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primary antibodies against total egfr  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc primary antibodies against total egfr
    Effects of Ncs on apoptosis and cell cycle. ( A ) Cells were treated with 50 nM Ncs for 24 h and stained with calcein-AM/PI, followed by image capture using fluorescence microscopy (green: live cells, red; dead cells), scale bars = 50 μm. ( B ) cells were treated with 50 nM Ncs for 24 h and stained with annexin V and PI and followed by flow cytometric analysis of apoptosis. Apoptotic cells were quantified with early and late apoptotic cell populations. ( C ) For cell cycle arrays, 50 nM Ncs-treated cells were fixed with 70% ethanol and stained with PI, followed by flow cytometry analysis. ( D and E ) Cells were treated with indicated concentrations (0–100 nM) of Ncs for 24 h and cell lysates were subjected to immunoblotting analysis using the <t>indicated</t> <t>antibodies.</t> Expression levels of <t>p-EGFR</t> and p-ERK1/2 were quantified from immunoblot bands and normalized to total EGFR and total ERK1/2 expression levels, respectively ( D ). Data were presented as mean values from three independent experiments and error bars represent standard deviations. * p < 0.05, ** p < 0.01
    Primary Antibodies Against Total Egfr, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1533 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/rabbit+anti+total+egfr/pmc12679757-55-0-6?v=Cell+Signaling+Technology+Inc
    Average 96 stars, based on 1533 article reviews
    primary antibodies against total egfr - by Bioz Stars, 2026-08
    96/100 stars

    Images

    1) Product Images from "Narciclasine as a potential therapeutic agent to overcome EGFR-TKI resistance in non-small cell lung cancer"

    Article Title: Narciclasine as a potential therapeutic agent to overcome EGFR-TKI resistance in non-small cell lung cancer

    Journal: Journal of Translational Medicine

    doi: 10.1186/s12967-025-07368-4

    Effects of Ncs on apoptosis and cell cycle. ( A ) Cells were treated with 50 nM Ncs for 24 h and stained with calcein-AM/PI, followed by image capture using fluorescence microscopy (green: live cells, red; dead cells), scale bars = 50 μm. ( B ) cells were treated with 50 nM Ncs for 24 h and stained with annexin V and PI and followed by flow cytometric analysis of apoptosis. Apoptotic cells were quantified with early and late apoptotic cell populations. ( C ) For cell cycle arrays, 50 nM Ncs-treated cells were fixed with 70% ethanol and stained with PI, followed by flow cytometry analysis. ( D and E ) Cells were treated with indicated concentrations (0–100 nM) of Ncs for 24 h and cell lysates were subjected to immunoblotting analysis using the indicated antibodies. Expression levels of p-EGFR and p-ERK1/2 were quantified from immunoblot bands and normalized to total EGFR and total ERK1/2 expression levels, respectively ( D ). Data were presented as mean values from three independent experiments and error bars represent standard deviations. * p < 0.05, ** p < 0.01
    Figure Legend Snippet: Effects of Ncs on apoptosis and cell cycle. ( A ) Cells were treated with 50 nM Ncs for 24 h and stained with calcein-AM/PI, followed by image capture using fluorescence microscopy (green: live cells, red; dead cells), scale bars = 50 μm. ( B ) cells were treated with 50 nM Ncs for 24 h and stained with annexin V and PI and followed by flow cytometric analysis of apoptosis. Apoptotic cells were quantified with early and late apoptotic cell populations. ( C ) For cell cycle arrays, 50 nM Ncs-treated cells were fixed with 70% ethanol and stained with PI, followed by flow cytometry analysis. ( D and E ) Cells were treated with indicated concentrations (0–100 nM) of Ncs for 24 h and cell lysates were subjected to immunoblotting analysis using the indicated antibodies. Expression levels of p-EGFR and p-ERK1/2 were quantified from immunoblot bands and normalized to total EGFR and total ERK1/2 expression levels, respectively ( D ). Data were presented as mean values from three independent experiments and error bars represent standard deviations. * p < 0.05, ** p < 0.01

    Techniques Used: Staining, Fluorescence, Microscopy, Flow Cytometry, Western Blot, Expressing

    Effects of Ncs on cell growth in PC-9 and PC-9-GR cells. ( A ) PC-9 and PC-9-GR cells (3 × 10 3 cells/well) were plated in 96-well plates and treated with gefitinib in a dose-dependent manner for 48 h. ( B ) Cells were treated with Ncs at the indicated concentrations for 48 hours, and cell growth rates were measured using the WST-1 assay. ( C ) Cells were treated with Ncs for 48 hours, and representative images are shown. ( D ) Cells (1 × 10 4 cells/well) were plated in 24-well plates and treated with Ncs at the indicated concentrations for 48 h. Live and dead cells were counted using the trypan blue staining assay. ( E and F ) Cells were treated with Ncs for 48 h, fixed with 70% ethanol, stained with PI, and analyzed by flow cytometry to assess cell cycle distribution. ( G ) PC-9 and PC-9-GR cells were treated with the indicated concentrations (0-50 nM) of Ncs for 24 h. Cell lysates were then subjected to immunoblotting analysis using the indicated antibodies. Expression levels of p-EGFR, p-Src, and p-STAT3 were quantified and normalized to total EGFR, total Src, and total STAT3 expression levels, respectively. Data were presented as mean values from three independent experiments and error bars represent standard deviations of the mean of three measurements. Magnification: x 100, n.s. >0.05, *P < 0.05, **P < 0.01
    Figure Legend Snippet: Effects of Ncs on cell growth in PC-9 and PC-9-GR cells. ( A ) PC-9 and PC-9-GR cells (3 × 10 3 cells/well) were plated in 96-well plates and treated with gefitinib in a dose-dependent manner for 48 h. ( B ) Cells were treated with Ncs at the indicated concentrations for 48 hours, and cell growth rates were measured using the WST-1 assay. ( C ) Cells were treated with Ncs for 48 hours, and representative images are shown. ( D ) Cells (1 × 10 4 cells/well) were plated in 24-well plates and treated with Ncs at the indicated concentrations for 48 h. Live and dead cells were counted using the trypan blue staining assay. ( E and F ) Cells were treated with Ncs for 48 h, fixed with 70% ethanol, stained with PI, and analyzed by flow cytometry to assess cell cycle distribution. ( G ) PC-9 and PC-9-GR cells were treated with the indicated concentrations (0-50 nM) of Ncs for 24 h. Cell lysates were then subjected to immunoblotting analysis using the indicated antibodies. Expression levels of p-EGFR, p-Src, and p-STAT3 were quantified and normalized to total EGFR, total Src, and total STAT3 expression levels, respectively. Data were presented as mean values from three independent experiments and error bars represent standard deviations of the mean of three measurements. Magnification: x 100, n.s. >0.05, *P < 0.05, **P < 0.01

    Techniques Used: WST-1 Assay, Staining, Flow Cytometry, Western Blot, Expressing

    Effects of Ncs on EGFR activity and EGFR signaling. ( A ) H1975 cells were treated with 50 nM of Ncs for indicated times and cell lysates were subjected to immunoblotting analysis using the indicated antibodies. ( B ) Cells were treated with 50 nM of Ncs for the indicated durations, and total RNA was isolated. EGFR mRNA levels were determined by real-time PCR analysis and normalized to GAPDH expression. ( C ) Cells were treated with 20 μM of cycloheximide (CHX) for the indicated time points in the presence of either DMSO or 50 nM of Ncs for 24 h. Cell lysates were subjected to immunoblotting analysis using the indicated antibodies (upper panel). EGFR expression levels were quantified from immunoblot bands and normalized to GAPDH, comparing CHX treatment in the presence of DMSO or Ncs (lower panel). ( D ) Cells were treated with 50 nM of Ncs for 24 h in the presence of BafA1 (20 nM, 12 h) or MG132 (20 μM, 2 h). Cell lysates were subjected to immunoblotting analysis using indicated antibodies. ( E ) Cells were plated on coverslips and treated with DMSO (control) or 50 nM of Ncs for 24 h. Cells were then fixed with 4% paraformaldehyde (PFA) and subjected to immunofluorescence staining using DAPI, EGFR, and LAMP1 antibody. ( F ) Cells were treated with 50 nM of Ncs for 24 h, followed by EGF (10 nM, 15 min) stimulation. Cell lysates were subjected to immunoblotting analysis using the specified antibodies. Similar results were observed in three independent experiments. Error bars represent standard deviations of the mean of three measurements, *P < 0.05, **P < 0.01, ***P < 0.001
    Figure Legend Snippet: Effects of Ncs on EGFR activity and EGFR signaling. ( A ) H1975 cells were treated with 50 nM of Ncs for indicated times and cell lysates were subjected to immunoblotting analysis using the indicated antibodies. ( B ) Cells were treated with 50 nM of Ncs for the indicated durations, and total RNA was isolated. EGFR mRNA levels were determined by real-time PCR analysis and normalized to GAPDH expression. ( C ) Cells were treated with 20 μM of cycloheximide (CHX) for the indicated time points in the presence of either DMSO or 50 nM of Ncs for 24 h. Cell lysates were subjected to immunoblotting analysis using the indicated antibodies (upper panel). EGFR expression levels were quantified from immunoblot bands and normalized to GAPDH, comparing CHX treatment in the presence of DMSO or Ncs (lower panel). ( D ) Cells were treated with 50 nM of Ncs for 24 h in the presence of BafA1 (20 nM, 12 h) or MG132 (20 μM, 2 h). Cell lysates were subjected to immunoblotting analysis using indicated antibodies. ( E ) Cells were plated on coverslips and treated with DMSO (control) or 50 nM of Ncs for 24 h. Cells were then fixed with 4% paraformaldehyde (PFA) and subjected to immunofluorescence staining using DAPI, EGFR, and LAMP1 antibody. ( F ) Cells were treated with 50 nM of Ncs for 24 h, followed by EGF (10 nM, 15 min) stimulation. Cell lysates were subjected to immunoblotting analysis using the specified antibodies. Similar results were observed in three independent experiments. Error bars represent standard deviations of the mean of three measurements, *P < 0.05, **P < 0.01, ***P < 0.001

    Techniques Used: Activity Assay, Western Blot, Isolation, Real-time Polymerase Chain Reaction, Expressing, Control, Immunofluorescence, Staining

    Anti-cancer effects of ncs in vivo. ( A ) Adult-stage C. elegans models: wild type (WT) and jgIs25 strains. The jgIs25 strain exhibits polyps on the ventral side (arrows in the lower panel). Scale bar = 20 μm. ( B ) Effects of gefitinib and ncs treatment on jgIs25 . the arrows in the upper panel indicate polyps, and the rectangular area is magnified in the lower panel. Scale bar = 200 μm (upper), 20 μm (lower). ( C ) Quantification of polyp formation in jgIs25 following treatment with the indicated drugs. ( D and G ) In vivo tumor model using subcutaneous transplantation of A549 and H1975 cells into nude mice, followed by ncs treatment. Tumor volumes were measured at the indicated time points in control and ncs-treated mice. Tumor weights were measured after sacrifice. ( E and H ) Representative image of xenograft tumors after sacrifice. ( F and I ) Immunohistochemical (IHC) staining of xenograft tumors using anti-EGFR, anti-phospho-EGFR (Y1068), and anti-phospho-STAT3 (Y705) antibodies. Scale bar = 500 μm. Similar results were observed in three independent experiments. Error bars represent standard deviations of the mean of three measurements, *p < 0.05, **p < 0.01, *p < 0.001
    Figure Legend Snippet: Anti-cancer effects of ncs in vivo. ( A ) Adult-stage C. elegans models: wild type (WT) and jgIs25 strains. The jgIs25 strain exhibits polyps on the ventral side (arrows in the lower panel). Scale bar = 20 μm. ( B ) Effects of gefitinib and ncs treatment on jgIs25 . the arrows in the upper panel indicate polyps, and the rectangular area is magnified in the lower panel. Scale bar = 200 μm (upper), 20 μm (lower). ( C ) Quantification of polyp formation in jgIs25 following treatment with the indicated drugs. ( D and G ) In vivo tumor model using subcutaneous transplantation of A549 and H1975 cells into nude mice, followed by ncs treatment. Tumor volumes were measured at the indicated time points in control and ncs-treated mice. Tumor weights were measured after sacrifice. ( E and H ) Representative image of xenograft tumors after sacrifice. ( F and I ) Immunohistochemical (IHC) staining of xenograft tumors using anti-EGFR, anti-phospho-EGFR (Y1068), and anti-phospho-STAT3 (Y705) antibodies. Scale bar = 500 μm. Similar results were observed in three independent experiments. Error bars represent standard deviations of the mean of three measurements, *p < 0.05, **p < 0.01, *p < 0.001

    Techniques Used: In Vivo, Transplantation Assay, Control, Immunohistochemical staining, Immunohistochemistry



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    Effects of Ncs on apoptosis and cell cycle. ( A ) Cells were treated with 50 nM Ncs for 24 h and stained with calcein-AM/PI, followed by image capture using fluorescence microscopy (green: live cells, red; dead cells), scale bars = 50 μm. ( B ) cells were treated with 50 nM Ncs for 24 h and stained with annexin V and PI and followed by flow cytometric analysis of apoptosis. Apoptotic cells were quantified with early and late apoptotic cell populations. ( C ) For cell cycle arrays, 50 nM Ncs-treated cells were fixed with 70% ethanol and stained with PI, followed by flow cytometry analysis. ( D and E ) Cells were treated with indicated concentrations (0–100 nM) of Ncs for 24 h and cell lysates were subjected to immunoblotting analysis using the <t>indicated</t> <t>antibodies.</t> Expression levels of <t>p-EGFR</t> and p-ERK1/2 were quantified from immunoblot bands and normalized to total EGFR and total ERK1/2 expression levels, respectively ( D ). Data were presented as mean values from three independent experiments and error bars represent standard deviations. * p < 0.05, ** p < 0.01
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    Effects of Ncs on apoptosis and cell cycle. ( A ) Cells were treated with 50 nM Ncs for 24 h and stained with calcein-AM/PI, followed by image capture using fluorescence microscopy (green: live cells, red; dead cells), scale bars = 50 μm. ( B ) cells were treated with 50 nM Ncs for 24 h and stained with annexin V and PI and followed by flow cytometric analysis of apoptosis. Apoptotic cells were quantified with early and late apoptotic cell populations. ( C ) For cell cycle arrays, 50 nM Ncs-treated cells were fixed with 70% ethanol and stained with PI, followed by flow cytometry analysis. ( D and E ) Cells were treated with indicated concentrations (0–100 nM) of Ncs for 24 h and cell lysates were subjected to immunoblotting analysis using the indicated antibodies. Expression levels of p-EGFR and p-ERK1/2 were quantified from immunoblot bands and normalized to total EGFR and total ERK1/2 expression levels, respectively ( D ). Data were presented as mean values from three independent experiments and error bars represent standard deviations. * p < 0.05, ** p < 0.01

    Journal: Journal of Translational Medicine

    Article Title: Narciclasine as a potential therapeutic agent to overcome EGFR-TKI resistance in non-small cell lung cancer

    doi: 10.1186/s12967-025-07368-4

    Figure Lengend Snippet: Effects of Ncs on apoptosis and cell cycle. ( A ) Cells were treated with 50 nM Ncs for 24 h and stained with calcein-AM/PI, followed by image capture using fluorescence microscopy (green: live cells, red; dead cells), scale bars = 50 μm. ( B ) cells were treated with 50 nM Ncs for 24 h and stained with annexin V and PI and followed by flow cytometric analysis of apoptosis. Apoptotic cells were quantified with early and late apoptotic cell populations. ( C ) For cell cycle arrays, 50 nM Ncs-treated cells were fixed with 70% ethanol and stained with PI, followed by flow cytometry analysis. ( D and E ) Cells were treated with indicated concentrations (0–100 nM) of Ncs for 24 h and cell lysates were subjected to immunoblotting analysis using the indicated antibodies. Expression levels of p-EGFR and p-ERK1/2 were quantified from immunoblot bands and normalized to total EGFR and total ERK1/2 expression levels, respectively ( D ). Data were presented as mean values from three independent experiments and error bars represent standard deviations. * p < 0.05, ** p < 0.01

    Article Snippet: Primary antibodies against total EGFR (#4267, Cell Signaling Technology(CST)), CDK1 (#9116, CST), phospho-EGFR Y1068 (#44-788 G, Invitrogen), total STAT3 (#4904, CST), phospho-STAT3 Y705 (#9145, CST), phospho-Src (#6943,CST), total-Src (#2109, CST), phospho-CDK1 Y15 (#4539, CST), phospho-Wee1 (#4910, CST), Myt1 (#4282, CST), p21 (#2947, CST), Cdc25B (#9525, CST), GAPDH (#5174, CST), β-actin (#A700-057, Bethyl Laboratory), phospho-ERK1/2 (#9101, CST), Bcl-2 (#sc-7382, Santa Cruz), and c-Myc (#sc-789, Santa Cruz) were used for immunoblotting.

    Techniques: Staining, Fluorescence, Microscopy, Flow Cytometry, Western Blot, Expressing

    Effects of Ncs on cell growth in PC-9 and PC-9-GR cells. ( A ) PC-9 and PC-9-GR cells (3 × 10 3 cells/well) were plated in 96-well plates and treated with gefitinib in a dose-dependent manner for 48 h. ( B ) Cells were treated with Ncs at the indicated concentrations for 48 hours, and cell growth rates were measured using the WST-1 assay. ( C ) Cells were treated with Ncs for 48 hours, and representative images are shown. ( D ) Cells (1 × 10 4 cells/well) were plated in 24-well plates and treated with Ncs at the indicated concentrations for 48 h. Live and dead cells were counted using the trypan blue staining assay. ( E and F ) Cells were treated with Ncs for 48 h, fixed with 70% ethanol, stained with PI, and analyzed by flow cytometry to assess cell cycle distribution. ( G ) PC-9 and PC-9-GR cells were treated with the indicated concentrations (0-50 nM) of Ncs for 24 h. Cell lysates were then subjected to immunoblotting analysis using the indicated antibodies. Expression levels of p-EGFR, p-Src, and p-STAT3 were quantified and normalized to total EGFR, total Src, and total STAT3 expression levels, respectively. Data were presented as mean values from three independent experiments and error bars represent standard deviations of the mean of three measurements. Magnification: x 100, n.s. >0.05, *P < 0.05, **P < 0.01

    Journal: Journal of Translational Medicine

    Article Title: Narciclasine as a potential therapeutic agent to overcome EGFR-TKI resistance in non-small cell lung cancer

    doi: 10.1186/s12967-025-07368-4

    Figure Lengend Snippet: Effects of Ncs on cell growth in PC-9 and PC-9-GR cells. ( A ) PC-9 and PC-9-GR cells (3 × 10 3 cells/well) were plated in 96-well plates and treated with gefitinib in a dose-dependent manner for 48 h. ( B ) Cells were treated with Ncs at the indicated concentrations for 48 hours, and cell growth rates were measured using the WST-1 assay. ( C ) Cells were treated with Ncs for 48 hours, and representative images are shown. ( D ) Cells (1 × 10 4 cells/well) were plated in 24-well plates and treated with Ncs at the indicated concentrations for 48 h. Live and dead cells were counted using the trypan blue staining assay. ( E and F ) Cells were treated with Ncs for 48 h, fixed with 70% ethanol, stained with PI, and analyzed by flow cytometry to assess cell cycle distribution. ( G ) PC-9 and PC-9-GR cells were treated with the indicated concentrations (0-50 nM) of Ncs for 24 h. Cell lysates were then subjected to immunoblotting analysis using the indicated antibodies. Expression levels of p-EGFR, p-Src, and p-STAT3 were quantified and normalized to total EGFR, total Src, and total STAT3 expression levels, respectively. Data were presented as mean values from three independent experiments and error bars represent standard deviations of the mean of three measurements. Magnification: x 100, n.s. >0.05, *P < 0.05, **P < 0.01

    Article Snippet: Primary antibodies against total EGFR (#4267, Cell Signaling Technology(CST)), CDK1 (#9116, CST), phospho-EGFR Y1068 (#44-788 G, Invitrogen), total STAT3 (#4904, CST), phospho-STAT3 Y705 (#9145, CST), phospho-Src (#6943,CST), total-Src (#2109, CST), phospho-CDK1 Y15 (#4539, CST), phospho-Wee1 (#4910, CST), Myt1 (#4282, CST), p21 (#2947, CST), Cdc25B (#9525, CST), GAPDH (#5174, CST), β-actin (#A700-057, Bethyl Laboratory), phospho-ERK1/2 (#9101, CST), Bcl-2 (#sc-7382, Santa Cruz), and c-Myc (#sc-789, Santa Cruz) were used for immunoblotting.

    Techniques: WST-1 Assay, Staining, Flow Cytometry, Western Blot, Expressing

    Effects of Ncs on EGFR activity and EGFR signaling. ( A ) H1975 cells were treated with 50 nM of Ncs for indicated times and cell lysates were subjected to immunoblotting analysis using the indicated antibodies. ( B ) Cells were treated with 50 nM of Ncs for the indicated durations, and total RNA was isolated. EGFR mRNA levels were determined by real-time PCR analysis and normalized to GAPDH expression. ( C ) Cells were treated with 20 μM of cycloheximide (CHX) for the indicated time points in the presence of either DMSO or 50 nM of Ncs for 24 h. Cell lysates were subjected to immunoblotting analysis using the indicated antibodies (upper panel). EGFR expression levels were quantified from immunoblot bands and normalized to GAPDH, comparing CHX treatment in the presence of DMSO or Ncs (lower panel). ( D ) Cells were treated with 50 nM of Ncs for 24 h in the presence of BafA1 (20 nM, 12 h) or MG132 (20 μM, 2 h). Cell lysates were subjected to immunoblotting analysis using indicated antibodies. ( E ) Cells were plated on coverslips and treated with DMSO (control) or 50 nM of Ncs for 24 h. Cells were then fixed with 4% paraformaldehyde (PFA) and subjected to immunofluorescence staining using DAPI, EGFR, and LAMP1 antibody. ( F ) Cells were treated with 50 nM of Ncs for 24 h, followed by EGF (10 nM, 15 min) stimulation. Cell lysates were subjected to immunoblotting analysis using the specified antibodies. Similar results were observed in three independent experiments. Error bars represent standard deviations of the mean of three measurements, *P < 0.05, **P < 0.01, ***P < 0.001

    Journal: Journal of Translational Medicine

    Article Title: Narciclasine as a potential therapeutic agent to overcome EGFR-TKI resistance in non-small cell lung cancer

    doi: 10.1186/s12967-025-07368-4

    Figure Lengend Snippet: Effects of Ncs on EGFR activity and EGFR signaling. ( A ) H1975 cells were treated with 50 nM of Ncs for indicated times and cell lysates were subjected to immunoblotting analysis using the indicated antibodies. ( B ) Cells were treated with 50 nM of Ncs for the indicated durations, and total RNA was isolated. EGFR mRNA levels were determined by real-time PCR analysis and normalized to GAPDH expression. ( C ) Cells were treated with 20 μM of cycloheximide (CHX) for the indicated time points in the presence of either DMSO or 50 nM of Ncs for 24 h. Cell lysates were subjected to immunoblotting analysis using the indicated antibodies (upper panel). EGFR expression levels were quantified from immunoblot bands and normalized to GAPDH, comparing CHX treatment in the presence of DMSO or Ncs (lower panel). ( D ) Cells were treated with 50 nM of Ncs for 24 h in the presence of BafA1 (20 nM, 12 h) or MG132 (20 μM, 2 h). Cell lysates were subjected to immunoblotting analysis using indicated antibodies. ( E ) Cells were plated on coverslips and treated with DMSO (control) or 50 nM of Ncs for 24 h. Cells were then fixed with 4% paraformaldehyde (PFA) and subjected to immunofluorescence staining using DAPI, EGFR, and LAMP1 antibody. ( F ) Cells were treated with 50 nM of Ncs for 24 h, followed by EGF (10 nM, 15 min) stimulation. Cell lysates were subjected to immunoblotting analysis using the specified antibodies. Similar results were observed in three independent experiments. Error bars represent standard deviations of the mean of three measurements, *P < 0.05, **P < 0.01, ***P < 0.001

    Article Snippet: Primary antibodies against total EGFR (#4267, Cell Signaling Technology(CST)), CDK1 (#9116, CST), phospho-EGFR Y1068 (#44-788 G, Invitrogen), total STAT3 (#4904, CST), phospho-STAT3 Y705 (#9145, CST), phospho-Src (#6943,CST), total-Src (#2109, CST), phospho-CDK1 Y15 (#4539, CST), phospho-Wee1 (#4910, CST), Myt1 (#4282, CST), p21 (#2947, CST), Cdc25B (#9525, CST), GAPDH (#5174, CST), β-actin (#A700-057, Bethyl Laboratory), phospho-ERK1/2 (#9101, CST), Bcl-2 (#sc-7382, Santa Cruz), and c-Myc (#sc-789, Santa Cruz) were used for immunoblotting.

    Techniques: Activity Assay, Western Blot, Isolation, Real-time Polymerase Chain Reaction, Expressing, Control, Immunofluorescence, Staining

    Anti-cancer effects of ncs in vivo. ( A ) Adult-stage C. elegans models: wild type (WT) and jgIs25 strains. The jgIs25 strain exhibits polyps on the ventral side (arrows in the lower panel). Scale bar = 20 μm. ( B ) Effects of gefitinib and ncs treatment on jgIs25 . the arrows in the upper panel indicate polyps, and the rectangular area is magnified in the lower panel. Scale bar = 200 μm (upper), 20 μm (lower). ( C ) Quantification of polyp formation in jgIs25 following treatment with the indicated drugs. ( D and G ) In vivo tumor model using subcutaneous transplantation of A549 and H1975 cells into nude mice, followed by ncs treatment. Tumor volumes were measured at the indicated time points in control and ncs-treated mice. Tumor weights were measured after sacrifice. ( E and H ) Representative image of xenograft tumors after sacrifice. ( F and I ) Immunohistochemical (IHC) staining of xenograft tumors using anti-EGFR, anti-phospho-EGFR (Y1068), and anti-phospho-STAT3 (Y705) antibodies. Scale bar = 500 μm. Similar results were observed in three independent experiments. Error bars represent standard deviations of the mean of three measurements, *p < 0.05, **p < 0.01, *p < 0.001

    Journal: Journal of Translational Medicine

    Article Title: Narciclasine as a potential therapeutic agent to overcome EGFR-TKI resistance in non-small cell lung cancer

    doi: 10.1186/s12967-025-07368-4

    Figure Lengend Snippet: Anti-cancer effects of ncs in vivo. ( A ) Adult-stage C. elegans models: wild type (WT) and jgIs25 strains. The jgIs25 strain exhibits polyps on the ventral side (arrows in the lower panel). Scale bar = 20 μm. ( B ) Effects of gefitinib and ncs treatment on jgIs25 . the arrows in the upper panel indicate polyps, and the rectangular area is magnified in the lower panel. Scale bar = 200 μm (upper), 20 μm (lower). ( C ) Quantification of polyp formation in jgIs25 following treatment with the indicated drugs. ( D and G ) In vivo tumor model using subcutaneous transplantation of A549 and H1975 cells into nude mice, followed by ncs treatment. Tumor volumes were measured at the indicated time points in control and ncs-treated mice. Tumor weights were measured after sacrifice. ( E and H ) Representative image of xenograft tumors after sacrifice. ( F and I ) Immunohistochemical (IHC) staining of xenograft tumors using anti-EGFR, anti-phospho-EGFR (Y1068), and anti-phospho-STAT3 (Y705) antibodies. Scale bar = 500 μm. Similar results were observed in three independent experiments. Error bars represent standard deviations of the mean of three measurements, *p < 0.05, **p < 0.01, *p < 0.001

    Article Snippet: Primary antibodies against total EGFR (#4267, Cell Signaling Technology(CST)), CDK1 (#9116, CST), phospho-EGFR Y1068 (#44-788 G, Invitrogen), total STAT3 (#4904, CST), phospho-STAT3 Y705 (#9145, CST), phospho-Src (#6943,CST), total-Src (#2109, CST), phospho-CDK1 Y15 (#4539, CST), phospho-Wee1 (#4910, CST), Myt1 (#4282, CST), p21 (#2947, CST), Cdc25B (#9525, CST), GAPDH (#5174, CST), β-actin (#A700-057, Bethyl Laboratory), phospho-ERK1/2 (#9101, CST), Bcl-2 (#sc-7382, Santa Cruz), and c-Myc (#sc-789, Santa Cruz) were used for immunoblotting.

    Techniques: In Vivo, Transplantation Assay, Control, Immunohistochemical staining, Immunohistochemistry

    a Summary of ELCOL. Treatment course for patients with clinical resistance to EGFR-TKI is shown on the right. Circle indicates the timing of tissue sampling and organoid derivation and is colored according to the histology. Thick line shows the treatment period. LUAD; lung adenocarcinoma, SQ-T; squamous transformation, SCLC-T; small cell lung cancer transformation, LCNEC-T; large cell neuroendocrine carcinoma transformation. 1 st /2 nd generation EGFR-TKIs include Gefitinib, Erlotinib, Afatinib and Dacomitinib. b Example of longitudinal organoid establishment from a patient with LUAD and SCLC-T (E-02). The computed tomography image at each sampling timepoint is shown on the top, and yellow arrowheads indicate lung cancer lesions. Representative images of hematoxylin and eosin (H&E) staining of the organoids and xenografted tumors (bottom). Scale bar: 100 μm. c Representative H&E staining and NKX2-1 and ΔNp63 immunostaining of the primary tumors, organoids and xenografted tumors of the E-12 line with SQ-T. Scale bar: 100 μm. d Representative H&E staining and NKX2-1 and SYP immunostaining of the primary tumors, organoids and xenografted tumors of the E-15 line with LCNEC-T. Scale bar: 100 μm.

    Journal: Nature Communications

    Article Title: Basal-shift transformation leads to EGFR therapy-resistance in human lung adenocarcinoma

    doi: 10.1038/s41467-025-59623-3

    Figure Lengend Snippet: a Summary of ELCOL. Treatment course for patients with clinical resistance to EGFR-TKI is shown on the right. Circle indicates the timing of tissue sampling and organoid derivation and is colored according to the histology. Thick line shows the treatment period. LUAD; lung adenocarcinoma, SQ-T; squamous transformation, SCLC-T; small cell lung cancer transformation, LCNEC-T; large cell neuroendocrine carcinoma transformation. 1 st /2 nd generation EGFR-TKIs include Gefitinib, Erlotinib, Afatinib and Dacomitinib. b Example of longitudinal organoid establishment from a patient with LUAD and SCLC-T (E-02). The computed tomography image at each sampling timepoint is shown on the top, and yellow arrowheads indicate lung cancer lesions. Representative images of hematoxylin and eosin (H&E) staining of the organoids and xenografted tumors (bottom). Scale bar: 100 μm. c Representative H&E staining and NKX2-1 and ΔNp63 immunostaining of the primary tumors, organoids and xenografted tumors of the E-12 line with SQ-T. Scale bar: 100 μm. d Representative H&E staining and NKX2-1 and SYP immunostaining of the primary tumors, organoids and xenografted tumors of the E-15 line with LCNEC-T. Scale bar: 100 μm.

    Article Snippet: For protein detection, antibodies included rabbit anti-TTF1 (abcam, ab76013, 1:50), rabbit anti-β-actin (CST, 4970, 1:50), rabbit anti-phospho EGFR (Thermo, 44788G-3,1:50), rabbit anti-total EGFR (CST, 4267,1:50), rabbit anti-phospho ERK (CST, 9101,1:50) and rabbit anti-total ERK (CST, 9102,1:50), with subsequent labeling by an HRP-conjugated secondary anti-rabbit or anti-mouse antibody.

    Techniques: Sampling, Transformation Assay, Computed Tomography, Staining, Immunostaining

    a Summary of genetic alterations identified in ELCOL ( n = 39 organoid lines). Genes known to be associated with sensitivity to EGFR-TKI are selected. EGFR T790M and C797S mutations that confer resistance to 1 st /2 nd generation EGFR-TKI and Osimertinib, respectively, are shown independently. b Sensitivity of lung cancer organoids to Osimertinib. The organoids were grouped according to the response of the original tumor to Osimertinib. Cell viability is shown as the ratio of ATP abundance between treated and untreated samples in quadruplicate. Clinically sensitive or resistant lines derive from a tumor judged to be PR/CR or PD, respectively, in RECIST. The IC 50 value of each organoid line is shown on the right. Each dot shows one line. * p = 0.0123, t-test (two-sided). Data are shown as mean ± S.D. Source data are provided as a Source Data file. c Apoptosis analysis of E-01A (Osimertinib sensitive) and E-14 organoids (Osimertinib resistant) treated with DMSO or Osimertinib (1 μM, 72 hours) using flow cytometry. The number (%) indicates the proportion of Annexin V-positive cells. d Organoid derivation from the E-01 patient before and after the development of Osimertinib resistance. Yellow arrowheads show the sampled tumors. Scale bar: 100 μm. e Osimertinib sensitivity in pre- (E-01A) and post- (E-01B) Osimertinib lines. 0.1 μM **** p = 1.85 × 10 −6 , 1 μM **** p = 3.12 × 10 −5 , t-test (two-sided). Data are shown as mean ±S.D. The experiment was performed with four technical replicates. f The proportion of organoid lines with genetically defined mechanisms of Osimertinib resistance and histological transformation in Osimertinib-treated lines ( n = 19). Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: Basal-shift transformation leads to EGFR therapy-resistance in human lung adenocarcinoma

    doi: 10.1038/s41467-025-59623-3

    Figure Lengend Snippet: a Summary of genetic alterations identified in ELCOL ( n = 39 organoid lines). Genes known to be associated with sensitivity to EGFR-TKI are selected. EGFR T790M and C797S mutations that confer resistance to 1 st /2 nd generation EGFR-TKI and Osimertinib, respectively, are shown independently. b Sensitivity of lung cancer organoids to Osimertinib. The organoids were grouped according to the response of the original tumor to Osimertinib. Cell viability is shown as the ratio of ATP abundance between treated and untreated samples in quadruplicate. Clinically sensitive or resistant lines derive from a tumor judged to be PR/CR or PD, respectively, in RECIST. The IC 50 value of each organoid line is shown on the right. Each dot shows one line. * p = 0.0123, t-test (two-sided). Data are shown as mean ± S.D. Source data are provided as a Source Data file. c Apoptosis analysis of E-01A (Osimertinib sensitive) and E-14 organoids (Osimertinib resistant) treated with DMSO or Osimertinib (1 μM, 72 hours) using flow cytometry. The number (%) indicates the proportion of Annexin V-positive cells. d Organoid derivation from the E-01 patient before and after the development of Osimertinib resistance. Yellow arrowheads show the sampled tumors. Scale bar: 100 μm. e Osimertinib sensitivity in pre- (E-01A) and post- (E-01B) Osimertinib lines. 0.1 μM **** p = 1.85 × 10 −6 , 1 μM **** p = 3.12 × 10 −5 , t-test (two-sided). Data are shown as mean ±S.D. The experiment was performed with four technical replicates. f The proportion of organoid lines with genetically defined mechanisms of Osimertinib resistance and histological transformation in Osimertinib-treated lines ( n = 19). Source data are provided as a Source Data file.

    Article Snippet: For protein detection, antibodies included rabbit anti-TTF1 (abcam, ab76013, 1:50), rabbit anti-β-actin (CST, 4970, 1:50), rabbit anti-phospho EGFR (Thermo, 44788G-3,1:50), rabbit anti-total EGFR (CST, 4267,1:50), rabbit anti-phospho ERK (CST, 9101,1:50) and rabbit anti-total ERK (CST, 9102,1:50), with subsequent labeling by an HRP-conjugated secondary anti-rabbit or anti-mouse antibody.

    Techniques: Flow Cytometry, Transformation Assay

    a Principal component analysis of EGFR -mutant lung cancer organoid transcriptomes. The transcriptomes of primary SQ ( n = 5) and SCLC lines ( n = 5) were included for reference. b Genes differentially expressed between primary SQ ( n = 5), SQ-T ( n = 1), pre-EGFR-TKIs LUAD ( n = 7) and post-EGFR-TKIs LUAD without genetic lesions ( n = 11) (FDR < 5.0 ×10 −5 in DESeq2). Expressions in post-EGFR-TKIs LUAD lines without known genetic lesions are also shown. SQ and LUAD genes are co-expressed in LUAD without genetic lesions. c SQ-related gene scores for EGFR -mutant organoid lines. Pre-EGFR-TKIs ( n = 7 lines), LUAD with genetic lesions ( n = 12 lines), LUAD without genetic lesions ( n = 11 lines), SQ-T ( n = 1 line), SQ ( n = 5 lines). Each dot shows SQ-related gene scores for one organoid line. Box plots represent the median (center line), upper and lower quartiles (box limits) and 1.5× interquartile range (whiskers). Source data are provided as a Source Data file. d Coverage of SQ specific ATAC peaks in EGFR -mutant organoid lines. The mean coverage of each peak is shown per subtype. SQ_up indicates upregulated genes in SQ (red). SQ_down indicates downregulated genes in SQ (blue). e scRNA-seq analysis and UMAP embedding of treatment-naïve LUAD, intermediate LUAD and SQ(-T) organoids. The samples were integrated using Harmony. The cell numbers for the UMAP plot are 1797 for E-01A (pre EGFR-TKIs), 4574 for E-05 (pre EGFR-TKIs), 3740 for E-14 (intermediate LUAD), 6924 for E-17 (intermediate LUAD), 1338 for E-20 (intermediate LUAD), 3280 for KOR386 (SQ), 4710 for KOR484 (SQ) and 6777 for E-12 (SQ-T). f Immunofluorescence TAp63 and AQP5 staining in E-13, E-14, E-17, E-20 and E-23 organoid lines with EGFR-TKI resistance and without known genetic lesions. Scale bar: 50 μm. g Representative H&E staining and NKX2-1, ΔNp63 and AQP5 immunostaining of E-23 patient tumors. Scale bar: 100 μm.

    Journal: Nature Communications

    Article Title: Basal-shift transformation leads to EGFR therapy-resistance in human lung adenocarcinoma

    doi: 10.1038/s41467-025-59623-3

    Figure Lengend Snippet: a Principal component analysis of EGFR -mutant lung cancer organoid transcriptomes. The transcriptomes of primary SQ ( n = 5) and SCLC lines ( n = 5) were included for reference. b Genes differentially expressed between primary SQ ( n = 5), SQ-T ( n = 1), pre-EGFR-TKIs LUAD ( n = 7) and post-EGFR-TKIs LUAD without genetic lesions ( n = 11) (FDR < 5.0 ×10 −5 in DESeq2). Expressions in post-EGFR-TKIs LUAD lines without known genetic lesions are also shown. SQ and LUAD genes are co-expressed in LUAD without genetic lesions. c SQ-related gene scores for EGFR -mutant organoid lines. Pre-EGFR-TKIs ( n = 7 lines), LUAD with genetic lesions ( n = 12 lines), LUAD without genetic lesions ( n = 11 lines), SQ-T ( n = 1 line), SQ ( n = 5 lines). Each dot shows SQ-related gene scores for one organoid line. Box plots represent the median (center line), upper and lower quartiles (box limits) and 1.5× interquartile range (whiskers). Source data are provided as a Source Data file. d Coverage of SQ specific ATAC peaks in EGFR -mutant organoid lines. The mean coverage of each peak is shown per subtype. SQ_up indicates upregulated genes in SQ (red). SQ_down indicates downregulated genes in SQ (blue). e scRNA-seq analysis and UMAP embedding of treatment-naïve LUAD, intermediate LUAD and SQ(-T) organoids. The samples were integrated using Harmony. The cell numbers for the UMAP plot are 1797 for E-01A (pre EGFR-TKIs), 4574 for E-05 (pre EGFR-TKIs), 3740 for E-14 (intermediate LUAD), 6924 for E-17 (intermediate LUAD), 1338 for E-20 (intermediate LUAD), 3280 for KOR386 (SQ), 4710 for KOR484 (SQ) and 6777 for E-12 (SQ-T). f Immunofluorescence TAp63 and AQP5 staining in E-13, E-14, E-17, E-20 and E-23 organoid lines with EGFR-TKI resistance and without known genetic lesions. Scale bar: 50 μm. g Representative H&E staining and NKX2-1, ΔNp63 and AQP5 immunostaining of E-23 patient tumors. Scale bar: 100 μm.

    Article Snippet: For protein detection, antibodies included rabbit anti-TTF1 (abcam, ab76013, 1:50), rabbit anti-β-actin (CST, 4970, 1:50), rabbit anti-phospho EGFR (Thermo, 44788G-3,1:50), rabbit anti-total EGFR (CST, 4267,1:50), rabbit anti-phospho ERK (CST, 9101,1:50) and rabbit anti-total ERK (CST, 9102,1:50), with subsequent labeling by an HRP-conjugated secondary anti-rabbit or anti-mouse antibody.

    Techniques: Mutagenesis, Immunofluorescence, Staining, Immunostaining

    a Bar graphs showing the proportion of CDKN2A/B loss in basal-shift ( n = 5) and non-basal-shift ( n = 17) EGFR -mutant organoid lines. Source data are provided as a Source Data file. b Representative pictures of immunohistochemistry for CDKN2A protein in the E-12 and E-23 primary tissues before EGFR-TKI treatment. Scale bar: 100 μm. c Enriched and de-enriched motifs in the basal-shift LUAD lines ( n = 5) compared to the pre-treatment LUAD lines ( n = 6). The motifs were ranked based on the adjusted p values in motif enrichment analysis. d NKX2-1 staining in E-13, E-14, E-17, E-20 and E-23 organoid lines. LUAD and SQ-T lines were used as positive and negative controls, respectively. Scale bar: 100 μm. e Additional knockout of NKX2-1(N) in TP53 (T) and CDKN2A (C) double knockout human alveolar organoids. Confirmation of NKX2-1 knockout in TCN organoids by sanger sequencing (bottom). f Representative NKX2-1 and TAp63 staining in normal alveolar, alveolar_TC and alveolar_TCN lines. Scale bar: 100 μm. At least five independent organoids were evaluated with similar results. g Immunofluorescence staining of TAp63 and AQP5 in an alveolar_TCN organoid. Scale bar: 50 μm. At least five independent organoids were evaluated with similar results. h PCA analysis of lung cancer organoid transcriptomes including genetically engineered alveolar organoids (alveolar_TC and alveolar_TCN). i The growth of the indicated organoid lines in the complete medium containing EGF (E), IGF-1 (I) and FGF-2 (F), and the condition without EIF. The figure indicates the organoid area relative to the control (complete). The experiments were performed in quadruplicate for each condition, technical replicates. Scale bar: 1 mm. Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: Basal-shift transformation leads to EGFR therapy-resistance in human lung adenocarcinoma

    doi: 10.1038/s41467-025-59623-3

    Figure Lengend Snippet: a Bar graphs showing the proportion of CDKN2A/B loss in basal-shift ( n = 5) and non-basal-shift ( n = 17) EGFR -mutant organoid lines. Source data are provided as a Source Data file. b Representative pictures of immunohistochemistry for CDKN2A protein in the E-12 and E-23 primary tissues before EGFR-TKI treatment. Scale bar: 100 μm. c Enriched and de-enriched motifs in the basal-shift LUAD lines ( n = 5) compared to the pre-treatment LUAD lines ( n = 6). The motifs were ranked based on the adjusted p values in motif enrichment analysis. d NKX2-1 staining in E-13, E-14, E-17, E-20 and E-23 organoid lines. LUAD and SQ-T lines were used as positive and negative controls, respectively. Scale bar: 100 μm. e Additional knockout of NKX2-1(N) in TP53 (T) and CDKN2A (C) double knockout human alveolar organoids. Confirmation of NKX2-1 knockout in TCN organoids by sanger sequencing (bottom). f Representative NKX2-1 and TAp63 staining in normal alveolar, alveolar_TC and alveolar_TCN lines. Scale bar: 100 μm. At least five independent organoids were evaluated with similar results. g Immunofluorescence staining of TAp63 and AQP5 in an alveolar_TCN organoid. Scale bar: 50 μm. At least five independent organoids were evaluated with similar results. h PCA analysis of lung cancer organoid transcriptomes including genetically engineered alveolar organoids (alveolar_TC and alveolar_TCN). i The growth of the indicated organoid lines in the complete medium containing EGF (E), IGF-1 (I) and FGF-2 (F), and the condition without EIF. The figure indicates the organoid area relative to the control (complete). The experiments were performed in quadruplicate for each condition, technical replicates. Scale bar: 1 mm. Source data are provided as a Source Data file.

    Article Snippet: For protein detection, antibodies included rabbit anti-TTF1 (abcam, ab76013, 1:50), rabbit anti-β-actin (CST, 4970, 1:50), rabbit anti-phospho EGFR (Thermo, 44788G-3,1:50), rabbit anti-total EGFR (CST, 4267,1:50), rabbit anti-phospho ERK (CST, 9101,1:50) and rabbit anti-total ERK (CST, 9102,1:50), with subsequent labeling by an HRP-conjugated secondary anti-rabbit or anti-mouse antibody.

    Techniques: Mutagenesis, Immunohistochemistry, Staining, Knock-Out, Double Knockout, Sequencing, Immunofluorescence, Control

    a A heatmap showing the therapeutic landscape of 54 compounds in basal-shift (E-14, E-17 and E-23) and non-basal-shift organoid (E-01B and E-16) lines. Percent growth inhibition for each compound versus DMSO is shown in triplicate. Source data are provided as a Source Data file. b IC 50 values of Palbociclib for the four distinct groups: (1) non-basal-shift organoids with CDKN2A/B WT, (2) basal-shift organoids with CDKN2A/B WT, (3) non-basal-shift organoids with CDKN2A/B loss, (4) basal-shift organoids with CDKN2A/B loss. Each dot shows one line. Data are shown as mean ± S.D. Each dot represents the IC 50 value for each line obtained from four technical replicates. Source data are provided as a Source Data file. c Overview of Palbociclib treatment in mouse xenografts. d The effect of Palbociclib on xenografts of CDKN2A/B loss (E-12, E-14 and E-17) or CDKN2A/B WT (E-01B) organoids. n = 5 tumors for each condition. Data are shown as mean ± SD. E-12 ** p = 0.0021, E-14 *** p = 0.0008, E-17 *** p = 0.0004, t-test (two-sided). N.S. not significant. Source data are provided as a Source Data file. e Mechanisms of Osimertinib resistance in ELCOL incorporating basal-shift. Source data are provided as a Source Data file. f The role of basal-shift and other mechanisms in the acquisition of EGFR-TKI resistance.

    Journal: Nature Communications

    Article Title: Basal-shift transformation leads to EGFR therapy-resistance in human lung adenocarcinoma

    doi: 10.1038/s41467-025-59623-3

    Figure Lengend Snippet: a A heatmap showing the therapeutic landscape of 54 compounds in basal-shift (E-14, E-17 and E-23) and non-basal-shift organoid (E-01B and E-16) lines. Percent growth inhibition for each compound versus DMSO is shown in triplicate. Source data are provided as a Source Data file. b IC 50 values of Palbociclib for the four distinct groups: (1) non-basal-shift organoids with CDKN2A/B WT, (2) basal-shift organoids with CDKN2A/B WT, (3) non-basal-shift organoids with CDKN2A/B loss, (4) basal-shift organoids with CDKN2A/B loss. Each dot shows one line. Data are shown as mean ± S.D. Each dot represents the IC 50 value for each line obtained from four technical replicates. Source data are provided as a Source Data file. c Overview of Palbociclib treatment in mouse xenografts. d The effect of Palbociclib on xenografts of CDKN2A/B loss (E-12, E-14 and E-17) or CDKN2A/B WT (E-01B) organoids. n = 5 tumors for each condition. Data are shown as mean ± SD. E-12 ** p = 0.0021, E-14 *** p = 0.0008, E-17 *** p = 0.0004, t-test (two-sided). N.S. not significant. Source data are provided as a Source Data file. e Mechanisms of Osimertinib resistance in ELCOL incorporating basal-shift. Source data are provided as a Source Data file. f The role of basal-shift and other mechanisms in the acquisition of EGFR-TKI resistance.

    Article Snippet: For protein detection, antibodies included rabbit anti-TTF1 (abcam, ab76013, 1:50), rabbit anti-β-actin (CST, 4970, 1:50), rabbit anti-phospho EGFR (Thermo, 44788G-3,1:50), rabbit anti-total EGFR (CST, 4267,1:50), rabbit anti-phospho ERK (CST, 9101,1:50) and rabbit anti-total ERK (CST, 9102,1:50), with subsequent labeling by an HRP-conjugated secondary anti-rabbit or anti-mouse antibody.

    Techniques: Inhibition

    EGF promotes OSCC progression by activating the EGFR/NF-κB pathway. A Cal27 cells were treated with EGF (100 ng/ml) for 0, 30, 60, and 120 min, and the expression of EGFR was detected by Western blot analysis. B , C Quantified detection of the EGFR expression. D Western blot analysis of the expression of EGFR and members of the NF-κB signaling pathway in Cal27 cells treated with EGF or EGF and AG1478. E , F Quantitative analysis of Western blots in ( D ) Cal27 cells. G mRNA expression of CXCL1 in Cal27 cells treated with EGF (100 ng/ml) or EGF and AG1478 (5 μM) in TAM-CM. H Measurement of CXCL1 levels in treated Cal27 cells by ELISA. I Western blot analysis of the expression of CXCL1 in Cal27 cells treated with EGF or EGF and AG1478-containing TAM-CM. Bar graphs are presented as the mean ± SD (n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001 vs. the control

    Journal: Discover Oncology

    Article Title: Crosstalk between cancer cells and macrophages promotes OSCC cell migration and invasion through a CXCL1/EGF positive feedback loop

    doi: 10.1007/s12672-024-00972-8

    Figure Lengend Snippet: EGF promotes OSCC progression by activating the EGFR/NF-κB pathway. A Cal27 cells were treated with EGF (100 ng/ml) for 0, 30, 60, and 120 min, and the expression of EGFR was detected by Western blot analysis. B , C Quantified detection of the EGFR expression. D Western blot analysis of the expression of EGFR and members of the NF-κB signaling pathway in Cal27 cells treated with EGF or EGF and AG1478. E , F Quantitative analysis of Western blots in ( D ) Cal27 cells. G mRNA expression of CXCL1 in Cal27 cells treated with EGF (100 ng/ml) or EGF and AG1478 (5 μM) in TAM-CM. H Measurement of CXCL1 levels in treated Cal27 cells by ELISA. I Western blot analysis of the expression of CXCL1 in Cal27 cells treated with EGF or EGF and AG1478-containing TAM-CM. Bar graphs are presented as the mean ± SD (n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001 vs. the control

    Article Snippet: The antibodies used included rabbit anti-β-actin (cat. no. ab115777; Abcam), rabbit anti-E-cadherin (cat. no. ab40772 Abcam), rabbit anti-N-cadherin (cat. no. ab76011 Abcam), rabbit anti-CXCL1 (cat. no. ab206411 Abcam), rabbit anti-total EGFR (Zen Bio Science Co., Ltd), rabbit anti-phospho-EGFR (cat. no. R24173; Zen Bio Science Co., Ltd), rabbit anti-total P65 (cat. no. R25149; Zen Bio Science Co., Ltd), and rabbit anti-phospho-P65 (cat. no. 310013; Zen Bio Science Co., Ltd).

    Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay

    Model of crosstalk between cancer cells and activated macrophages promoting OSCC progression via the CXCL1/EGF feedback loop. Tumor-derived CXCL1 facilitates the secretion of EGF in TAMs, while TAM-derived EGF induces the activation of EGFR/NF-κB in OSCC cells, which accelerates tumor cell migration and invasion

    Journal: Discover Oncology

    Article Title: Crosstalk between cancer cells and macrophages promotes OSCC cell migration and invasion through a CXCL1/EGF positive feedback loop

    doi: 10.1007/s12672-024-00972-8

    Figure Lengend Snippet: Model of crosstalk between cancer cells and activated macrophages promoting OSCC progression via the CXCL1/EGF feedback loop. Tumor-derived CXCL1 facilitates the secretion of EGF in TAMs, while TAM-derived EGF induces the activation of EGFR/NF-κB in OSCC cells, which accelerates tumor cell migration and invasion

    Article Snippet: The antibodies used included rabbit anti-β-actin (cat. no. ab115777; Abcam), rabbit anti-E-cadherin (cat. no. ab40772 Abcam), rabbit anti-N-cadherin (cat. no. ab76011 Abcam), rabbit anti-CXCL1 (cat. no. ab206411 Abcam), rabbit anti-total EGFR (Zen Bio Science Co., Ltd), rabbit anti-phospho-EGFR (cat. no. R24173; Zen Bio Science Co., Ltd), rabbit anti-total P65 (cat. no. R25149; Zen Bio Science Co., Ltd), and rabbit anti-phospho-P65 (cat. no. 310013; Zen Bio Science Co., Ltd).

    Techniques: Derivative Assay, Activation Assay, Migration