH used as a loading control. (E) Representative colony formation images of SK-N-BE(2)-C cells treated with the same combination. Colony formations were quantified and expressed as mean ± SEM relative to vehicle (set to 100%) (n=5).

    Journal: Molecular cancer therapeutics

    Article Title: QW-5-70 targets the colchicine site and demonstrates antitumor activity in P-gp–overexpressing cancer models

    doi: 10.1158/1535-7163.MCT-25-1013

    Figure Lengend Snippet: (A) SK-N-BE(2)-c cells were treated with five doses of QW-5–70 (0.125 to 2 nM) in combination with five doses of DFMO (10–1000 μM). Cell viability was assessed using the MTS assay (n=4). (B) CI–Fa analysis of the QW-5–70 and DFMO combination. CI values were calculated using the Chou–Talalay median-effect method for five experimentally tested dose pairs at their corresponding Fa values. CI < 1, CI = 1, and CI > 1 indicate synergistic, additive, and antagonistic interactions, respectively. (C) SK-N-BE(2)-c cells were treated with five doses of QW-5–70 (0.125 to 2 nM) in combination with five doses of MLN8237 (1–100 nM). Cell viability was assessed using the MTS assay (n=4). (D) SK-N-BE(2)-C cells were treated with 1 nM QW-5–70 in combination with either DFMO (300 μM, top) or MLN8237 (30 nM, bottom) as indicated. Cell lysates were analyzed by immunoblotting for cPARP as a marker of apoptosis, with GAPDH used as a loading control. (E) Representative colony formation images of SK-N-BE(2)-C cells treated with the same combination. Colony formations were quantified and expressed as mean ± SEM relative to vehicle (set to 100%) (n=5).

    Article Snippet: Human neuroblastoma (NB) cell lines (SK-N-BE(2)-C, RRID: CVCL_0529; NB-1691, RRID: CVCL_5628; SK-N-BE(2), RRID: CVCL_0528; SiMa, RRID: CVCL_1695; IMR-32, RRID: CVCL_0346) and prostate cancer cell lines (PC-3, RRID: CVCL_0035; 22Rv1, RRID: CVCL_1045) were originally obtained from ATCC in 2019.

    Techniques: Activity Assay, MTS Assay, Western Blot, Marker, Control