bon 1 cells  (ATCC)

Journal: Cell Reports Medicine

Article Title: Targeting the ferritinophagy-lysosome axis as a therapeutic vulnerability in gastroenteropancreatic neuroendocrine tumors

doi: 10.1016/j.xcrm.2026.102695

Figure Lengend Snippet: Kinome-wide CRISPR knockout screen identifies PIKfyve as a druggable target in GEP-NETs (A) Schematic of the CRISPR screening workflow in BON-1-DsRed-IRES-GFP-p62 cells. (B) Gene enrichment rank plot from kinome CRISPR knockout screens (specified in ). Cutoff was set at fold change < −0.3. Red genes are related to the phosphatidylinositol metabolic pathway, while blue genes are related to the mTOR pathway. (C) Pathway enrichment analysis of essential genes from CRISPR screening data in (B) using PANTHER 19.0. Targets with a fold change < −0.3 were considered candidate genes, and pathways with a p value <0.05 and hit targets ≥3 were displayed in the plot. Pathways labeled in red are related to phosphatidylinositol metabolism. (D) Schematic of PI(3,5)P2 biosynthesis regulated by VPS34 ( PIK3C3 ) and PIKfyve. (E) Cell confluence of BON-1 and QGP-1 cells after siRNA-mediated knockdown of PIK3C3 , PIKFYVE , or non-targeting control (siNC). Data are presented as mean ± SD ( n = 4 biological replicates). Two-way ANOVA. (F) Crystal violet staining showing long-term growth inhibition by PIKfyve inhibitor apilimod and mTOR inhibitor (Torin-1 or everolimus) in indicated GEP-NETs cell lines and HPNE cells. (G) Compilation of IC 50 values for everolimus, apilimod, and ESK981 in GEP-NET and HPNE cells. (H and I) Dose-response proliferation curves of everolimus (H) and apilimod (I) in indicated GEP-NET cell lines and HPNE cells. Data are presented as mean ± SD (at least three biological replicates).

Article Snippet: BON-1 cells were cultured in DMEM/F12; STC-1, HT-1080, and Hep G2 were cultured in DMEM; 786-O, QGP-1, and GOT-1 were cultured in ATCC-formulated RPMI-1640; Caki-1 was cultured in McCoy’s 5A.

Techniques: CRISPR, Knock-Out, Labeling, Knockdown, Control, Staining, Inhibition

Journal: Cell Reports Medicine

Article Title: Targeting the ferritinophagy-lysosome axis as a therapeutic vulnerability in gastroenteropancreatic neuroendocrine tumors

doi: 10.1016/j.xcrm.2026.102695

Figure Lengend Snippet: PIKfyve is overexpressed in GEP-NETs and serves as a therapeutic target (A) Representative PIKfyve IHC staining in human normal colon tissue, colon neuroendocrine tumor, and colon adenocarcinoma samples. (B) Quantification of PIKfyve H-score from tissue microarray of human normal GEP tissue, GEP-NETs, and GEP adenocarcinoma (specified in ). Statistics were performed using one-way ANOVA. (C) Immunoblot analysis of PIKfyve and autophagy markers (p62 and LC3A/B) in GEP-NET cell lines following CRISPRi-mediated PIKFYVE knockdown. Vinculin served as the loading control. (D) Cell confluence of GEP-NET cell lines with CRISPRi-mediated PIKFYVE (sg PIKFYVE or sg Pikfyve ) knockdown or control (sgNC). Data are presented as mean ± SD ( n = 4 biological replicates). Two-way ANOVA. (E) Immunoblot analysis of autophagy markers in QGP-1 and BON-1 cells following PIKfyve inhibitors (apilimod or ESK981) or PIKfyve degrader (PIK5-33d) treatment for 8 or 24 h. GAPDH was used as a loading control. (F) Average tumor volumes of QGP-1 subcutaneous-cell-line-derived (CDX) model for vehicle ( n = 9) or ESK981 (30 mg/kg, n = 10) treatment. Mean ± SEM. Two-way ANOVA. s.c., subcutaneous. (G) Spider plot displaying individual tumor volumes from vehicle or ESK981 treatment groups in QGP-1 subcutaneous CDX study. (H) Individual tumor weights of QGP-1 subcutaneous CDX model tumors at study endpoint. Unpaired two-tailed t test. (I) Percent body weight change of QGP-1 subcutaneous CDX model tumors from vehicle or ESK981 treatment groups. (J) Average tumor volumes of BON-1 subcutaneous CDX model following vehicle ( n = 17) or ESK981 (30 mg/kg, n = 15) treatment. Mean ± SEM. Two-way ANOVA. (K) Spider plot displaying individual tumor volumes from vehicle or ESK981 treatment groups in BON-1 subcutaneous CDX study. (L) Individual tumor weights of BON-1 subcutaneous CDX model tumors at study endpoint. Unpaired two-tailed t test. (M) Percent body weight change of BON-1 subcutaneous CDX model tumors from vehicle or ESK981 treatment groups.

Article Snippet: BON-1 cells were cultured in DMEM/F12; STC-1, HT-1080, and Hep G2 were cultured in DMEM; 786-O, QGP-1, and GOT-1 were cultured in ATCC-formulated RPMI-1640; Caki-1 was cultured in McCoy’s 5A.

Techniques: Immunohistochemistry, Microarray, Western Blot, Knockdown, Control, Derivative Assay, Two Tailed Test

Journal: Cell Reports Medicine

Article Title: Targeting the ferritinophagy-lysosome axis as a therapeutic vulnerability in gastroenteropancreatic neuroendocrine tumors

doi: 10.1016/j.xcrm.2026.102695

Figure Lengend Snippet: PIKfyve mediates lipid homeostasis in GEP-NETs (A) Pathway enrichment analysis of RNA-seq from QGP-1 cells following CRISPRi-mediated PIKFYVE knockdown. (B) GSEA of fatty acid metabolism, cholesterol homeostasis, and mTORC1 signaling after CRISPRi-mediated PIKFYVE knockdown in QGP-1 cells. (C) Volcano plot of differentially expressed genes highlighting fatty acid metabolism (violet), cholesterol homeostasis (rose), or mTORC1 signaling pathways (orange). (D) Pathway enrichment analysis of RNA-seq from QGP-1 cells treated with PIKfyve inhibitor apilimod (1 μM, 8 h). (E) GSEA of fatty acid metabolism, cholesterol homeostasis, and mTORC1 signaling following apilimod treatment. (F) Volcano plots of differentially expressed genes from QGP-1 cells after apilimod treatment, highlighting fatty acid metabolism (violet), cholesterol homeostasis (rose), or mTORC1 signaling pathways (orange). (G) Schematic illustrating SREBP- and mTOR-dependent regulation of fatty acid cholesterol biosynthesis. (H) Immunoblot showing PIKfyve, premature SREBP1 (p), mature SREBP1 (m), FASN, and SCD expression in QGP-1 and BON-1 cells following genetic or pharmacological PIKfyve inhibition (inhibitors: apilimod, ESK981; degrader: PIK5-33d. 8-h treatment). GAPDH was used as a loading control. (I) LAMP1 and filipin (cholesterol probe) staining showing lysosomal cholesterol accumulation after apilimod or ESK981 treatment for 24 h at 1 μM. Scale bars: 5 μm. (J and K) Synergy analyses of apilimod and the SCD inhibitor (CAY10566) in QGP-1 (J) and STC-1 (K) cells, shown as dose-response heatmaps and 3D synergy plots.

Article Snippet: BON-1 cells were cultured in DMEM/F12; STC-1, HT-1080, and Hep G2 were cultured in DMEM; 786-O, QGP-1, and GOT-1 were cultured in ATCC-formulated RPMI-1640; Caki-1 was cultured in McCoy’s 5A.

Techniques: RNA Sequencing, Knockdown, Protein-Protein interactions, Western Blot, Expressing, Inhibition, Control, Staining

Journal: Cell Reports Medicine

Article Title: Targeting the ferritinophagy-lysosome axis as a therapeutic vulnerability in gastroenteropancreatic neuroendocrine tumors

doi: 10.1016/j.xcrm.2026.102695

Figure Lengend Snippet: Inhibition of the mTOR pathway suppresses the SREBP1 pathway and triggers ferritinophagy (A) Pathway enrichment analysis of RNA-seq in QGP-1 cells treated with Torin-1 (0.1 μM) for 8 h (B) Volcano plot of differentially expressed proteins from whole-cell proteomics of QGP-1 cells after Torin-1 treatment (0.1 μM, 24 h) (specified in ), highlighting fatty acid metabolism, cholesterol homeostasis, and mTORC1 signaling pathways. (C) Immunoblot analysis of mTOR pathway and lipid metabolism proteins in BON-1 and QGP-1 cells after 24 h of the indicated treatment. GAPDH was used as a loading control. (D) Immunoblot showing autophagy-related proteins in BON-1 and QGP-1 cells following Torin-1 or everolimus treatment for 24 h. GAPDH was used as a loading control. (E) Tandem fluorescent reporter assay assessing autophagic flux in QGP-1 and BON-1 cells treated with Torin-1 (0.1 μM) or everolimus (5 μM) for 24 h. One-way ANOVA. (F) Schematic of TMEM192-labeled cells for lysosome isolation and proteomic analysis. (G) Volcano plot of differentially expressed lysosomal proteins of QGP-1 TMEM192 cells treated with Torin-1 (0.1 μM, 24 h) (specified in ), highlighting proteins in mTORC1 signaling (orange) and ferritinophagy (green). (H) Immunoblot validating increased ferritin levels (FTL and FTH1) in QGP-1 TMEM192 cells following Torin-1 treatment for 24 h. GAPDH was used as a loading control for whole-cell lysates, while LAMP1 served as a loading control for lysosomal samples. (I and J) Immunoblot analysis of ferritin and transferrin levels in BON-1 and QGP-1 cells treated with mTOR inhibitor (Torin-1, everolimus) for 24 h or siRNA-mediated knockdown of mTOR. GAPDH and β-actin were used as loading controls. (K) Representative image of FerroOrange staining in QGP-1 cells with the indicated compounds. Torin-1 was used at 100 nM, and Ferrous ammonium sulfate (FAS) was used as a positive control (50 μM). Scale bars: 20 μm. (L) Intracellular iron levels measured using FerroOrange dye via flow cytometry in QGP-1 cells treated with the indicated compounds. Torin-1 was used at 100 nM, and FAS was used at 100 μM. One-way ANOVA. (M) Confluence assay showing enhanced growth inhibition by everolimus (5 μM) combined with iron chelating agent deferoxamine (DFO, 20 μM). Data are presented as mean ± SD ( n = 3∼4). Two-way ANOVA.

Article Snippet: BON-1 cells were cultured in DMEM/F12; STC-1, HT-1080, and Hep G2 were cultured in DMEM; 786-O, QGP-1, and GOT-1 were cultured in ATCC-formulated RPMI-1640; Caki-1 was cultured in McCoy’s 5A.

Techniques: Inhibition, RNA Sequencing, Protein-Protein interactions, Western Blot, Control, Reporter Assay, Labeling, Isolation, Knockdown, Staining, Positive Control, Flow Cytometry

Journal: Cell Reports Medicine

Article Title: Targeting the ferritinophagy-lysosome axis as a therapeutic vulnerability in gastroenteropancreatic neuroendocrine tumors

doi: 10.1016/j.xcrm.2026.102695

Figure Lengend Snippet: PIKfyve blockade abrogates mTOR-inhibition-induced ferritinophagy (A and B) Autophagic flux in QGP-1 (A) and BON-1 (B) cells treated with DMSO, apilimod (AP), ESK981 (ESK), or combinations with Torin-1 (0.1 μM) or everolimus (5 μM) for 24 h. Statistical analysis using one-way ANOVA. (C) Immunoblot analysis of mTOR signaling and autophagy markers in QGP-1 and BON-1 cells treated with mTOR inhibitors with or without PIKfyve antagonists. GAPDH served as a loading control. (D) Volcano plot of differentially expressed lysosomal proteins in QGP-1 TMEM192 cells treated with or without apilimod (1 μM, 24 h) (specified in ). (E) Pathway enrichment analysis of lysosomal proteomics highlighting ferritinophagy-related pathways. (F) Immunoblot validation of decreased ferritin levels (FTL and FTH1) in QGP-1 TMEM192 cells following the indicated treatment for 24 h. GAPDH served as loading control for whole-cell lysates; LAMP1 served as loading control for lysosomal samples. (G and H) Intracellular iron levels in QGP-1 cells assessed by FerroOrange staining (G) and flow cytometry (H). FAS served as a positive control; deferoxamine (DFO) served as a negative control. Scale bars: 20 μm. Statistical analysis using two-way ANOVA. (I) Immunoblot analysis of SDHB following changes in bioavailable iron after lysosomal or PIKfyve inhibition with or without ammonium ferric citrate (FAC) supplementation in indicated cells. β-actin served as the loading control. (J) Confluence assay showing FAC (100 μg/mL)-mediated rescue of cell growth after apilimod treatment. All conditions were supplemented with 1 μM Ferrostatin-1 to prevent the deleterious effects of excess free iron. Data presented as mean ± SD ( n = 3). Statistical analysis using two-way ANOVA.

Article Snippet: BON-1 cells were cultured in DMEM/F12; STC-1, HT-1080, and Hep G2 were cultured in DMEM; 786-O, QGP-1, and GOT-1 were cultured in ATCC-formulated RPMI-1640; Caki-1 was cultured in McCoy’s 5A.

Techniques: Inhibition, Western Blot, Control, Biomarker Discovery, Staining, Flow Cytometry, Positive Control, Negative Control

Journal: Cell Reports Medicine

Article Title: Targeting the ferritinophagy-lysosome axis as a therapeutic vulnerability in gastroenteropancreatic neuroendocrine tumors

doi: 10.1016/j.xcrm.2026.102695

Figure Lengend Snippet: Dual inhibition of mTOR and PIKfyve triggers synthetic lethality in vitro (A) Heatmap showing RT-qPCR analysis of lipid metabolism targets in BON-1 cells treated with mTOR inhibitors (Torin: Torin-1; Evero: everolimus) with or without PIKfyve antagonists. Statistical analysis using two-way ANOVA. (B) Immunoblot analysis of lipid metabolism proteins in QGP-1 cells treated as (A). GAPDH, loading control. (C) Everolimus IC 50 curves in QGP-1 cells with or without CRISPRi-mediated PIKFYVE knockdown. Inset shows IC 50 values. (D) Confluence assay showing the efficacy of everolimus (5 μM) upon PIKFYVE knockdown in QGP-1 cells. Data shown are mean ± SD ( n = 3). Two-way ANOVA. (E and F) 3D synergy plots and heatmaps for QGP-1 cells treated with everolimus and apilimod or ESK981. Red peaks indicate synergy, with the average synergy score shown. (G and H) Confluence assay showing synergistic effect of apilimod (1 μM) or ESK981 (250 nM) combined with everolimus (5 μM). Data presented as mean ± SD ( n = 3). Statistical analysis using two-way ANOVA. (I) 3D synergy plots and heatmaps for apilimod and everolimus with rescue by DMSO, Ferrostatin-1 (1 μM), or Z-VAD-FMK (10 μM). (J) Confluence assay showing synergistic effect of apilimod (1 μM) and everolimus (5 μM) rescued with DMSO, Ferrostatin-1 (1 μM), or Z-VAD-FMK (10 μM). Data presented as mean ± SD ( n = 3). Statistical analysis using two-way ANOVA.

Article Snippet: BON-1 cells were cultured in DMEM/F12; STC-1, HT-1080, and Hep G2 were cultured in DMEM; 786-O, QGP-1, and GOT-1 were cultured in ATCC-formulated RPMI-1640; Caki-1 was cultured in McCoy’s 5A.

Techniques: Inhibition, In Vitro, Quantitative RT-PCR, Western Blot, Control, Knockdown

Journal: Cell Reports Medicine

Article Title: Targeting the ferritinophagy-lysosome axis as a therapeutic vulnerability in gastroenteropancreatic neuroendocrine tumors

doi: 10.1016/j.xcrm.2026.102695

Figure Lengend Snippet: Combinatorial targeting of mTOR and PIKfyve exerts synergistic effects in vivo in GEP-NETs (A) Immunoblot of QGP-1 and BON-1 CDX tumors after 5 days (PD5) of treatment with vehicle, ESK981 (30 mg/kg), everolimus (2.5 mg/kg), or the combination, showing cleaved PARP (c-PARP); GAPDH, loading control. (B) Schematic of orthotopic pancreatic neuroendocrine tumor model (QGP-1 or BON-1) and treatment regimens. (C) Tumor-to-body weight ratio of pancreas from QGP-1 orthotopic model. (D) Percentage normal pancreas area from the study in (C). Data presented as mean ± SEM. p values calculated using one-way ANOVA. (E) Bioluminescence imaging (BLI) of QGP-1 orthotopic tumors across treatment groups. (F) Kaplan-Meier survival curves of QGP-1 tumor-bearing mice. Data expressed as means ± SEM. p values calculated using two-way repeated-measures ANOVA. (G) Percentage body weight changes of QGP-1 tumor-bearing mice. Data presented as mean ± SEM. (H) BLI of BON-1 orthotopic tumors. Data presented as mean ± SEM. (I) Percentage change in BLI signal on day 28 compared to day 0 for BON-1 orthotopic model shown in (H). (J) Kaplan-Meier survival curves of BON-1 tumor-bearing mice. Data expressed as means ± SEM. p values calculated using two-way repeated-measures ANOVA.

Article Snippet: BON-1 cells were cultured in DMEM/F12; STC-1, HT-1080, and Hep G2 were cultured in DMEM; 786-O, QGP-1, and GOT-1 were cultured in ATCC-formulated RPMI-1640; Caki-1 was cultured in McCoy’s 5A.

Techniques: In Vivo, Western Blot, Control, Imaging

Journal: Cell Reports Medicine

Article Title: Targeting the ferritinophagy-lysosome axis as a therapeutic vulnerability in gastroenteropancreatic neuroendocrine tumors

doi: 10.1016/j.xcrm.2026.102695

Figure Lengend Snippet: Kinome-wide CRISPR knockout screen identifies PIKfyve as a druggable target in GEP-NETs (A) Schematic of the CRISPR screening workflow in BON-1-DsRed-IRES-GFP-p62 cells. (B) Gene enrichment rank plot from kinome CRISPR knockout screens (specified in ). Cutoff was set at fold change < −0.3. Red genes are related to the phosphatidylinositol metabolic pathway, while blue genes are related to the mTOR pathway. (C) Pathway enrichment analysis of essential genes from CRISPR screening data in (B) using PANTHER 19.0. Targets with a fold change < −0.3 were considered candidate genes, and pathways with a p value <0.05 and hit targets ≥3 were displayed in the plot. Pathways labeled in red are related to phosphatidylinositol metabolism. (D) Schematic of PI(3,5)P2 biosynthesis regulated by VPS34 ( PIK3C3 ) and PIKfyve. (E) Cell confluence of BON-1 and QGP-1 cells after siRNA-mediated knockdown of PIK3C3 , PIKFYVE , or non-targeting control (siNC). Data are presented as mean ± SD ( n = 4 biological replicates). Two-way ANOVA. (F) Crystal violet staining showing long-term growth inhibition by PIKfyve inhibitor apilimod and mTOR inhibitor (Torin-1 or everolimus) in indicated GEP-NETs cell lines and HPNE cells. (G) Compilation of IC 50 values for everolimus, apilimod, and ESK981 in GEP-NET and HPNE cells. (H and I) Dose-response proliferation curves of everolimus (H) and apilimod (I) in indicated GEP-NET cell lines and HPNE cells. Data are presented as mean ± SD (at least three biological replicates).

Article Snippet: BON-1 was purchased from Creative Biolabs.

Techniques: CRISPR, Knock-Out, Labeling, Knockdown, Control, Staining, Inhibition

Journal: Cell Reports Medicine

Article Title: Targeting the ferritinophagy-lysosome axis as a therapeutic vulnerability in gastroenteropancreatic neuroendocrine tumors

doi: 10.1016/j.xcrm.2026.102695

Figure Lengend Snippet: PIKfyve is overexpressed in GEP-NETs and serves as a therapeutic target (A) Representative PIKfyve IHC staining in human normal colon tissue, colon neuroendocrine tumor, and colon adenocarcinoma samples. (B) Quantification of PIKfyve H-score from tissue microarray of human normal GEP tissue, GEP-NETs, and GEP adenocarcinoma (specified in ). Statistics were performed using one-way ANOVA. (C) Immunoblot analysis of PIKfyve and autophagy markers (p62 and LC3A/B) in GEP-NET cell lines following CRISPRi-mediated PIKFYVE knockdown. Vinculin served as the loading control. (D) Cell confluence of GEP-NET cell lines with CRISPRi-mediated PIKFYVE (sg PIKFYVE or sg Pikfyve ) knockdown or control (sgNC). Data are presented as mean ± SD ( n = 4 biological replicates). Two-way ANOVA. (E) Immunoblot analysis of autophagy markers in QGP-1 and BON-1 cells following PIKfyve inhibitors (apilimod or ESK981) or PIKfyve degrader (PIK5-33d) treatment for 8 or 24 h. GAPDH was used as a loading control. (F) Average tumor volumes of QGP-1 subcutaneous-cell-line-derived (CDX) model for vehicle ( n = 9) or ESK981 (30 mg/kg, n = 10) treatment. Mean ± SEM. Two-way ANOVA. s.c., subcutaneous. (G) Spider plot displaying individual tumor volumes from vehicle or ESK981 treatment groups in QGP-1 subcutaneous CDX study. (H) Individual tumor weights of QGP-1 subcutaneous CDX model tumors at study endpoint. Unpaired two-tailed t test. (I) Percent body weight change of QGP-1 subcutaneous CDX model tumors from vehicle or ESK981 treatment groups. (J) Average tumor volumes of BON-1 subcutaneous CDX model following vehicle ( n = 17) or ESK981 (30 mg/kg, n = 15) treatment. Mean ± SEM. Two-way ANOVA. (K) Spider plot displaying individual tumor volumes from vehicle or ESK981 treatment groups in BON-1 subcutaneous CDX study. (L) Individual tumor weights of BON-1 subcutaneous CDX model tumors at study endpoint. Unpaired two-tailed t test. (M) Percent body weight change of BON-1 subcutaneous CDX model tumors from vehicle or ESK981 treatment groups.

Article Snippet: BON-1 was purchased from Creative Biolabs.

Techniques: Immunohistochemistry, Microarray, Western Blot, Knockdown, Control, Derivative Assay, Two Tailed Test

Journal: Cell Reports Medicine

Article Title: Targeting the ferritinophagy-lysosome axis as a therapeutic vulnerability in gastroenteropancreatic neuroendocrine tumors

doi: 10.1016/j.xcrm.2026.102695

Figure Lengend Snippet: PIKfyve mediates lipid homeostasis in GEP-NETs (A) Pathway enrichment analysis of RNA-seq from QGP-1 cells following CRISPRi-mediated PIKFYVE knockdown. (B) GSEA of fatty acid metabolism, cholesterol homeostasis, and mTORC1 signaling after CRISPRi-mediated PIKFYVE knockdown in QGP-1 cells. (C) Volcano plot of differentially expressed genes highlighting fatty acid metabolism (violet), cholesterol homeostasis (rose), or mTORC1 signaling pathways (orange). (D) Pathway enrichment analysis of RNA-seq from QGP-1 cells treated with PIKfyve inhibitor apilimod (1 μM, 8 h). (E) GSEA of fatty acid metabolism, cholesterol homeostasis, and mTORC1 signaling following apilimod treatment. (F) Volcano plots of differentially expressed genes from QGP-1 cells after apilimod treatment, highlighting fatty acid metabolism (violet), cholesterol homeostasis (rose), or mTORC1 signaling pathways (orange). (G) Schematic illustrating SREBP- and mTOR-dependent regulation of fatty acid cholesterol biosynthesis. (H) Immunoblot showing PIKfyve, premature SREBP1 (p), mature SREBP1 (m), FASN, and SCD expression in QGP-1 and BON-1 cells following genetic or pharmacological PIKfyve inhibition (inhibitors: apilimod, ESK981; degrader: PIK5-33d. 8-h treatment). GAPDH was used as a loading control. (I) LAMP1 and filipin (cholesterol probe) staining showing lysosomal cholesterol accumulation after apilimod or ESK981 treatment for 24 h at 1 μM. Scale bars: 5 μm. (J and K) Synergy analyses of apilimod and the SCD inhibitor (CAY10566) in QGP-1 (J) and STC-1 (K) cells, shown as dose-response heatmaps and 3D synergy plots.

Article Snippet: BON-1 was purchased from Creative Biolabs.

Techniques: RNA Sequencing, Knockdown, Protein-Protein interactions, Western Blot, Expressing, Inhibition, Control, Staining

Journal: Cell Reports Medicine

Article Title: Targeting the ferritinophagy-lysosome axis as a therapeutic vulnerability in gastroenteropancreatic neuroendocrine tumors

doi: 10.1016/j.xcrm.2026.102695

Figure Lengend Snippet: Inhibition of the mTOR pathway suppresses the SREBP1 pathway and triggers ferritinophagy (A) Pathway enrichment analysis of RNA-seq in QGP-1 cells treated with Torin-1 (0.1 μM) for 8 h (B) Volcano plot of differentially expressed proteins from whole-cell proteomics of QGP-1 cells after Torin-1 treatment (0.1 μM, 24 h) (specified in ), highlighting fatty acid metabolism, cholesterol homeostasis, and mTORC1 signaling pathways. (C) Immunoblot analysis of mTOR pathway and lipid metabolism proteins in BON-1 and QGP-1 cells after 24 h of the indicated treatment. GAPDH was used as a loading control. (D) Immunoblot showing autophagy-related proteins in BON-1 and QGP-1 cells following Torin-1 or everolimus treatment for 24 h. GAPDH was used as a loading control. (E) Tandem fluorescent reporter assay assessing autophagic flux in QGP-1 and BON-1 cells treated with Torin-1 (0.1 μM) or everolimus (5 μM) for 24 h. One-way ANOVA. (F) Schematic of TMEM192-labeled cells for lysosome isolation and proteomic analysis. (G) Volcano plot of differentially expressed lysosomal proteins of QGP-1 TMEM192 cells treated with Torin-1 (0.1 μM, 24 h) (specified in ), highlighting proteins in mTORC1 signaling (orange) and ferritinophagy (green). (H) Immunoblot validating increased ferritin levels (FTL and FTH1) in QGP-1 TMEM192 cells following Torin-1 treatment for 24 h. GAPDH was used as a loading control for whole-cell lysates, while LAMP1 served as a loading control for lysosomal samples. (I and J) Immunoblot analysis of ferritin and transferrin levels in BON-1 and QGP-1 cells treated with mTOR inhibitor (Torin-1, everolimus) for 24 h or siRNA-mediated knockdown of mTOR. GAPDH and β-actin were used as loading controls. (K) Representative image of FerroOrange staining in QGP-1 cells with the indicated compounds. Torin-1 was used at 100 nM, and Ferrous ammonium sulfate (FAS) was used as a positive control (50 μM). Scale bars: 20 μm. (L) Intracellular iron levels measured using FerroOrange dye via flow cytometry in QGP-1 cells treated with the indicated compounds. Torin-1 was used at 100 nM, and FAS was used at 100 μM. One-way ANOVA. (M) Confluence assay showing enhanced growth inhibition by everolimus (5 μM) combined with iron chelating agent deferoxamine (DFO, 20 μM). Data are presented as mean ± SD ( n = 3∼4). Two-way ANOVA.

Article Snippet: BON-1 was purchased from Creative Biolabs.

Techniques: Inhibition, RNA Sequencing, Protein-Protein interactions, Western Blot, Control, Reporter Assay, Labeling, Isolation, Knockdown, Staining, Positive Control, Flow Cytometry

Journal: Cell Reports Medicine

Article Title: Targeting the ferritinophagy-lysosome axis as a therapeutic vulnerability in gastroenteropancreatic neuroendocrine tumors

doi: 10.1016/j.xcrm.2026.102695

Figure Lengend Snippet: PIKfyve blockade abrogates mTOR-inhibition-induced ferritinophagy (A and B) Autophagic flux in QGP-1 (A) and BON-1 (B) cells treated with DMSO, apilimod (AP), ESK981 (ESK), or combinations with Torin-1 (0.1 μM) or everolimus (5 μM) for 24 h. Statistical analysis using one-way ANOVA. (C) Immunoblot analysis of mTOR signaling and autophagy markers in QGP-1 and BON-1 cells treated with mTOR inhibitors with or without PIKfyve antagonists. GAPDH served as a loading control. (D) Volcano plot of differentially expressed lysosomal proteins in QGP-1 TMEM192 cells treated with or without apilimod (1 μM, 24 h) (specified in ). (E) Pathway enrichment analysis of lysosomal proteomics highlighting ferritinophagy-related pathways. (F) Immunoblot validation of decreased ferritin levels (FTL and FTH1) in QGP-1 TMEM192 cells following the indicated treatment for 24 h. GAPDH served as loading control for whole-cell lysates; LAMP1 served as loading control for lysosomal samples. (G and H) Intracellular iron levels in QGP-1 cells assessed by FerroOrange staining (G) and flow cytometry (H). FAS served as a positive control; deferoxamine (DFO) served as a negative control. Scale bars: 20 μm. Statistical analysis using two-way ANOVA. (I) Immunoblot analysis of SDHB following changes in bioavailable iron after lysosomal or PIKfyve inhibition with or without ammonium ferric citrate (FAC) supplementation in indicated cells. β-actin served as the loading control. (J) Confluence assay showing FAC (100 μg/mL)-mediated rescue of cell growth after apilimod treatment. All conditions were supplemented with 1 μM Ferrostatin-1 to prevent the deleterious effects of excess free iron. Data presented as mean ± SD ( n = 3). Statistical analysis using two-way ANOVA.

Article Snippet: BON-1 was purchased from Creative Biolabs.

Techniques: Inhibition, Western Blot, Control, Biomarker Discovery, Staining, Flow Cytometry, Positive Control, Negative Control

Journal: Cell Reports Medicine

Article Title: Targeting the ferritinophagy-lysosome axis as a therapeutic vulnerability in gastroenteropancreatic neuroendocrine tumors

doi: 10.1016/j.xcrm.2026.102695

Figure Lengend Snippet: Dual inhibition of mTOR and PIKfyve triggers synthetic lethality in vitro (A) Heatmap showing RT-qPCR analysis of lipid metabolism targets in BON-1 cells treated with mTOR inhibitors (Torin: Torin-1; Evero: everolimus) with or without PIKfyve antagonists. Statistical analysis using two-way ANOVA. (B) Immunoblot analysis of lipid metabolism proteins in QGP-1 cells treated as (A). GAPDH, loading control. (C) Everolimus IC 50 curves in QGP-1 cells with or without CRISPRi-mediated PIKFYVE knockdown. Inset shows IC 50 values. (D) Confluence assay showing the efficacy of everolimus (5 μM) upon PIKFYVE knockdown in QGP-1 cells. Data shown are mean ± SD ( n = 3). Two-way ANOVA. (E and F) 3D synergy plots and heatmaps for QGP-1 cells treated with everolimus and apilimod or ESK981. Red peaks indicate synergy, with the average synergy score shown. (G and H) Confluence assay showing synergistic effect of apilimod (1 μM) or ESK981 (250 nM) combined with everolimus (5 μM). Data presented as mean ± SD ( n = 3). Statistical analysis using two-way ANOVA. (I) 3D synergy plots and heatmaps for apilimod and everolimus with rescue by DMSO, Ferrostatin-1 (1 μM), or Z-VAD-FMK (10 μM). (J) Confluence assay showing synergistic effect of apilimod (1 μM) and everolimus (5 μM) rescued with DMSO, Ferrostatin-1 (1 μM), or Z-VAD-FMK (10 μM). Data presented as mean ± SD ( n = 3). Statistical analysis using two-way ANOVA.

Article Snippet: BON-1 was purchased from Creative Biolabs.

Techniques: Inhibition, In Vitro, Quantitative RT-PCR, Western Blot, Control, Knockdown

Journal: Cell Reports Medicine

Article Title: Targeting the ferritinophagy-lysosome axis as a therapeutic vulnerability in gastroenteropancreatic neuroendocrine tumors

doi: 10.1016/j.xcrm.2026.102695

Figure Lengend Snippet: Combinatorial targeting of mTOR and PIKfyve exerts synergistic effects in vivo in GEP-NETs (A) Immunoblot of QGP-1 and BON-1 CDX tumors after 5 days (PD5) of treatment with vehicle, ESK981 (30 mg/kg), everolimus (2.5 mg/kg), or the combination, showing cleaved PARP (c-PARP); GAPDH, loading control. (B) Schematic of orthotopic pancreatic neuroendocrine tumor model (QGP-1 or BON-1) and treatment regimens. (C) Tumor-to-body weight ratio of pancreas from QGP-1 orthotopic model. (D) Percentage normal pancreas area from the study in (C). Data presented as mean ± SEM. p values calculated using one-way ANOVA. (E) Bioluminescence imaging (BLI) of QGP-1 orthotopic tumors across treatment groups. (F) Kaplan-Meier survival curves of QGP-1 tumor-bearing mice. Data expressed as means ± SEM. p values calculated using two-way repeated-measures ANOVA. (G) Percentage body weight changes of QGP-1 tumor-bearing mice. Data presented as mean ± SEM. (H) BLI of BON-1 orthotopic tumors. Data presented as mean ± SEM. (I) Percentage change in BLI signal on day 28 compared to day 0 for BON-1 orthotopic model shown in (H). (J) Kaplan-Meier survival curves of BON-1 tumor-bearing mice. Data expressed as means ± SEM. p values calculated using two-way repeated-measures ANOVA.

Article Snippet: BON-1 was purchased from Creative Biolabs.

Techniques: In Vivo, Western Blot, Control, Imaging

Journal: Cell Reports Medicine

Article Title: Targeting the ferritinophagy-lysosome axis as a therapeutic vulnerability in gastroenteropancreatic neuroendocrine tumors

doi: 10.1016/j.xcrm.2026.102695

Figure Lengend Snippet: Kinome-wide CRISPR knockout screen identifies PIKfyve as a druggable target in GEP-NETs (A) Schematic of the CRISPR screening workflow in BON-1-DsRed-IRES-GFP-p62 cells. (B) Gene enrichment rank plot from kinome CRISPR knockout screens (specified in ). Cutoff was set at fold change < −0.3. Red genes are related to the phosphatidylinositol metabolic pathway, while blue genes are related to the mTOR pathway. (C) Pathway enrichment analysis of essential genes from CRISPR screening data in (B) using PANTHER 19.0. Targets with a fold change < −0.3 were considered candidate genes, and pathways with a p value <0.05 and hit targets ≥3 were displayed in the plot. Pathways labeled in red are related to phosphatidylinositol metabolism. (D) Schematic of PI(3,5)P2 biosynthesis regulated by VPS34 ( PIK3C3 ) and PIKfyve. (E) Cell confluence of BON-1 and QGP-1 cells after siRNA-mediated knockdown of PIK3C3 , PIKFYVE , or non-targeting control (siNC). Data are presented as mean ± SD ( n = 4 biological replicates). Two-way ANOVA. (F) Crystal violet staining showing long-term growth inhibition by PIKfyve inhibitor apilimod and mTOR inhibitor (Torin-1 or everolimus) in indicated GEP-NETs cell lines and HPNE cells. (G) Compilation of IC 50 values for everolimus, apilimod, and ESK981 in GEP-NET and HPNE cells. (H and I) Dose-response proliferation curves of everolimus (H) and apilimod (I) in indicated GEP-NET cell lines and HPNE cells. Data are presented as mean ± SD (at least three biological replicates).

Article Snippet: BON-1 , Creative Biolabs , RRID: CVCL_3985.

Techniques: CRISPR, Knock-Out, Labeling, Knockdown, Control, Staining, Inhibition

Journal: Cell Reports Medicine

Article Title: Targeting the ferritinophagy-lysosome axis as a therapeutic vulnerability in gastroenteropancreatic neuroendocrine tumors

doi: 10.1016/j.xcrm.2026.102695

Figure Lengend Snippet: PIKfyve is overexpressed in GEP-NETs and serves as a therapeutic target (A) Representative PIKfyve IHC staining in human normal colon tissue, colon neuroendocrine tumor, and colon adenocarcinoma samples. (B) Quantification of PIKfyve H-score from tissue microarray of human normal GEP tissue, GEP-NETs, and GEP adenocarcinoma (specified in ). Statistics were performed using one-way ANOVA. (C) Immunoblot analysis of PIKfyve and autophagy markers (p62 and LC3A/B) in GEP-NET cell lines following CRISPRi-mediated PIKFYVE knockdown. Vinculin served as the loading control. (D) Cell confluence of GEP-NET cell lines with CRISPRi-mediated PIKFYVE (sg PIKFYVE or sg Pikfyve ) knockdown or control (sgNC). Data are presented as mean ± SD ( n = 4 biological replicates). Two-way ANOVA. (E) Immunoblot analysis of autophagy markers in QGP-1 and BON-1 cells following PIKfyve inhibitors (apilimod or ESK981) or PIKfyve degrader (PIK5-33d) treatment for 8 or 24 h. GAPDH was used as a loading control. (F) Average tumor volumes of QGP-1 subcutaneous-cell-line-derived (CDX) model for vehicle ( n = 9) or ESK981 (30 mg/kg, n = 10) treatment. Mean ± SEM. Two-way ANOVA. s.c., subcutaneous. (G) Spider plot displaying individual tumor volumes from vehicle or ESK981 treatment groups in QGP-1 subcutaneous CDX study. (H) Individual tumor weights of QGP-1 subcutaneous CDX model tumors at study endpoint. Unpaired two-tailed t test. (I) Percent body weight change of QGP-1 subcutaneous CDX model tumors from vehicle or ESK981 treatment groups. (J) Average tumor volumes of BON-1 subcutaneous CDX model following vehicle ( n = 17) or ESK981 (30 mg/kg, n = 15) treatment. Mean ± SEM. Two-way ANOVA. (K) Spider plot displaying individual tumor volumes from vehicle or ESK981 treatment groups in BON-1 subcutaneous CDX study. (L) Individual tumor weights of BON-1 subcutaneous CDX model tumors at study endpoint. Unpaired two-tailed t test. (M) Percent body weight change of BON-1 subcutaneous CDX model tumors from vehicle or ESK981 treatment groups.

Article Snippet: BON-1 , Creative Biolabs , RRID: CVCL_3985.

Techniques: Immunohistochemistry, Microarray, Western Blot, Knockdown, Control, Derivative Assay, Two Tailed Test

Journal: Cell Reports Medicine

Article Title: Targeting the ferritinophagy-lysosome axis as a therapeutic vulnerability in gastroenteropancreatic neuroendocrine tumors

doi: 10.1016/j.xcrm.2026.102695

Figure Lengend Snippet: PIKfyve mediates lipid homeostasis in GEP-NETs (A) Pathway enrichment analysis of RNA-seq from QGP-1 cells following CRISPRi-mediated PIKFYVE knockdown. (B) GSEA of fatty acid metabolism, cholesterol homeostasis, and mTORC1 signaling after CRISPRi-mediated PIKFYVE knockdown in QGP-1 cells. (C) Volcano plot of differentially expressed genes highlighting fatty acid metabolism (violet), cholesterol homeostasis (rose), or mTORC1 signaling pathways (orange). (D) Pathway enrichment analysis of RNA-seq from QGP-1 cells treated with PIKfyve inhibitor apilimod (1 μM, 8 h). (E) GSEA of fatty acid metabolism, cholesterol homeostasis, and mTORC1 signaling following apilimod treatment. (F) Volcano plots of differentially expressed genes from QGP-1 cells after apilimod treatment, highlighting fatty acid metabolism (violet), cholesterol homeostasis (rose), or mTORC1 signaling pathways (orange). (G) Schematic illustrating SREBP- and mTOR-dependent regulation of fatty acid cholesterol biosynthesis. (H) Immunoblot showing PIKfyve, premature SREBP1 (p), mature SREBP1 (m), FASN, and SCD expression in QGP-1 and BON-1 cells following genetic or pharmacological PIKfyve inhibition (inhibitors: apilimod, ESK981; degrader: PIK5-33d. 8-h treatment). GAPDH was used as a loading control. (I) LAMP1 and filipin (cholesterol probe) staining showing lysosomal cholesterol accumulation after apilimod or ESK981 treatment for 24 h at 1 μM. Scale bars: 5 μm. (J and K) Synergy analyses of apilimod and the SCD inhibitor (CAY10566) in QGP-1 (J) and STC-1 (K) cells, shown as dose-response heatmaps and 3D synergy plots.

Article Snippet: BON-1 , Creative Biolabs , RRID: CVCL_3985.

Techniques: RNA Sequencing, Knockdown, Protein-Protein interactions, Western Blot, Expressing, Inhibition, Control, Staining

Journal: Cell Reports Medicine

Article Title: Targeting the ferritinophagy-lysosome axis as a therapeutic vulnerability in gastroenteropancreatic neuroendocrine tumors

doi: 10.1016/j.xcrm.2026.102695

Figure Lengend Snippet: Inhibition of the mTOR pathway suppresses the SREBP1 pathway and triggers ferritinophagy (A) Pathway enrichment analysis of RNA-seq in QGP-1 cells treated with Torin-1 (0.1 μM) for 8 h (B) Volcano plot of differentially expressed proteins from whole-cell proteomics of QGP-1 cells after Torin-1 treatment (0.1 μM, 24 h) (specified in ), highlighting fatty acid metabolism, cholesterol homeostasis, and mTORC1 signaling pathways. (C) Immunoblot analysis of mTOR pathway and lipid metabolism proteins in BON-1 and QGP-1 cells after 24 h of the indicated treatment. GAPDH was used as a loading control. (D) Immunoblot showing autophagy-related proteins in BON-1 and QGP-1 cells following Torin-1 or everolimus treatment for 24 h. GAPDH was used as a loading control. (E) Tandem fluorescent reporter assay assessing autophagic flux in QGP-1 and BON-1 cells treated with Torin-1 (0.1 μM) or everolimus (5 μM) for 24 h. One-way ANOVA. (F) Schematic of TMEM192-labeled cells for lysosome isolation and proteomic analysis. (G) Volcano plot of differentially expressed lysosomal proteins of QGP-1 TMEM192 cells treated with Torin-1 (0.1 μM, 24 h) (specified in ), highlighting proteins in mTORC1 signaling (orange) and ferritinophagy (green). (H) Immunoblot validating increased ferritin levels (FTL and FTH1) in QGP-1 TMEM192 cells following Torin-1 treatment for 24 h. GAPDH was used as a loading control for whole-cell lysates, while LAMP1 served as a loading control for lysosomal samples. (I and J) Immunoblot analysis of ferritin and transferrin levels in BON-1 and QGP-1 cells treated with mTOR inhibitor (Torin-1, everolimus) for 24 h or siRNA-mediated knockdown of mTOR. GAPDH and β-actin were used as loading controls. (K) Representative image of FerroOrange staining in QGP-1 cells with the indicated compounds. Torin-1 was used at 100 nM, and Ferrous ammonium sulfate (FAS) was used as a positive control (50 μM). Scale bars: 20 μm. (L) Intracellular iron levels measured using FerroOrange dye via flow cytometry in QGP-1 cells treated with the indicated compounds. Torin-1 was used at 100 nM, and FAS was used at 100 μM. One-way ANOVA. (M) Confluence assay showing enhanced growth inhibition by everolimus (5 μM) combined with iron chelating agent deferoxamine (DFO, 20 μM). Data are presented as mean ± SD ( n = 3∼4). Two-way ANOVA.

Article Snippet: BON-1 , Creative Biolabs , RRID: CVCL_3985.

Techniques: Inhibition, RNA Sequencing, Protein-Protein interactions, Western Blot, Control, Reporter Assay, Labeling, Isolation, Knockdown, Staining, Positive Control, Flow Cytometry

Journal: Cell Reports Medicine

Article Title: Targeting the ferritinophagy-lysosome axis as a therapeutic vulnerability in gastroenteropancreatic neuroendocrine tumors

doi: 10.1016/j.xcrm.2026.102695

Figure Lengend Snippet: PIKfyve blockade abrogates mTOR-inhibition-induced ferritinophagy (A and B) Autophagic flux in QGP-1 (A) and BON-1 (B) cells treated with DMSO, apilimod (AP), ESK981 (ESK), or combinations with Torin-1 (0.1 μM) or everolimus (5 μM) for 24 h. Statistical analysis using one-way ANOVA. (C) Immunoblot analysis of mTOR signaling and autophagy markers in QGP-1 and BON-1 cells treated with mTOR inhibitors with or without PIKfyve antagonists. GAPDH served as a loading control. (D) Volcano plot of differentially expressed lysosomal proteins in QGP-1 TMEM192 cells treated with or without apilimod (1 μM, 24 h) (specified in ). (E) Pathway enrichment analysis of lysosomal proteomics highlighting ferritinophagy-related pathways. (F) Immunoblot validation of decreased ferritin levels (FTL and FTH1) in QGP-1 TMEM192 cells following the indicated treatment for 24 h. GAPDH served as loading control for whole-cell lysates; LAMP1 served as loading control for lysosomal samples. (G and H) Intracellular iron levels in QGP-1 cells assessed by FerroOrange staining (G) and flow cytometry (H). FAS served as a positive control; deferoxamine (DFO) served as a negative control. Scale bars: 20 μm. Statistical analysis using two-way ANOVA. (I) Immunoblot analysis of SDHB following changes in bioavailable iron after lysosomal or PIKfyve inhibition with or without ammonium ferric citrate (FAC) supplementation in indicated cells. β-actin served as the loading control. (J) Confluence assay showing FAC (100 μg/mL)-mediated rescue of cell growth after apilimod treatment. All conditions were supplemented with 1 μM Ferrostatin-1 to prevent the deleterious effects of excess free iron. Data presented as mean ± SD ( n = 3). Statistical analysis using two-way ANOVA.

Article Snippet: BON-1 , Creative Biolabs , RRID: CVCL_3985.

Techniques: Inhibition, Western Blot, Control, Biomarker Discovery, Staining, Flow Cytometry, Positive Control, Negative Control

Journal: Cell Reports Medicine

Article Title: Targeting the ferritinophagy-lysosome axis as a therapeutic vulnerability in gastroenteropancreatic neuroendocrine tumors

doi: 10.1016/j.xcrm.2026.102695

Figure Lengend Snippet: Dual inhibition of mTOR and PIKfyve triggers synthetic lethality in vitro (A) Heatmap showing RT-qPCR analysis of lipid metabolism targets in BON-1 cells treated with mTOR inhibitors (Torin: Torin-1; Evero: everolimus) with or without PIKfyve antagonists. Statistical analysis using two-way ANOVA. (B) Immunoblot analysis of lipid metabolism proteins in QGP-1 cells treated as (A). GAPDH, loading control. (C) Everolimus IC 50 curves in QGP-1 cells with or without CRISPRi-mediated PIKFYVE knockdown. Inset shows IC 50 values. (D) Confluence assay showing the efficacy of everolimus (5 μM) upon PIKFYVE knockdown in QGP-1 cells. Data shown are mean ± SD ( n = 3). Two-way ANOVA. (E and F) 3D synergy plots and heatmaps for QGP-1 cells treated with everolimus and apilimod or ESK981. Red peaks indicate synergy, with the average synergy score shown. (G and H) Confluence assay showing synergistic effect of apilimod (1 μM) or ESK981 (250 nM) combined with everolimus (5 μM). Data presented as mean ± SD ( n = 3). Statistical analysis using two-way ANOVA. (I) 3D synergy plots and heatmaps for apilimod and everolimus with rescue by DMSO, Ferrostatin-1 (1 μM), or Z-VAD-FMK (10 μM). (J) Confluence assay showing synergistic effect of apilimod (1 μM) and everolimus (5 μM) rescued with DMSO, Ferrostatin-1 (1 μM), or Z-VAD-FMK (10 μM). Data presented as mean ± SD ( n = 3). Statistical analysis using two-way ANOVA.

Article Snippet: BON-1 , Creative Biolabs , RRID: CVCL_3985.

Techniques: Inhibition, In Vitro, Quantitative RT-PCR, Western Blot, Control, Knockdown

Journal: Cell Reports Medicine

Article Title: Targeting the ferritinophagy-lysosome axis as a therapeutic vulnerability in gastroenteropancreatic neuroendocrine tumors

doi: 10.1016/j.xcrm.2026.102695

Figure Lengend Snippet: Combinatorial targeting of mTOR and PIKfyve exerts synergistic effects in vivo in GEP-NETs (A) Immunoblot of QGP-1 and BON-1 CDX tumors after 5 days (PD5) of treatment with vehicle, ESK981 (30 mg/kg), everolimus (2.5 mg/kg), or the combination, showing cleaved PARP (c-PARP); GAPDH, loading control. (B) Schematic of orthotopic pancreatic neuroendocrine tumor model (QGP-1 or BON-1) and treatment regimens. (C) Tumor-to-body weight ratio of pancreas from QGP-1 orthotopic model. (D) Percentage normal pancreas area from the study in (C). Data presented as mean ± SEM. p values calculated using one-way ANOVA. (E) Bioluminescence imaging (BLI) of QGP-1 orthotopic tumors across treatment groups. (F) Kaplan-Meier survival curves of QGP-1 tumor-bearing mice. Data expressed as means ± SEM. p values calculated using two-way repeated-measures ANOVA. (G) Percentage body weight changes of QGP-1 tumor-bearing mice. Data presented as mean ± SEM. (H) BLI of BON-1 orthotopic tumors. Data presented as mean ± SEM. (I) Percentage change in BLI signal on day 28 compared to day 0 for BON-1 orthotopic model shown in (H). (J) Kaplan-Meier survival curves of BON-1 tumor-bearing mice. Data expressed as means ± SEM. p values calculated using two-way repeated-measures ANOVA.

Article Snippet: BON-1 , Creative Biolabs , RRID: CVCL_3985.

Techniques: In Vivo, Western Blot, Control, Imaging