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Asterand Inc ilc cell lines sum44pe (sum44)
A Phase contrast light microscopy images of parental and tamoxifen-resistant ILC cells with magnified images in the inserts. B Growth kinetics of parental and tamoxifen-resistant MB134 (upper panel) and <t>SUM44</t> (lower panel) cell lines. Fold change in growth normalized to day 0 at each time point over 72 h. C Dose response to tamoxifen in MB134 and MB134TAMR cells (upper panel) and SUM44 and SUM44TAMR cells (lower panel). Overnight cultures of exponentially growing cells were treated with vehicle or drugs for 5 days. IC 50 for TAM was calculated using GraphPad Prism 10. D Transwell Boyden chamber assays comparing the migratory capabilities of parental vs. TAMR, MB134 (upper panel) and SUM44 (lower panel) cell lines. The area covered by migrated cells was quantified in the bar diagram. E Representative image of western blot and densitometry analysis of ERα and HER2 levels in parental vs. TAMR ILC cells. GAPDH was used as a loading control. Representative of three independent experiments is presented in the figures. Statistical differences between groups were evaluated using Student’s t -test. Significance levels are indicated as follows: ** p ≤ 0.01, * p < 0.05. P Parental, TAMR Tamoxifen Resistant.
Ilc Cell Lines Sum44pe (Sum44), supplied by Asterand Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/sum44pe/sum44pe/pmc12254388-305-8-10
Average 90 stars, based on 1 article reviews
ilc cell lines sum44pe (sum44) - by Bioz Stars, 2026-10
90/100 stars

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1) Product Images from "Invasive lobular carcinoma: integrated multi-omics analysis reveals silencing of Argininosuccinate synthase and upregulation of nucleotide biosynthesis in tamoxifen resistance"

Article Title: Invasive lobular carcinoma: integrated multi-omics analysis reveals silencing of Argininosuccinate synthase and upregulation of nucleotide biosynthesis in tamoxifen resistance

Journal: Cell Death & Disease

doi: 10.1038/s41419-025-07788-6

A Phase contrast light microscopy images of parental and tamoxifen-resistant ILC cells with magnified images in the inserts. B Growth kinetics of parental and tamoxifen-resistant MB134 (upper panel) and SUM44 (lower panel) cell lines. Fold change in growth normalized to day 0 at each time point over 72 h. C Dose response to tamoxifen in MB134 and MB134TAMR cells (upper panel) and SUM44 and SUM44TAMR cells (lower panel). Overnight cultures of exponentially growing cells were treated with vehicle or drugs for 5 days. IC 50 for TAM was calculated using GraphPad Prism 10. D Transwell Boyden chamber assays comparing the migratory capabilities of parental vs. TAMR, MB134 (upper panel) and SUM44 (lower panel) cell lines. The area covered by migrated cells was quantified in the bar diagram. E Representative image of western blot and densitometry analysis of ERα and HER2 levels in parental vs. TAMR ILC cells. GAPDH was used as a loading control. Representative of three independent experiments is presented in the figures. Statistical differences between groups were evaluated using Student’s t -test. Significance levels are indicated as follows: ** p ≤ 0.01, * p < 0.05. P Parental, TAMR Tamoxifen Resistant.
Figure Legend Snippet: A Phase contrast light microscopy images of parental and tamoxifen-resistant ILC cells with magnified images in the inserts. B Growth kinetics of parental and tamoxifen-resistant MB134 (upper panel) and SUM44 (lower panel) cell lines. Fold change in growth normalized to day 0 at each time point over 72 h. C Dose response to tamoxifen in MB134 and MB134TAMR cells (upper panel) and SUM44 and SUM44TAMR cells (lower panel). Overnight cultures of exponentially growing cells were treated with vehicle or drugs for 5 days. IC 50 for TAM was calculated using GraphPad Prism 10. D Transwell Boyden chamber assays comparing the migratory capabilities of parental vs. TAMR, MB134 (upper panel) and SUM44 (lower panel) cell lines. The area covered by migrated cells was quantified in the bar diagram. E Representative image of western blot and densitometry analysis of ERα and HER2 levels in parental vs. TAMR ILC cells. GAPDH was used as a loading control. Representative of three independent experiments is presented in the figures. Statistical differences between groups were evaluated using Student’s t -test. Significance levels are indicated as follows: ** p ≤ 0.01, * p < 0.05. P Parental, TAMR Tamoxifen Resistant.

Techniques Used: Light Microscopy, Western Blot, Control

A Partial Least Squares Discriminant Analysis (PLS-DA) showing metabolic differences between parental and tamoxifen-resistant SUM44 cell pair. B Variable Importance in Projection (VIP) plot highlighting the top 15 metabolites driving the separation between the parental and resistant SUM44 cell pair. C PLS-DA showing metabolic differences between parental and tamoxifen-resistant MB134 cell pair. D VIP plot highlighting the top 15 metabolites driving the separation between the parental and resistant MB134 cell pair. E Overlap of altered pathways in parental vs. tamoxifen-resistant SUM44 and MB134 pairs, represented by impact scores. F Venn diagram illustrating the number of shared and unique pathways altered in SUM44 and MB134 cell pairs, indicating common metabolic changes associated with tamoxifen resistance. G Significantly deregulated KEGG pathways in parental vs. tamoxifen-resistant MB134 cells and H SUM44 cells as determined by RNA sequencing analysis of cell pairs in quadruplicate. I Venn diagram showing the overlap of 52 deregulated pathways between parental and tamoxifen-resistant pairs of SUM44 and MB134 cell lines. J Venn diagram showing the overlap of metabolomic and transcriptomic data, identifying three mutually deregulated pathways in SUM44 and MB134TAMR cells. K Venn diagram showing the overlap of 15 genes within the three mutually deregulated pathways. L Gene-metabolite interaction map illustrating interactions between three deregulated pathways. Circles denote genes and squares denote metabolites.
Figure Legend Snippet: A Partial Least Squares Discriminant Analysis (PLS-DA) showing metabolic differences between parental and tamoxifen-resistant SUM44 cell pair. B Variable Importance in Projection (VIP) plot highlighting the top 15 metabolites driving the separation between the parental and resistant SUM44 cell pair. C PLS-DA showing metabolic differences between parental and tamoxifen-resistant MB134 cell pair. D VIP plot highlighting the top 15 metabolites driving the separation between the parental and resistant MB134 cell pair. E Overlap of altered pathways in parental vs. tamoxifen-resistant SUM44 and MB134 pairs, represented by impact scores. F Venn diagram illustrating the number of shared and unique pathways altered in SUM44 and MB134 cell pairs, indicating common metabolic changes associated with tamoxifen resistance. G Significantly deregulated KEGG pathways in parental vs. tamoxifen-resistant MB134 cells and H SUM44 cells as determined by RNA sequencing analysis of cell pairs in quadruplicate. I Venn diagram showing the overlap of 52 deregulated pathways between parental and tamoxifen-resistant pairs of SUM44 and MB134 cell lines. J Venn diagram showing the overlap of metabolomic and transcriptomic data, identifying three mutually deregulated pathways in SUM44 and MB134TAMR cells. K Venn diagram showing the overlap of 15 genes within the three mutually deregulated pathways. L Gene-metabolite interaction map illustrating interactions between three deregulated pathways. Circles denote genes and squares denote metabolites.

Techniques Used: RNA Sequencing

A Comparative expression of ASS1 in parental (P) and tamoxifen-resistant (TAMR) ILC cell lines as determined by change in Fragments Per Kilobase of transcript per Million mapped reads (FPKM) obtained from RNA-seq data. B qRT-PCR analysis showing ASS1 expression, and C western blot and densitometry analysis showing ASS1 protein levels in MB134 and SUM44 cells, their respective tamoxifen-resistant and LTED derivatives (LTEDA and LTEDB). D Schematic diagram showing the CpG island in the ASS1 promoter. E Analysis of the ASS1 promoter region using MSPCR showing amplification of 150 bp methylated DNA in TAMR and LTED derivatives of ILC cell lines and 147 bp unmethylated DNA in parental ILC cell lines. F qRT-PCR analysis showing ASS1 expression in MB134TAMR and SUM44TAMR cells that are either left untreated (UT) or treated with 5 μM 5-aza-2′-deoxycytidine (dAzaC) for 120 h. G Western blot and densitometry analysis showing ASS1 protein levels in MB134TAMR and SUM44TAMR cells after 120 h of dAzaC treatment ( n = 2 experiments). Error bars represent the standard deviation of triplicates. Significance levels are indicated as follows: **** p < 0.0001 *** p ≤ 0.001, ** p ≤ 0.01, * p < 0.05.
Figure Legend Snippet: A Comparative expression of ASS1 in parental (P) and tamoxifen-resistant (TAMR) ILC cell lines as determined by change in Fragments Per Kilobase of transcript per Million mapped reads (FPKM) obtained from RNA-seq data. B qRT-PCR analysis showing ASS1 expression, and C western blot and densitometry analysis showing ASS1 protein levels in MB134 and SUM44 cells, their respective tamoxifen-resistant and LTED derivatives (LTEDA and LTEDB). D Schematic diagram showing the CpG island in the ASS1 promoter. E Analysis of the ASS1 promoter region using MSPCR showing amplification of 150 bp methylated DNA in TAMR and LTED derivatives of ILC cell lines and 147 bp unmethylated DNA in parental ILC cell lines. F qRT-PCR analysis showing ASS1 expression in MB134TAMR and SUM44TAMR cells that are either left untreated (UT) or treated with 5 μM 5-aza-2′-deoxycytidine (dAzaC) for 120 h. G Western blot and densitometry analysis showing ASS1 protein levels in MB134TAMR and SUM44TAMR cells after 120 h of dAzaC treatment ( n = 2 experiments). Error bars represent the standard deviation of triplicates. Significance levels are indicated as follows: **** p < 0.0001 *** p ≤ 0.001, ** p ≤ 0.01, * p < 0.05.

Techniques Used: Expressing, RNA Sequencing, Quantitative RT-PCR, Western Blot, Amplification, Methylation, Standard Deviation

MB134 and SUM44 cells were transduced with plasmids expressing shRNA targeting ASS1 or the corresponding empty vector to produce shASS1 and control (pLKO) cell lines. A qRT-PCR analysis showing ASS1 expression, and B Western blot and densitometry analysis of ASS1 protein levels in ASS1 knockdown ( shASS1 ) vs. pLKO (control) MB134 and SUM44 cell lines. C Dose response to tamoxifen and IC 50 of tamoxifen in ASS1 knockdown ( shASS1 ) vs. pLKO (control) MB134 (left panel) and SUM44 cell lines (right panel), assessed using the automated cell counting method. D Comparison of growth kinetics of shASS1 vs. pLKO derivatives of MB134 (left panel) and SUM44 cell lines (right panel). Fold change in growth is normalized to day 0 at each time point over 72 h. E Overall survival analysis of all breast cancer patients in relation to ASS1 expression using the METABRIC dataset. F Overall survival analysis of ILC patients who received endocrine therapy in relation to ASS1 expression using the METABRIC dataset. G Overall survival analysis of breast cancer patients who received endocrine therapy in relation to ASS1 expression using the K–M plotter dataset. **** p < 0.0001, *** p ≤ 0.001, ** P ≤ 0.01, * p < 0.05.
Figure Legend Snippet: MB134 and SUM44 cells were transduced with plasmids expressing shRNA targeting ASS1 or the corresponding empty vector to produce shASS1 and control (pLKO) cell lines. A qRT-PCR analysis showing ASS1 expression, and B Western blot and densitometry analysis of ASS1 protein levels in ASS1 knockdown ( shASS1 ) vs. pLKO (control) MB134 and SUM44 cell lines. C Dose response to tamoxifen and IC 50 of tamoxifen in ASS1 knockdown ( shASS1 ) vs. pLKO (control) MB134 (left panel) and SUM44 cell lines (right panel), assessed using the automated cell counting method. D Comparison of growth kinetics of shASS1 vs. pLKO derivatives of MB134 (left panel) and SUM44 cell lines (right panel). Fold change in growth is normalized to day 0 at each time point over 72 h. E Overall survival analysis of all breast cancer patients in relation to ASS1 expression using the METABRIC dataset. F Overall survival analysis of ILC patients who received endocrine therapy in relation to ASS1 expression using the METABRIC dataset. G Overall survival analysis of breast cancer patients who received endocrine therapy in relation to ASS1 expression using the K–M plotter dataset. **** p < 0.0001, *** p ≤ 0.001, ** P ≤ 0.01, * p < 0.05.

Techniques Used: Transduction, Expressing, shRNA, Plasmid Preparation, Control, Quantitative RT-PCR, Western Blot, Knockdown, Cell Counting, Comparison

A Significantly enriched metabolic pathways in shASS1 vs. pLKO derivatives of MB134 and B SUM44 cell lines. C Changes in the levels of purine (dGMP, dGDP, GDP, GMP, IMP) and D pyrimidine (UDP, UMP, and CMP) nucleotides in parental (P) vs. tamoxifen-resistant (TAMR) MB134 cells. E Changes in the levels of purine (ATP and ADP, dGTP, GDP) and F pyrimidine (Cytosine, UMP) nucleotides in parental (P) vs. tamoxifen-resistant (TAMR) SUM44 cells. G Expression levels of PRPS1 , H PAICS , and I DHODH in MB134 and SUM44 cell pairs as analyzed from RNA-seq data. (FPKM: Fragments Per Kilobase Per Million reads). J Representative picture of pCAD S1859 (pCAD) levels in parental and TAMR cells and K in control (pLKO) vs. shASS1 derivatives of MB134 and SUM44 cell pairs. Bar diagrams show the average of more than one independent experiment ( n = 2). **** p < 0.0001, *** p ≤ 0.001, ** P ≤ 0.01, * p < 0.05.
Figure Legend Snippet: A Significantly enriched metabolic pathways in shASS1 vs. pLKO derivatives of MB134 and B SUM44 cell lines. C Changes in the levels of purine (dGMP, dGDP, GDP, GMP, IMP) and D pyrimidine (UDP, UMP, and CMP) nucleotides in parental (P) vs. tamoxifen-resistant (TAMR) MB134 cells. E Changes in the levels of purine (ATP and ADP, dGTP, GDP) and F pyrimidine (Cytosine, UMP) nucleotides in parental (P) vs. tamoxifen-resistant (TAMR) SUM44 cells. G Expression levels of PRPS1 , H PAICS , and I DHODH in MB134 and SUM44 cell pairs as analyzed from RNA-seq data. (FPKM: Fragments Per Kilobase Per Million reads). J Representative picture of pCAD S1859 (pCAD) levels in parental and TAMR cells and K in control (pLKO) vs. shASS1 derivatives of MB134 and SUM44 cell pairs. Bar diagrams show the average of more than one independent experiment ( n = 2). **** p < 0.0001, *** p ≤ 0.001, ** P ≤ 0.01, * p < 0.05.

Techniques Used: Expressing, RNA Sequencing, Control

Related Articles

Multiple Displacement Amplification:

Article Title: Co-regulator activity of Mediator of DNA Damage Checkpoint 1 (MDC1) is associated with DNA repair dysfunction and PARP inhibitor sensitivity in lobular carcinoma of the breast
Article Snippet: .. MDA MB 134VI (MM134; ATCC HTB-23) and SUM44PE (44PE; Asterand/BioIVT) were maintained as described ( ). .. HCC1428 (ATCC CRL-2327), MCF7 (Rae Lab, U. Michigan) and T47D (Sartorius Lab, U. Colorado) were maintained as described ( ).

Article Title: Loss of E-cadherin Enhances IGF1–IGF1R Pathway Activation and Sensitizes Breast Cancers to Anti-IGF1R/InsR Inhibitors
Article Snippet: .. MCF-7 (ATCC; DMEM+10% FBS [06/29/16]), T47D 129 (ATCC; RPMI+10% FBS [02/08/17]), ZR75.1 (ATCC; RPMI+10% FBS [10/13/16]), MDA-MB-130 231 (ATCC; DMEM+10% FBS [10/13/16]), MDA-MB-134-VI (ATCC; 50/50 DMEM/L15+10% 131 FBS [02/08/17]), SUM44PE (Asterand; DMEM/F12+2% CSS with 5ug/ml insulin, 1ug/ml 132 hydrocortisone, 5mM ethanolamine, 5ug/ml transferrin, 10nM triodothyronime, and 50nM 133 sodium selenite [02/08/17 – no reference profile exists in database]), and BCK426 (MEM+5% 134 FBS with 1nM insulin and 1x NEAA [10/13/16 – no reference profile exists in database) cells 135 were cultured with indicated media conditions. ..

Article Title: Mediator of DNA Damage Checkpoint 1 (MDC1) Is a Novel Estrogen Receptor Coregulator in Invasive Lobular Carcinoma of the Breast
Article Snippet: .. MDA MB 134VI (MM134; ATCC HTB-23, RRID:CVCL_0617) and SUM44PE (Asterand/BioIVT; RRID:CVCL_3424) were maintained as described (10). .. MDA MB 330 (MM330; ATCC HTB-127, RRID:CVCL_0619), HCC1428 (ATCC CRL-2327, RRID:CVCL_YX83), CAMA-1 (RRID:CVCL_1115), and MCF7 (RRID:CVCL_0031) (Rae Lab, U. Michigan) were maintained in DMEM/F12 (Corning #10092CV) þ 10% fetal bovine serum (FBS; Nucleus Biologics #FBS1824).

Article Title: High intra-tumoral stromal content defines Reactive breast cancer as a low-risk breast cancer subtype
Article Snippet: Cell lines were routinely tested for Mycoplasma infection using a MycoTect Kit (Invitrogen). .. MDA-MB-134 (ATCC) and Sum44PE (Asterand) cells were cultured as previously described ( 16 ). ..

Cell Culture:

Article Title: Loss of E-cadherin Enhances IGF1–IGF1R Pathway Activation and Sensitizes Breast Cancers to Anti-IGF1R/InsR Inhibitors
Article Snippet: .. MCF-7 (ATCC; DMEM+10% FBS [06/29/16]), T47D 129 (ATCC; RPMI+10% FBS [02/08/17]), ZR75.1 (ATCC; RPMI+10% FBS [10/13/16]), MDA-MB-130 231 (ATCC; DMEM+10% FBS [10/13/16]), MDA-MB-134-VI (ATCC; 50/50 DMEM/L15+10% 131 FBS [02/08/17]), SUM44PE (Asterand; DMEM/F12+2% CSS with 5ug/ml insulin, 1ug/ml 132 hydrocortisone, 5mM ethanolamine, 5ug/ml transferrin, 10nM triodothyronime, and 50nM 133 sodium selenite [02/08/17 – no reference profile exists in database]), and BCK426 (MEM+5% 134 FBS with 1nM insulin and 1x NEAA [10/13/16 – no reference profile exists in database) cells 135 were cultured with indicated media conditions. ..

Article Title: High intra-tumoral stromal content defines Reactive breast cancer as a low-risk breast cancer subtype
Article Snippet: Cell lines were routinely tested for Mycoplasma infection using a MycoTect Kit (Invitrogen). .. MDA-MB-134 (ATCC) and Sum44PE (Asterand) cells were cultured as previously described ( 16 ). ..

other:

Article Title: Therapeutic combinations using IGF1R pathway inhibitors, and methods to predict anti-IGF1R therapeutic efficacy
Article Snippet: MCF-7 (ATCC; DMEM+10% FBS [Jun.



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A Phase contrast light microscopy images of parental and tamoxifen-resistant ILC cells with magnified images in the inserts. B Growth kinetics of parental and tamoxifen-resistant MB134 (upper panel) and <t>SUM44</t> (lower panel) cell lines. Fold change in growth normalized to day 0 at each time point over 72 h. C Dose response to tamoxifen in MB134 and MB134TAMR cells (upper panel) and SUM44 and SUM44TAMR cells (lower panel). Overnight cultures of exponentially growing cells were treated with vehicle or drugs for 5 days. IC 50 for TAM was calculated using GraphPad Prism 10. D Transwell Boyden chamber assays comparing the migratory capabilities of parental vs. TAMR, MB134 (upper panel) and SUM44 (lower panel) cell lines. The area covered by migrated cells was quantified in the bar diagram. E Representative image of western blot and densitometry analysis of ERα and HER2 levels in parental vs. TAMR ILC cells. GAPDH was used as a loading control. Representative of three independent experiments is presented in the figures. Statistical differences between groups were evaluated using Student’s t -test. Significance levels are indicated as follows: ** p ≤ 0.01, * p < 0.05. P Parental, TAMR Tamoxifen Resistant.
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A Phase contrast light microscopy images of parental and tamoxifen-resistant ILC cells with magnified images in the inserts. B Growth kinetics of parental and tamoxifen-resistant MB134 (upper panel) and <t>SUM44</t> (lower panel) cell lines. Fold change in growth normalized to day 0 at each time point over 72 h. C Dose response to tamoxifen in MB134 and MB134TAMR cells (upper panel) and SUM44 and SUM44TAMR cells (lower panel). Overnight cultures of exponentially growing cells were treated with vehicle or drugs for 5 days. IC 50 for TAM was calculated using GraphPad Prism 10. D Transwell Boyden chamber assays comparing the migratory capabilities of parental vs. TAMR, MB134 (upper panel) and SUM44 (lower panel) cell lines. The area covered by migrated cells was quantified in the bar diagram. E Representative image of western blot and densitometry analysis of ERα and HER2 levels in parental vs. TAMR ILC cells. GAPDH was used as a loading control. Representative of three independent experiments is presented in the figures. Statistical differences between groups were evaluated using Student’s t -test. Significance levels are indicated as follows: ** p ≤ 0.01, * p < 0.05. P Parental, TAMR Tamoxifen Resistant.
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A. Phase contrast light microscopy images of parental and tamoxifen-resistant ILC cells with magnified images in the inserts. B. Growth kinetics of parental and tamoxifen-resistant MB134 (upper panel) and <t>SUM44</t> (lower panel) cell lines. Fold change in growth normalized to day 0 at each time point over 72 hours. C. Dose response to tamoxifen in MB134 and MB134TAMR cells (upper panel) and SUM44 and SUM44TAMR cells (lower panel). Overnight cultures of exponentially growing cells were treated with vehicle or drugs for 5 days. IC 50 for TAM was calculated using GraphPad Prism 10. D. Transwell Boyden chamber assays comparing the migratory capabilities of parental vs. TAMR, MB134 (upper panel) and SUM44 (lower panel) cell lines. Area covered by migrated cells were quantified in the bar diagram. E. Representative image of western blot and densitometry analysis of ERα and HER2 levels in parental vs. TAMR ILC cells. GAPDH was used as loading control. Representative of three independent experiments is presented in the figures. Statistical differences between groups were evaluated using Student’s t-test. Significance levels are indicated as follows: **p <0.001, *p <0.05. P= Parental, TAMR = Tamoxifen Resistant
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A. Phase contrast light microscopy images of parental and tamoxifen-resistant ILC cells with magnified images in the inserts. B. Growth kinetics of parental and tamoxifen-resistant MB134 (upper panel) and <t>SUM44</t> (lower panel) cell lines. Fold change in growth normalized to day 0 at each time point over 72 hours. C. Dose response to tamoxifen in MB134 and MB134TAMR cells (upper panel) and SUM44 and SUM44TAMR cells (lower panel). Overnight cultures of exponentially growing cells were treated with vehicle or drugs for 5 days. IC 50 for TAM was calculated using GraphPad Prism 10. D. Transwell Boyden chamber assays comparing the migratory capabilities of parental vs. TAMR, MB134 (upper panel) and SUM44 (lower panel) cell lines. Area covered by migrated cells were quantified in the bar diagram. E. Representative image of western blot and densitometry analysis of ERα and HER2 levels in parental vs. TAMR ILC cells. GAPDH was used as loading control. Representative of three independent experiments is presented in the figures. Statistical differences between groups were evaluated using Student’s t-test. Significance levels are indicated as follows: **p <0.001, *p <0.05. P= Parental, TAMR = Tamoxifen Resistant
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A. Phase contrast light microscopy images of parental and tamoxifen-resistant ILC cells with magnified images in the inserts. B. Growth kinetics of parental and tamoxifen-resistant MB134 (upper panel) and <t>SUM44</t> (lower panel) cell lines. Fold change in growth normalized to day 0 at each time point over 72 hours. C. Dose response to tamoxifen in MB134 and MB134TAMR cells (upper panel) and SUM44 and SUM44TAMR cells (lower panel). Overnight cultures of exponentially growing cells were treated with vehicle or drugs for 5 days. IC 50 for TAM was calculated using GraphPad Prism 10. D. Transwell Boyden chamber assays comparing the migratory capabilities of parental vs. TAMR, MB134 (upper panel) and SUM44 (lower panel) cell lines. Area covered by migrated cells were quantified in the bar diagram. E. Representative image of western blot and densitometry analysis of ERα and HER2 levels in parental vs. TAMR ILC cells. GAPDH was used as loading control. Representative of three independent experiments is presented in the figures. Statistical differences between groups were evaluated using Student’s t-test. Significance levels are indicated as follows: **p <0.001, *p <0.05. P= Parental, TAMR = Tamoxifen Resistant
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A. Phase contrast light microscopy images of parental and tamoxifen-resistant ILC cells with magnified images in the inserts. B. Growth kinetics of parental and tamoxifen-resistant MB134 (upper panel) and <t>SUM44</t> (lower panel) cell lines. Fold change in growth normalized to day 0 at each time point over 72 hours. C. Dose response to tamoxifen in MB134 and MB134TAMR cells (upper panel) and SUM44 and SUM44TAMR cells (lower panel). Overnight cultures of exponentially growing cells were treated with vehicle or drugs for 5 days. IC 50 for TAM was calculated using GraphPad Prism 10. D. Transwell Boyden chamber assays comparing the migratory capabilities of parental vs. TAMR, MB134 (upper panel) and SUM44 (lower panel) cell lines. Area covered by migrated cells were quantified in the bar diagram. E. Representative image of western blot and densitometry analysis of ERα and HER2 levels in parental vs. TAMR ILC cells. GAPDH was used as loading control. Representative of three independent experiments is presented in the figures. Statistical differences between groups were evaluated using Student’s t-test. Significance levels are indicated as follows: **p <0.001, *p <0.05. P= Parental, TAMR = Tamoxifen Resistant
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Image Search Results


A Phase contrast light microscopy images of parental and tamoxifen-resistant ILC cells with magnified images in the inserts. B Growth kinetics of parental and tamoxifen-resistant MB134 (upper panel) and SUM44 (lower panel) cell lines. Fold change in growth normalized to day 0 at each time point over 72 h. C Dose response to tamoxifen in MB134 and MB134TAMR cells (upper panel) and SUM44 and SUM44TAMR cells (lower panel). Overnight cultures of exponentially growing cells were treated with vehicle or drugs for 5 days. IC 50 for TAM was calculated using GraphPad Prism 10. D Transwell Boyden chamber assays comparing the migratory capabilities of parental vs. TAMR, MB134 (upper panel) and SUM44 (lower panel) cell lines. The area covered by migrated cells was quantified in the bar diagram. E Representative image of western blot and densitometry analysis of ERα and HER2 levels in parental vs. TAMR ILC cells. GAPDH was used as a loading control. Representative of three independent experiments is presented in the figures. Statistical differences between groups were evaluated using Student’s t -test. Significance levels are indicated as follows: ** p ≤ 0.01, * p < 0.05. P Parental, TAMR Tamoxifen Resistant.

Journal: Cell Death & Disease

Article Title: Invasive lobular carcinoma: integrated multi-omics analysis reveals silencing of Argininosuccinate synthase and upregulation of nucleotide biosynthesis in tamoxifen resistance

doi: 10.1038/s41419-025-07788-6

Figure Lengend Snippet: A Phase contrast light microscopy images of parental and tamoxifen-resistant ILC cells with magnified images in the inserts. B Growth kinetics of parental and tamoxifen-resistant MB134 (upper panel) and SUM44 (lower panel) cell lines. Fold change in growth normalized to day 0 at each time point over 72 h. C Dose response to tamoxifen in MB134 and MB134TAMR cells (upper panel) and SUM44 and SUM44TAMR cells (lower panel). Overnight cultures of exponentially growing cells were treated with vehicle or drugs for 5 days. IC 50 for TAM was calculated using GraphPad Prism 10. D Transwell Boyden chamber assays comparing the migratory capabilities of parental vs. TAMR, MB134 (upper panel) and SUM44 (lower panel) cell lines. The area covered by migrated cells was quantified in the bar diagram. E Representative image of western blot and densitometry analysis of ERα and HER2 levels in parental vs. TAMR ILC cells. GAPDH was used as a loading control. Representative of three independent experiments is presented in the figures. Statistical differences between groups were evaluated using Student’s t -test. Significance levels are indicated as follows: ** p ≤ 0.01, * p < 0.05. P Parental, TAMR Tamoxifen Resistant.

Article Snippet: ILC cell lines MDA-MB134-VI (MB134) (ATCC, USA) and SUM44PE (SUM44) (Asterand, USA) were used to develop the tamoxifen-resistant (TAMR) cells by continuously exposing them to 100–500 nM of 4-hydroxy tamoxifen (4-OHT) for over 6 months.

Techniques: Light Microscopy, Western Blot, Control

A Partial Least Squares Discriminant Analysis (PLS-DA) showing metabolic differences between parental and tamoxifen-resistant SUM44 cell pair. B Variable Importance in Projection (VIP) plot highlighting the top 15 metabolites driving the separation between the parental and resistant SUM44 cell pair. C PLS-DA showing metabolic differences between parental and tamoxifen-resistant MB134 cell pair. D VIP plot highlighting the top 15 metabolites driving the separation between the parental and resistant MB134 cell pair. E Overlap of altered pathways in parental vs. tamoxifen-resistant SUM44 and MB134 pairs, represented by impact scores. F Venn diagram illustrating the number of shared and unique pathways altered in SUM44 and MB134 cell pairs, indicating common metabolic changes associated with tamoxifen resistance. G Significantly deregulated KEGG pathways in parental vs. tamoxifen-resistant MB134 cells and H SUM44 cells as determined by RNA sequencing analysis of cell pairs in quadruplicate. I Venn diagram showing the overlap of 52 deregulated pathways between parental and tamoxifen-resistant pairs of SUM44 and MB134 cell lines. J Venn diagram showing the overlap of metabolomic and transcriptomic data, identifying three mutually deregulated pathways in SUM44 and MB134TAMR cells. K Venn diagram showing the overlap of 15 genes within the three mutually deregulated pathways. L Gene-metabolite interaction map illustrating interactions between three deregulated pathways. Circles denote genes and squares denote metabolites.

Journal: Cell Death & Disease

Article Title: Invasive lobular carcinoma: integrated multi-omics analysis reveals silencing of Argininosuccinate synthase and upregulation of nucleotide biosynthesis in tamoxifen resistance

doi: 10.1038/s41419-025-07788-6

Figure Lengend Snippet: A Partial Least Squares Discriminant Analysis (PLS-DA) showing metabolic differences between parental and tamoxifen-resistant SUM44 cell pair. B Variable Importance in Projection (VIP) plot highlighting the top 15 metabolites driving the separation between the parental and resistant SUM44 cell pair. C PLS-DA showing metabolic differences between parental and tamoxifen-resistant MB134 cell pair. D VIP plot highlighting the top 15 metabolites driving the separation between the parental and resistant MB134 cell pair. E Overlap of altered pathways in parental vs. tamoxifen-resistant SUM44 and MB134 pairs, represented by impact scores. F Venn diagram illustrating the number of shared and unique pathways altered in SUM44 and MB134 cell pairs, indicating common metabolic changes associated with tamoxifen resistance. G Significantly deregulated KEGG pathways in parental vs. tamoxifen-resistant MB134 cells and H SUM44 cells as determined by RNA sequencing analysis of cell pairs in quadruplicate. I Venn diagram showing the overlap of 52 deregulated pathways between parental and tamoxifen-resistant pairs of SUM44 and MB134 cell lines. J Venn diagram showing the overlap of metabolomic and transcriptomic data, identifying three mutually deregulated pathways in SUM44 and MB134TAMR cells. K Venn diagram showing the overlap of 15 genes within the three mutually deregulated pathways. L Gene-metabolite interaction map illustrating interactions between three deregulated pathways. Circles denote genes and squares denote metabolites.

Article Snippet: ILC cell lines MDA-MB134-VI (MB134) (ATCC, USA) and SUM44PE (SUM44) (Asterand, USA) were used to develop the tamoxifen-resistant (TAMR) cells by continuously exposing them to 100–500 nM of 4-hydroxy tamoxifen (4-OHT) for over 6 months.

Techniques: RNA Sequencing

A Comparative expression of ASS1 in parental (P) and tamoxifen-resistant (TAMR) ILC cell lines as determined by change in Fragments Per Kilobase of transcript per Million mapped reads (FPKM) obtained from RNA-seq data. B qRT-PCR analysis showing ASS1 expression, and C western blot and densitometry analysis showing ASS1 protein levels in MB134 and SUM44 cells, their respective tamoxifen-resistant and LTED derivatives (LTEDA and LTEDB). D Schematic diagram showing the CpG island in the ASS1 promoter. E Analysis of the ASS1 promoter region using MSPCR showing amplification of 150 bp methylated DNA in TAMR and LTED derivatives of ILC cell lines and 147 bp unmethylated DNA in parental ILC cell lines. F qRT-PCR analysis showing ASS1 expression in MB134TAMR and SUM44TAMR cells that are either left untreated (UT) or treated with 5 μM 5-aza-2′-deoxycytidine (dAzaC) for 120 h. G Western blot and densitometry analysis showing ASS1 protein levels in MB134TAMR and SUM44TAMR cells after 120 h of dAzaC treatment ( n = 2 experiments). Error bars represent the standard deviation of triplicates. Significance levels are indicated as follows: **** p < 0.0001 *** p ≤ 0.001, ** p ≤ 0.01, * p < 0.05.

Journal: Cell Death & Disease

Article Title: Invasive lobular carcinoma: integrated multi-omics analysis reveals silencing of Argininosuccinate synthase and upregulation of nucleotide biosynthesis in tamoxifen resistance

doi: 10.1038/s41419-025-07788-6

Figure Lengend Snippet: A Comparative expression of ASS1 in parental (P) and tamoxifen-resistant (TAMR) ILC cell lines as determined by change in Fragments Per Kilobase of transcript per Million mapped reads (FPKM) obtained from RNA-seq data. B qRT-PCR analysis showing ASS1 expression, and C western blot and densitometry analysis showing ASS1 protein levels in MB134 and SUM44 cells, their respective tamoxifen-resistant and LTED derivatives (LTEDA and LTEDB). D Schematic diagram showing the CpG island in the ASS1 promoter. E Analysis of the ASS1 promoter region using MSPCR showing amplification of 150 bp methylated DNA in TAMR and LTED derivatives of ILC cell lines and 147 bp unmethylated DNA in parental ILC cell lines. F qRT-PCR analysis showing ASS1 expression in MB134TAMR and SUM44TAMR cells that are either left untreated (UT) or treated with 5 μM 5-aza-2′-deoxycytidine (dAzaC) for 120 h. G Western blot and densitometry analysis showing ASS1 protein levels in MB134TAMR and SUM44TAMR cells after 120 h of dAzaC treatment ( n = 2 experiments). Error bars represent the standard deviation of triplicates. Significance levels are indicated as follows: **** p < 0.0001 *** p ≤ 0.001, ** p ≤ 0.01, * p < 0.05.

Article Snippet: ILC cell lines MDA-MB134-VI (MB134) (ATCC, USA) and SUM44PE (SUM44) (Asterand, USA) were used to develop the tamoxifen-resistant (TAMR) cells by continuously exposing them to 100–500 nM of 4-hydroxy tamoxifen (4-OHT) for over 6 months.

Techniques: Expressing, RNA Sequencing, Quantitative RT-PCR, Western Blot, Amplification, Methylation, Standard Deviation

MB134 and SUM44 cells were transduced with plasmids expressing shRNA targeting ASS1 or the corresponding empty vector to produce shASS1 and control (pLKO) cell lines. A qRT-PCR analysis showing ASS1 expression, and B Western blot and densitometry analysis of ASS1 protein levels in ASS1 knockdown ( shASS1 ) vs. pLKO (control) MB134 and SUM44 cell lines. C Dose response to tamoxifen and IC 50 of tamoxifen in ASS1 knockdown ( shASS1 ) vs. pLKO (control) MB134 (left panel) and SUM44 cell lines (right panel), assessed using the automated cell counting method. D Comparison of growth kinetics of shASS1 vs. pLKO derivatives of MB134 (left panel) and SUM44 cell lines (right panel). Fold change in growth is normalized to day 0 at each time point over 72 h. E Overall survival analysis of all breast cancer patients in relation to ASS1 expression using the METABRIC dataset. F Overall survival analysis of ILC patients who received endocrine therapy in relation to ASS1 expression using the METABRIC dataset. G Overall survival analysis of breast cancer patients who received endocrine therapy in relation to ASS1 expression using the K–M plotter dataset. **** p < 0.0001, *** p ≤ 0.001, ** P ≤ 0.01, * p < 0.05.

Journal: Cell Death & Disease

Article Title: Invasive lobular carcinoma: integrated multi-omics analysis reveals silencing of Argininosuccinate synthase and upregulation of nucleotide biosynthesis in tamoxifen resistance

doi: 10.1038/s41419-025-07788-6

Figure Lengend Snippet: MB134 and SUM44 cells were transduced with plasmids expressing shRNA targeting ASS1 or the corresponding empty vector to produce shASS1 and control (pLKO) cell lines. A qRT-PCR analysis showing ASS1 expression, and B Western blot and densitometry analysis of ASS1 protein levels in ASS1 knockdown ( shASS1 ) vs. pLKO (control) MB134 and SUM44 cell lines. C Dose response to tamoxifen and IC 50 of tamoxifen in ASS1 knockdown ( shASS1 ) vs. pLKO (control) MB134 (left panel) and SUM44 cell lines (right panel), assessed using the automated cell counting method. D Comparison of growth kinetics of shASS1 vs. pLKO derivatives of MB134 (left panel) and SUM44 cell lines (right panel). Fold change in growth is normalized to day 0 at each time point over 72 h. E Overall survival analysis of all breast cancer patients in relation to ASS1 expression using the METABRIC dataset. F Overall survival analysis of ILC patients who received endocrine therapy in relation to ASS1 expression using the METABRIC dataset. G Overall survival analysis of breast cancer patients who received endocrine therapy in relation to ASS1 expression using the K–M plotter dataset. **** p < 0.0001, *** p ≤ 0.001, ** P ≤ 0.01, * p < 0.05.

Article Snippet: ILC cell lines MDA-MB134-VI (MB134) (ATCC, USA) and SUM44PE (SUM44) (Asterand, USA) were used to develop the tamoxifen-resistant (TAMR) cells by continuously exposing them to 100–500 nM of 4-hydroxy tamoxifen (4-OHT) for over 6 months.

Techniques: Transduction, Expressing, shRNA, Plasmid Preparation, Control, Quantitative RT-PCR, Western Blot, Knockdown, Cell Counting, Comparison

A Significantly enriched metabolic pathways in shASS1 vs. pLKO derivatives of MB134 and B SUM44 cell lines. C Changes in the levels of purine (dGMP, dGDP, GDP, GMP, IMP) and D pyrimidine (UDP, UMP, and CMP) nucleotides in parental (P) vs. tamoxifen-resistant (TAMR) MB134 cells. E Changes in the levels of purine (ATP and ADP, dGTP, GDP) and F pyrimidine (Cytosine, UMP) nucleotides in parental (P) vs. tamoxifen-resistant (TAMR) SUM44 cells. G Expression levels of PRPS1 , H PAICS , and I DHODH in MB134 and SUM44 cell pairs as analyzed from RNA-seq data. (FPKM: Fragments Per Kilobase Per Million reads). J Representative picture of pCAD S1859 (pCAD) levels in parental and TAMR cells and K in control (pLKO) vs. shASS1 derivatives of MB134 and SUM44 cell pairs. Bar diagrams show the average of more than one independent experiment ( n = 2). **** p < 0.0001, *** p ≤ 0.001, ** P ≤ 0.01, * p < 0.05.

Journal: Cell Death & Disease

Article Title: Invasive lobular carcinoma: integrated multi-omics analysis reveals silencing of Argininosuccinate synthase and upregulation of nucleotide biosynthesis in tamoxifen resistance

doi: 10.1038/s41419-025-07788-6

Figure Lengend Snippet: A Significantly enriched metabolic pathways in shASS1 vs. pLKO derivatives of MB134 and B SUM44 cell lines. C Changes in the levels of purine (dGMP, dGDP, GDP, GMP, IMP) and D pyrimidine (UDP, UMP, and CMP) nucleotides in parental (P) vs. tamoxifen-resistant (TAMR) MB134 cells. E Changes in the levels of purine (ATP and ADP, dGTP, GDP) and F pyrimidine (Cytosine, UMP) nucleotides in parental (P) vs. tamoxifen-resistant (TAMR) SUM44 cells. G Expression levels of PRPS1 , H PAICS , and I DHODH in MB134 and SUM44 cell pairs as analyzed from RNA-seq data. (FPKM: Fragments Per Kilobase Per Million reads). J Representative picture of pCAD S1859 (pCAD) levels in parental and TAMR cells and K in control (pLKO) vs. shASS1 derivatives of MB134 and SUM44 cell pairs. Bar diagrams show the average of more than one independent experiment ( n = 2). **** p < 0.0001, *** p ≤ 0.001, ** P ≤ 0.01, * p < 0.05.

Article Snippet: ILC cell lines MDA-MB134-VI (MB134) (ATCC, USA) and SUM44PE (SUM44) (Asterand, USA) were used to develop the tamoxifen-resistant (TAMR) cells by continuously exposing them to 100–500 nM of 4-hydroxy tamoxifen (4-OHT) for over 6 months.

Techniques: Expressing, RNA Sequencing, Control

A. Phase contrast light microscopy images of parental and tamoxifen-resistant ILC cells with magnified images in the inserts. B. Growth kinetics of parental and tamoxifen-resistant MB134 (upper panel) and SUM44 (lower panel) cell lines. Fold change in growth normalized to day 0 at each time point over 72 hours. C. Dose response to tamoxifen in MB134 and MB134TAMR cells (upper panel) and SUM44 and SUM44TAMR cells (lower panel). Overnight cultures of exponentially growing cells were treated with vehicle or drugs for 5 days. IC 50 for TAM was calculated using GraphPad Prism 10. D. Transwell Boyden chamber assays comparing the migratory capabilities of parental vs. TAMR, MB134 (upper panel) and SUM44 (lower panel) cell lines. Area covered by migrated cells were quantified in the bar diagram. E. Representative image of western blot and densitometry analysis of ERα and HER2 levels in parental vs. TAMR ILC cells. GAPDH was used as loading control. Representative of three independent experiments is presented in the figures. Statistical differences between groups were evaluated using Student’s t-test. Significance levels are indicated as follows: **p <0.001, *p <0.05. P= Parental, TAMR = Tamoxifen Resistant

Journal: bioRxiv

Article Title: Invasive lobular carcinoma integrated multi-omics analysis reveals silencing of Arginosuccinate synthase and upregulation of nucleotide biosynthesis in tamoxifen resistance

doi: 10.1101/2025.01.16.633236

Figure Lengend Snippet: A. Phase contrast light microscopy images of parental and tamoxifen-resistant ILC cells with magnified images in the inserts. B. Growth kinetics of parental and tamoxifen-resistant MB134 (upper panel) and SUM44 (lower panel) cell lines. Fold change in growth normalized to day 0 at each time point over 72 hours. C. Dose response to tamoxifen in MB134 and MB134TAMR cells (upper panel) and SUM44 and SUM44TAMR cells (lower panel). Overnight cultures of exponentially growing cells were treated with vehicle or drugs for 5 days. IC 50 for TAM was calculated using GraphPad Prism 10. D. Transwell Boyden chamber assays comparing the migratory capabilities of parental vs. TAMR, MB134 (upper panel) and SUM44 (lower panel) cell lines. Area covered by migrated cells were quantified in the bar diagram. E. Representative image of western blot and densitometry analysis of ERα and HER2 levels in parental vs. TAMR ILC cells. GAPDH was used as loading control. Representative of three independent experiments is presented in the figures. Statistical differences between groups were evaluated using Student’s t-test. Significance levels are indicated as follows: **p <0.001, *p <0.05. P= Parental, TAMR = Tamoxifen Resistant

Article Snippet: ILC cell lines MDA-MB-134-VI (MB134) (ATCC, USA) and SUM44PE (SUM44) (Asterand, USA) were used to develop the tamoxifen-resistant (TAMR) cells by continuously exposing them to 100 to 500 nM of 4-hydroxy tamoxifen (4-OHT) for over 6 months.

Techniques: Light Microscopy, Western Blot, Control

A. Partial Least Squares Discriminant Analysis (PLS-DA) showing metabolic differences between parental and tamoxifen-resistant SUM44 cell pair. B. Variable Importance in Projection (VIP) plot highlighting the top 15 metabolites driving the separation between parental and resistant SUM44 cell pair. C. PLS-DA showing metabolic differences between parental and tamoxifen-resistant MB134 cell pair. D. VIP plot highlighting the top 15 metabolites driving the separation between parental and resistant MB134 cell pair. E. Overlap of altered pathways in parental vs. tamoxifen-resistant SUM44 and MB134 pairs, represented by impact scores. F. Venn diagram illustrating the number of shared and unique pathways altered in SUM44 and MB134 cell pairs, indicating common metabolic changes associated with tamoxifen resistance. G. Significantly deregulated KEGG pathways in parental vs. tamoxifen-resistant MB134 cells and, H. SUM44 cells as determined by RNA sequencing analysis of cell pairs in quadruplicate. I. Venn diagram showing the overlap of 52 deregulated pathways between parental and tamoxifen-resistant pairs of SUM44 and MB-134 cell lines. J. Venn diagram showing the overlap of metabolomic and transcriptomic data, identifying three mutually deregulated pathways in SUM44 and MB134 TAMR cells. K. Venn diagram showing the overlap of 15 genes within the three mutually deregulated pathways. L. Gene-metabolite interaction map illustrating interactions between three deregulated pathways. Circles represent genes and squares represent metabolites.

Journal: bioRxiv

Article Title: Invasive lobular carcinoma integrated multi-omics analysis reveals silencing of Arginosuccinate synthase and upregulation of nucleotide biosynthesis in tamoxifen resistance

doi: 10.1101/2025.01.16.633236

Figure Lengend Snippet: A. Partial Least Squares Discriminant Analysis (PLS-DA) showing metabolic differences between parental and tamoxifen-resistant SUM44 cell pair. B. Variable Importance in Projection (VIP) plot highlighting the top 15 metabolites driving the separation between parental and resistant SUM44 cell pair. C. PLS-DA showing metabolic differences between parental and tamoxifen-resistant MB134 cell pair. D. VIP plot highlighting the top 15 metabolites driving the separation between parental and resistant MB134 cell pair. E. Overlap of altered pathways in parental vs. tamoxifen-resistant SUM44 and MB134 pairs, represented by impact scores. F. Venn diagram illustrating the number of shared and unique pathways altered in SUM44 and MB134 cell pairs, indicating common metabolic changes associated with tamoxifen resistance. G. Significantly deregulated KEGG pathways in parental vs. tamoxifen-resistant MB134 cells and, H. SUM44 cells as determined by RNA sequencing analysis of cell pairs in quadruplicate. I. Venn diagram showing the overlap of 52 deregulated pathways between parental and tamoxifen-resistant pairs of SUM44 and MB-134 cell lines. J. Venn diagram showing the overlap of metabolomic and transcriptomic data, identifying three mutually deregulated pathways in SUM44 and MB134 TAMR cells. K. Venn diagram showing the overlap of 15 genes within the three mutually deregulated pathways. L. Gene-metabolite interaction map illustrating interactions between three deregulated pathways. Circles represent genes and squares represent metabolites.

Article Snippet: ILC cell lines MDA-MB-134-VI (MB134) (ATCC, USA) and SUM44PE (SUM44) (Asterand, USA) were used to develop the tamoxifen-resistant (TAMR) cells by continuously exposing them to 100 to 500 nM of 4-hydroxy tamoxifen (4-OHT) for over 6 months.

Techniques: RNA Sequencing Assay

A. Comparative expression of ASS1 in parental vs. parental and tamoxifen-resistant ILC cell lines as determined by change in Fragments Per Kilobase of transcript per Million mapped reads (FPKM) obtained from RNA seq data. B. qRT-PCR analysis showing ASS1 expression, and C. western blot and densitometry analysis showing ASS1 protein levels in MB134 and SUM44 cells, their respective tamoxifen resistant and LTED derivatives (LTEDA and LTEDB). D. Schematic diagram showing the CpG island in the ASS1 promoter. E. Analysis of the ASS1 promoter region using MS-PCR showing amplification of 150 bp methylated DNA in TAMR and LTED derivatives of ILC cell lines and 147 bp unmethylated DNA in parental ILC cell lines. F. qRT-PCR analysis showing ASS1 expression in MB134TAMR and SUM44TAMR cells that are either left untreated (UT) or treated with 5 μM 5-Aza-2’-deoxycytidine (dAzaC) for 120 hours. G. Western blot and densitometry analysis showing ASS1 protein levels in MB134TAMR and SUM44TAMR cells after 120 hours of dAzaC treatment (n=2 experiments). Error bars represent the standard deviation of triplicates. Significance levels are indicated as follows: ****p <0.0001, ***p <0.001, **p <0.01, *p <0.05.

Journal: bioRxiv

Article Title: Invasive lobular carcinoma integrated multi-omics analysis reveals silencing of Arginosuccinate synthase and upregulation of nucleotide biosynthesis in tamoxifen resistance

doi: 10.1101/2025.01.16.633236

Figure Lengend Snippet: A. Comparative expression of ASS1 in parental vs. parental and tamoxifen-resistant ILC cell lines as determined by change in Fragments Per Kilobase of transcript per Million mapped reads (FPKM) obtained from RNA seq data. B. qRT-PCR analysis showing ASS1 expression, and C. western blot and densitometry analysis showing ASS1 protein levels in MB134 and SUM44 cells, their respective tamoxifen resistant and LTED derivatives (LTEDA and LTEDB). D. Schematic diagram showing the CpG island in the ASS1 promoter. E. Analysis of the ASS1 promoter region using MS-PCR showing amplification of 150 bp methylated DNA in TAMR and LTED derivatives of ILC cell lines and 147 bp unmethylated DNA in parental ILC cell lines. F. qRT-PCR analysis showing ASS1 expression in MB134TAMR and SUM44TAMR cells that are either left untreated (UT) or treated with 5 μM 5-Aza-2’-deoxycytidine (dAzaC) for 120 hours. G. Western blot and densitometry analysis showing ASS1 protein levels in MB134TAMR and SUM44TAMR cells after 120 hours of dAzaC treatment (n=2 experiments). Error bars represent the standard deviation of triplicates. Significance levels are indicated as follows: ****p <0.0001, ***p <0.001, **p <0.01, *p <0.05.

Article Snippet: ILC cell lines MDA-MB-134-VI (MB134) (ATCC, USA) and SUM44PE (SUM44) (Asterand, USA) were used to develop the tamoxifen-resistant (TAMR) cells by continuously exposing them to 100 to 500 nM of 4-hydroxy tamoxifen (4-OHT) for over 6 months.

Techniques: Expressing, RNA Sequencing Assay, Quantitative RT-PCR, Western Blot, Amplification, Methylation, Standard Deviation

MB134 and SUM44 cells were transduced with plasmids expressing shRNA targeting ASS1 or the corresponding empty vector to produce shASS1 and control (pLKO) cell lines. A. qRT-PCR analysis showing ASS1 expression, and B. western blot and densitometry analysis of ASS1 protein levels in ASS1 knockdown ( shASS1 ) vs. pLKO (control) MB134 and SUM44 cell lines. C. Dose response to tamoxifen and IC 50 of tamoxifen in ASS1 knockdown ( shASS1 ) vs. pLKO (control) MB134 (left panel) and SUM44 cell lines (right panel). D . Comparison of growth kinetics of shASS1 vs. pLKO derivatives of MB134 (left panel) and SUM44 cell lines (right panel). Fold change in growth is normalized to day 0 at each time point over 72 hours. E. Overall survival analysis of all breast cancer patients in relation to ASS1 expression using METABRIC data set. F . Overall survival analysis of ILC patients who received endocrine therapy in relation to ASS1 expression using METABRIC data set. G. Overall survival analysis of breast cancer patients who received endocrine therapy in relation to ASS1 expression using K-M plotter dataset. ****p <0.0001, ***p <0.001, **p <0.01, *p <0.05.

Journal: bioRxiv

Article Title: Invasive lobular carcinoma integrated multi-omics analysis reveals silencing of Arginosuccinate synthase and upregulation of nucleotide biosynthesis in tamoxifen resistance

doi: 10.1101/2025.01.16.633236

Figure Lengend Snippet: MB134 and SUM44 cells were transduced with plasmids expressing shRNA targeting ASS1 or the corresponding empty vector to produce shASS1 and control (pLKO) cell lines. A. qRT-PCR analysis showing ASS1 expression, and B. western blot and densitometry analysis of ASS1 protein levels in ASS1 knockdown ( shASS1 ) vs. pLKO (control) MB134 and SUM44 cell lines. C. Dose response to tamoxifen and IC 50 of tamoxifen in ASS1 knockdown ( shASS1 ) vs. pLKO (control) MB134 (left panel) and SUM44 cell lines (right panel). D . Comparison of growth kinetics of shASS1 vs. pLKO derivatives of MB134 (left panel) and SUM44 cell lines (right panel). Fold change in growth is normalized to day 0 at each time point over 72 hours. E. Overall survival analysis of all breast cancer patients in relation to ASS1 expression using METABRIC data set. F . Overall survival analysis of ILC patients who received endocrine therapy in relation to ASS1 expression using METABRIC data set. G. Overall survival analysis of breast cancer patients who received endocrine therapy in relation to ASS1 expression using K-M plotter dataset. ****p <0.0001, ***p <0.001, **p <0.01, *p <0.05.

Article Snippet: ILC cell lines MDA-MB-134-VI (MB134) (ATCC, USA) and SUM44PE (SUM44) (Asterand, USA) were used to develop the tamoxifen-resistant (TAMR) cells by continuously exposing them to 100 to 500 nM of 4-hydroxy tamoxifen (4-OHT) for over 6 months.

Techniques: Transduction, Expressing, shRNA, Plasmid Preparation, Control, Quantitative RT-PCR, Western Blot, Knockdown, Comparison

A. Significantly enriched metabolic pathways in shASS1 vs. pLKO derivatives of MB134, and B. SUM44 cell lines. C. Changes in the levels of purine (dGMP, dGDP, GDP, GMP, IMP) and pyrimidine (UDP, UMP and CMP) nucleotides in parental (P) vs. tamoxifen-resistant (TAMR) MB134 cells. E. Changes in the levels of purine (ATP and ADP, dGTP, GDP) and F. pyrimidine (Cytosine, UMP) nucleotides in parental (P) vs. tamoxifen-resistant (TAMR) SUM44 cells. G. Expression levels of PRPS1 , H. PAICS, and I. DHODH in MB134 and SUM44 cell pairs as analyzed from RNA seq data. (FPKM: Fragments Per Kilobase Per Million reads). J. Representative picture of pCAD S1859 (pCAD) levels in parental and TAMR cells and K. in control (pLKO) vs. shASS1 derivatives of MB134 and SUM 44 cell pairs. Bar diagrams show average of more than one independent experiments (n=2). ***p <0.001, **p <0.01, *p <0.05.

Journal: bioRxiv

Article Title: Invasive lobular carcinoma integrated multi-omics analysis reveals silencing of Arginosuccinate synthase and upregulation of nucleotide biosynthesis in tamoxifen resistance

doi: 10.1101/2025.01.16.633236

Figure Lengend Snippet: A. Significantly enriched metabolic pathways in shASS1 vs. pLKO derivatives of MB134, and B. SUM44 cell lines. C. Changes in the levels of purine (dGMP, dGDP, GDP, GMP, IMP) and pyrimidine (UDP, UMP and CMP) nucleotides in parental (P) vs. tamoxifen-resistant (TAMR) MB134 cells. E. Changes in the levels of purine (ATP and ADP, dGTP, GDP) and F. pyrimidine (Cytosine, UMP) nucleotides in parental (P) vs. tamoxifen-resistant (TAMR) SUM44 cells. G. Expression levels of PRPS1 , H. PAICS, and I. DHODH in MB134 and SUM44 cell pairs as analyzed from RNA seq data. (FPKM: Fragments Per Kilobase Per Million reads). J. Representative picture of pCAD S1859 (pCAD) levels in parental and TAMR cells and K. in control (pLKO) vs. shASS1 derivatives of MB134 and SUM 44 cell pairs. Bar diagrams show average of more than one independent experiments (n=2). ***p <0.001, **p <0.01, *p <0.05.

Article Snippet: ILC cell lines MDA-MB-134-VI (MB134) (ATCC, USA) and SUM44PE (SUM44) (Asterand, USA) were used to develop the tamoxifen-resistant (TAMR) cells by continuously exposing them to 100 to 500 nM of 4-hydroxy tamoxifen (4-OHT) for over 6 months.

Techniques: Expressing, RNA Sequencing Assay, Control