Journal: bioRxiv

Article Title: Loss of Flotillin-2 enhances trastuzumab emtansine internalization and cytotoxicity by relieving negative regulation of HER2 internalization in HER2-amplified cancers

doi: 10.64898/2026.05.15.725439

Figure Lengend Snippet: A) HCC1954 cells were fixed, permeabilized, blocked, and probed with PLA probes against HER2 and FLOT2 combined, or either probe alone (negative controls) and DAPI (blue). Green signal indicates interaction between HER2 and FLOT2. Scale bar is 10 µm. B) SKBR3 cells were transfected with siNeg or siFLOT2 for seven days prior to cell lysing and immunoprecipitation with anti-FLOT2, followed by immunoblotting for HER2 or FLOT2. Whole cell lysate was immunoblotted for HER2, FLOT2 or HSC70 (loading control). C) HeLa control or FLOT2 KO cells were lysed and immunoprecipitated with anti-HER2, followed by immunoblotting for HER2 and FLOT2. Whole cell lysate was immunoblotted for HER2, FLOT2 and HSC70 (loading control). D) HEK293T cells were transfected with empty vector, HER2, FLAG-tagged FLOT2 or HER2 and FLAG-tagged FLOT2 for three days, then lysed and immunoprecipitated with anti-FLAG and immunoblotted for HER2 and FLAG. Whole cell lysate was immunoblotted for HER2, FLOT2 and HSC70 (loading control). E) SKBR3 cells were transfected with siNeg or siFLOT2 for five days prior to cell counting or three days prior to lysing and immunoblot to probe for pHER2 (Y1196), HER2, pMAPK (T202/Y204), ERK2, pAKT (S473), AKT, pS6 (S235/236), S6, FLOT2, and HSC70 (loading control). Graphed data represent the average ±SEM of at least three independent experiments, and statistical analysis was performed by Student’s t-test. F) Same as in E, with HCC202 cells transfected for seven days prior to cell counting or three days prior to lysing, and lysates were instead probed for pHER2 (Y1221/1222). G) Same as in E, with HCC1954 cells transfected for seven days prior to cell counting and three days prior to lysing.

Article Snippet: The HER2 amplified breast cancer cell lines SKBR3, HCC1954, BT474, HCC1419, HCC2218, UACC893 were obtained from ATCC.

Techniques: Transfection, Immunoprecipitation, Western Blot, Control, Plasmid Preparation, Cell Counting

Journal: bioRxiv

Article Title: Loss of Flotillin-2 enhances trastuzumab emtansine internalization and cytotoxicity by relieving negative regulation of HER2 internalization in HER2-amplified cancers

doi: 10.64898/2026.05.15.725439

Figure Lengend Snippet: A) SKBR3 shFLOT2 cells were treated +/- 500 ng/mL doxycycline for 72 hours, replated, then treated with pHrodo-T-DM1 (1 µg/mL; red) in serum-free RPMI for 7 hours. Cells were stained with CellTracker Blue CMAC dye (1 µM; Blue) in serum-free RPMI for 30 minutes prior to confocal imaging. GFP (green) expression is induced by doxycycline, indicating induction of the shFLOT2 promoter. Intensity of pHrodo signal per cell was quantified and divided by the area of each cell (right). Data represents the average ±SEM of at least three independent experiments, and statistical analysis was performed by Student’s t-test. Scale bar is 40 µm. B) Same as in A, with SKBR3 shControl cells. GFP (green) expression is induced by doxycycline, indicating induction of the shControl promoter. Scale bar is 20 µm. C) SKBR3 cells were transfected with siNeg or siFLOT2 for 48 hours and then treated with DMSO or 1 ng/mL T-DM1 for 24 hours in 1% FBS RPMI. Cells were lysed and immunoblotted for HER2, pAKT (S473), AKT, pS6 (S235/236), S6, FLOT2 and HSC70 (loading control). Band density was normalized to loading control, and then normalized to siNeg DMSO for each individual protein. Data represents the average ±SEM of at least three independent experiments, and statistical analysis was performed by Student’s t-test. D) Same as in C, with HCC202 cells using 10 µg/mL T-DM1.

Article Snippet: The HER2 amplified breast cancer cell lines SKBR3, HCC1954, BT474, HCC1419, HCC2218, UACC893 were obtained from ATCC.

Techniques: Staining, Imaging, Expressing, Transfection, Control

Journal: bioRxiv

Article Title: Loss of Flotillin-2 enhances trastuzumab emtansine internalization and cytotoxicity by relieving negative regulation of HER2 internalization in HER2-amplified cancers

doi: 10.64898/2026.05.15.725439

Figure Lengend Snippet: A) SKBR3 shFLOT2 or shControl were pretreated with DMSO control (black) or 500 ng/mL doxycycline (gray) for 48 hours in 1% FBS RPMI, then treated with indicated T-DM1 concentration with continued doxycycline in 1% FBS RPMI for three days prior to cell counting. The table below the graphs indicates the calculated IC50 comparing DMSO + T-DM1 to doxycycline + T-DM1. Data represents the average ±SEM of at least three independent experiments, and statistical analysis was performed by Student’s t-test. B) Same as in A, with HCC202 shFLOT2 or shControl. C) Same as in A, with HCC1954 shFLOT2 or shControl. D) ARK1 (top) or ARK2 (bottom) cells were transfected with siNeg (black) or siFLOT2 (gray) for 48 hours prior to being treated with indicated T-DM1 concentration in 10% FBS RPMI for three days prior to cell viability reading (CellTiter-Glo 2.0). Data represents the average ±SEM of at least three independent experiments, and statistical analysis was performed by Student’s t-test. E) SKBR3, HCC202, HCC1954, BT474, HCC1419, HCC2218, UACC893, ARK1, and ARK2 cells were treated with 0-1000 ng/mL T-DM1 in 1% FBS RPMI for three days prior to cell counting. Average FLOT2 expression by immunoblot (relative to HSC70 loading control) and the calculated IC50 for each cell line was plotted. Protein expression and IC50 were calculated from at least three independent experiments. Line represents linear regression. F) Same as E, with average HER2 expression (relative to HSC70 loading control). G) Overall survival of patients in Caris dataset from T-DXd to last contact with high FLOT2 (orange; 16.22 months) vs low FLOT2 (blue; 18.26 months), p=0.041. H) Overall survival of patients in Caris dataset from T-DM1 to last contact with high FLOT2 (orange; 37.967 months) vs low FLOT2 (blue; 41.29 months), p=0.131.

Article Snippet: The HER2 amplified breast cancer cell lines SKBR3, HCC1954, BT474, HCC1419, HCC2218, UACC893 were obtained from ATCC.

Techniques: Control, Concentration Assay, Cell Counting, Transfection, Expressing, Western Blot

Journal: bioRxiv

Article Title: Loss of Flotillin-2 enhances trastuzumab emtansine internalization and cytotoxicity by relieving negative regulation of HER2 internalization in HER2-amplified cancers

doi: 10.64898/2026.05.15.725439

Figure Lengend Snippet: A) SKBR3 shFLOT2 cells were treated +/- 500 ng/mL doxycycline for a total of four days, and transfected with HA-tagged Ubiquitin for a total of three days. Cells were then treated with 10 ng/mL T-DM1 in 1% FBS RPMI for 2 hours, lysed, immunoprecipitated with anti-HA antibody, and immunoblotted for HER2 and HA. Whole cell lysate was immunoblotted for HER2, HA, FLOT2 and HSC70 (loading control). B) SKBR3 (left) or HCC1954 (right) were treated with TAK-243 (1 nM for SKBR3, 100 nM for HCC1954) and T-DM1 (10 ng/mL for SKBR3, 100 ng/mL for HCC1954) in 1% FBS RPMI for three days. Dead cell percentage was calculated using PI stain with the BioTek Cytation. Data represents the average ±SEM of at least three independent experiments, and statistical analysis was performed by Student’s t-test. C) SKBR3 cells were treated with DMSO (control), 1 nM or 10 nM TAK-243 for 24 hours in 1% FBS RPMI, and then treated with pHrodo-T-DM1 (1 µg/mL; red) for 7 hours in serum-free RPMI. Cells were stained with CellTracker Blue CMAC dye (1 µM; Blue) for 30 minutes in serum-free RPMI prior to confocal imaging. Intensity of pHrodo signal per cell was quantified and divided by the area of each cell (right). Data represents the average ±SEM of at least three independent experiments, and statistical analysis was performed by Student’s t-test. Scale bar is 40 µm. D) SKBR3 shFLOT2 were pretreated +/- 500 ng/mL doxycycline in complete RPMI for 48 hours, and then continued doxycycline +/- TAK-243 (1 nM) for 24 hours in 1% FBS RPMI. Cells were then treated with pHrodo-T-DM1 in serum-free RPMI (1 µg/mL; red) for 7 hours. Cells were stained with CellTracker Blue CMAC dye (1 µM; Blue) in serum-free RPMI for 30 minutes prior to confocal imaging. GFP (green) expression is induced by doxycycline, indicating induction of the shFLOT2 promoter. Intensity of pHrodo signal per cell was quantified and divided by the area of each cell (right). Data represents the average ±SEM of at least three independent experiments, and statistical analysis was performed by Student’s t-test. Scale bar is 20 µm.

Article Snippet: The HER2 amplified breast cancer cell lines SKBR3, HCC1954, BT474, HCC1419, HCC2218, UACC893 were obtained from ATCC.

Techniques: Transfection, Ubiquitin Proteomics, Immunoprecipitation, Control, Staining, Imaging, Expressing

Journal: bioRxiv

Article Title: Loss of Flotillin-2 enhances trastuzumab emtansine internalization and cytotoxicity by relieving negative regulation of HER2 internalization in HER2-amplified cancers

doi: 10.64898/2026.05.15.725439

Figure Lengend Snippet: A) SKBR3 shFLOT2 cells were transfected with siRNA for Cbl (siCbl) or neg control (siNeg)and +/- 500 ng/mL doxycycline for 4 days total and transfected with HA-tagged ubiquitin for 3 days total. Cells were then treated with 10 ng/mL for 2 hours in 1% FBS RPMI, lysed, immunoprecipitated with anti-HA antibody, and immunoblotted for HER2, HA, and ubiquitin. Whole cell lysate was immunoblotted for HA, Cbl, FLOT2 and HSC70 (loading control). Immunoprecipitation band intensity was quantified and HER2 and ubiquitin and graphed as HER2/ubiquitin (right). siNeg with T-DM1 and doxycycline was normalized to siNeg with DMSO, and siCbl with T-DM1 and doxycycline was normalized to siCbl with DMSO. Data represents the average ±SEM of at least three independent experiments, and statistical analysis was performed by Student’s t-test. B) SKBR3 shFLOT2 cells were transfected with siNeg or siCbl +/- 500 ng/mL doxycycline for 48 hours and re-plated on chambered coverglass, then treated with pHrodo-T-DM1 (1 µg/mL; red) in serum-free RPMI for 7 hours. Cells were stained with CellTracker Blue CMAC dye (1 µM; Blue) in serum-free RPMI for 30 minutes prior to confocal imaging. GFP (green) expression is induced by doxycycline, indicating induction of the shFLOT2 promoter. Intensity of pHrodo signal per cell was quantified and divided by the area of each cell (right). Data represents the average ±SEM of at least three independent experiments, and statistical analysis was performed by one-way ANOVA. Scale bar is 20 µm. C) HCC202, HCC1954 and SKBR3 lysates were immunoblotted for Cbl, Cbl-b and HSC70 (loading control). D) The same SKBR3 shFLOT2 cells that were transfected with siNeg or siCbl and treated +/-doxycycline for 48 hours in B were then re-plated and treated +/- 500 ng/mL doxycycline +/- 100 ng/mL T-DM1 for an additional 72 hours and dead cell percentage was calculated using PI stain with the BioTek Cytation. Cells were lysed and immunoblotted (bottom) for Cbl, FLOT2 and HSC70 (loading control) to confirm knockdown. Data represents the average ±SEM of at least three independent experiments, and statistical analysis was performed by Student’s t-test. Dead cell percentage was normalized to the baseline percentage recorded at time of TDM1 treatment. E) Same as in D, with HCC1954 shFLOT2 cells. SiCbl was co-transfected with si-Cbl-b and cells were treated with 1000 ng/mL T-DM1, and lysates were also immunoblotted for cbl-b.

Article Snippet: The HER2 amplified breast cancer cell lines SKBR3, HCC1954, BT474, HCC1419, HCC2218, UACC893 were obtained from ATCC.

Techniques: Transfection, Control, Ubiquitin Proteomics, Immunoprecipitation, Staining, Imaging, Expressing, Knockdown

Journal: bioRxiv

Article Title: Loss of Flotillin-2 enhances trastuzumab emtansine internalization and cytotoxicity by relieving negative regulation of HER2 internalization in HER2-amplified cancers

doi: 10.64898/2026.05.15.725439

Figure Lengend Snippet: A) SKBR3 cells were lysed, lysates treated with DMSO or 250 µM zoledronic acid (ZA) for an hour, and then heated at the indicated temperatures for 3 minutes each. Lysates were then centrifuged, and supernatant was immunoblotted for FLOT2, PHB1, and PHB2 (left). Quantification of bands and calculation of melting temperatures are depicted on the right. Quantification of bands represent the average ±SEM of three independent experiments. B) SKBR3 cells were treated with 0, 1 or 10 µM zoledronic acid for 24 or 48 hours, lysed, immunoprecipitated by anti-HER2, and immunoblotted for HER2, FLOT2, PHB1 or PHB2. Whole cell lysate was immunoblotted for HER2, FLOT2, PHB1, PHB2 and HSC70 (loading control). Quantification of immunoprecipitation bands from three independent experiments for FLOT2 relative to HER2 are depicted on the right, with the average ±SEM. Statistical analysis was performed by Student’s t-test compared to 0 µM zoledronic acid. C) SKBR3 cells were treated with DMSO (control) or 1 µM Zoledronic acid (ZA) for 72 hours then treated with pHrodo-T-DM1 (1 µg/mL; red) in serum-free RPMI for 7 hours. Cells were stained with CellTracker Blue CMAC dye (1 µM; Blue) in serum-free RPMI for 30 minutes prior to confocal imaging. Intensity of pHrodo signal per cell was quantified and divided by the area of each cell (right). Data represents the average ±SEM of at least three independent experiments, and statistical analysis was performed by Student’s t-test. Scale bar is 20 µm. D) SKBR3 cells were treated with the indicated combinations of DMSO (control), 1 ng/mL T-DM1 and 1 µM zoledronic acid (ZA) for 48 hours in 1% FBS RPMI. Dead cell percentage was calculated using PI stain with the BioTek Cytation. Data represents the average ±SEM of three independent experiments, and statistical analysis was performed by Student’s t-test. E) Same as D, with ARK1 cells in 10% FBS RPMI. F) Same as E, with ARK2 cells and 10 µM zoledronic acid (ZA). G) Same as D, with HCC1954 cells with 24 hour treatment.

Article Snippet: The HER2 amplified breast cancer cell lines SKBR3, HCC1954, BT474, HCC1419, HCC2218, UACC893 were obtained from ATCC.

Techniques: Immunoprecipitation, Control, Staining, Imaging

Journal: International Journal of Molecular Medicine

Article Title: Ebastine targets HER2/HER3 signaling and cancer stem cell traits to overcome trastuzumab resistance in HER2-positive breast cancer

doi: 10.3892/ijmm.2026.5751

Figure Lengend Snippet: EBA impairs cancer stem cell-like properties. (A) BT474 and SKBR3 cells were treated with EBA for 48 h, and ALDH1 activity was assessed by flow cytometry using the Aldefluor assay. DEAB was used to define the baseline of Aldefluor-positive fluorescence. (B) BT474 cells (5x10 4 cells/ml) were plated in ultra-low attachment dishes and cultured in the presence or absence of EBA for 5 days. The number and volume of mammospheres were measured by microscopy. (C) Overall survival of patients with breast cancer stratified by the co-expression of ALDH1A1 and CD44. (D) Spearman correlation analysis of ALDH1A1 and CD44 mRNA levels in patients with HER2-positive breast cancer from The Cancer Genome Atlas cohort (n=76). Kaplan-Meier survival analyses of patients with HER2-overexpressing breast cancer stratified by (E) ALDH1A1 and (F) CD44 expression. Patients were divided into high- and low-expression groups based on the median gene expression. Statistical significance was determined using the log-rank test. (G) JIMT-1 cells were treated with EBA (3 μ M) for 48 h and the CD44 high /CD24 low cell populations were identified by flow cytometry. (H) JIMT-1 cells (1.5x10 4 cells/ml) were cultured under serum-free suspension conditions in the presence of EBA (3 μ M) for 8 days. Mammosphere number and volumes were quantified. ** P<0.01 and **** P<0.0001 vs. vehicle-treated control (0 μ M EBA). EBA, ebastine; ALDH, aldehyde dehydrogenase; DEAB, diethylaminobenzaldehyde; CTL, control; ISO, isotype.

Article Snippet: The human breast cancer cell lines SKBR3, BT474, MDA-MB-453 (American Type Culture Collection) and JIMT-1 (Leibnitz Institute DSMZ-German Collection of Microorganisms and Cell Cultures GmbH) were cultured in DMEM, MEM or RPMI-1640 (all Sigma-Aldrich; Merck KGaA) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) and 100 U/ml penicillin-streptomycin at 37°C in a humidified atmosphere of 5% CO 2 .

Techniques: Activity Assay, Flow Cytometry, Fluorescence, Cell Culture, Microscopy, Expressing, Gene Expression, Suspension, Control

Journal: International Journal of Molecular Medicine

Article Title: Ebastine targets HER2/HER3 signaling and cancer stem cell traits to overcome trastuzumab resistance in HER2-positive breast cancer

doi: 10.3892/ijmm.2026.5751

Figure Lengend Snippet: EBA impairs cancer stem cell-like properties. (A) BT474 and SKBR3 cells were treated with EBA for 48 h, and ALDH1 activity was assessed by flow cytometry using the Aldefluor assay. DEAB was used to define the baseline of Aldefluor-positive fluorescence. (B) BT474 cells (5x10 4 cells/ml) were plated in ultra-low attachment dishes and cultured in the presence or absence of EBA for 5 days. The number and volume of mammospheres were measured by microscopy. (C) Overall survival of patients with breast cancer stratified by the co-expression of ALDH1A1 and CD44. (D) Spearman correlation analysis of ALDH1A1 and CD44 mRNA levels in patients with HER2-positive breast cancer from The Cancer Genome Atlas cohort (n=76). Kaplan-Meier survival analyses of patients with HER2-overexpressing breast cancer stratified by (E) ALDH1A1 and (F) CD44 expression. Patients were divided into high- and low-expression groups based on the median gene expression. Statistical significance was determined using the log-rank test. (G) JIMT-1 cells were treated with EBA (3 μ M) for 48 h and the CD44 high /CD24 low cell populations were identified by flow cytometry. (H) JIMT-1 cells (1.5x10 4 cells/ml) were cultured under serum-free suspension conditions in the presence of EBA (3 μ M) for 8 days. Mammosphere number and volumes were quantified. ** P<0.01 and **** P<0.0001 vs. vehicle-treated control (0 μ M EBA). EBA, ebastine; ALDH, aldehyde dehydrogenase; DEAB, diethylaminobenzaldehyde; CTL, control; ISO, isotype.

Article Snippet: The human breast cancer cell lines SKBR3, BT474, MDA-MB-453 (American Type Culture Collection) and JIMT-1 (Leibnitz Institute DSMZ-German Collection of Microorganisms and Cell Cultures GmbH) were cultured in DMEM, MEM or RPMI-1640 (all Sigma-Aldrich; Merck KGaA) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) and 100 U/ml penicillin-streptomycin at 37°C in a humidified atmosphere of 5% CO 2 .

Techniques: Activity Assay, Flow Cytometry, Fluorescence, Cell Culture, Microscopy, Expressing, Gene Expression, Suspension, Control

Journal: BMC Cancer

Article Title: GCNT3 and ST3GAL1 expression correlates with HER2 status and MUC1/β-catenin/Cyclin D1 axis in breast cancer

doi: 10.1186/s12885-026-15821-w

Figure Lengend Snippet: In silico analysis showing differential expression of glycogenes in the progression of BC: A Gene Set Enrichment Analysis (GSEA) showing the heatmap for different glycogenes in the HER2 high vs low phenotype. B Pie chart made using PANTHER software representing: cellular pathways of genes overexpressed in glycosylation geneset, and biological pathways of genes overexpressed in glycosylation geneset, C Cellular pathways of genes overexpressed in KRAS geneset and biological pathways of genes overexpressed in KRAS geneset. Major pathways highlighted in these pie charts have been indicated

Article Snippet: HER2 positive cell line SKBR3 (ATCC Catalog HTB-30) was cultured in McCoy’s 5 A medium.

Techniques: In Silico, Quantitative Proteomics, Software, Glycoproteomics

Journal: BMC Cancer

Article Title: GCNT3 and ST3GAL1 expression correlates with HER2 status and MUC1/β-catenin/Cyclin D1 axis in breast cancer

doi: 10.1186/s12885-026-15821-w

Figure Lengend Snippet: In vitro analysis of ST3GAL1’s association with HER2: A Bar plot showing qRT-PCR analysis of ST3GAL1 in HER2 -ve and HER2 +ve BC cells. T-test based p-value obtained were as follows: SKBR3 vs MCF7 (p=0.0045, n=3), SKBR3 vs MDAMB231 (p=0.0034, n=3) and SKBR3 vs MDAMB435 (p=0.05, n=3). B Representative images of immunohistochemical analysis for ST3GAL1 in Grade 2 and 3 of Stage I, II and III BC tissues ( C ) ST3GAL1 expression in HER2 positive BC tissue. D Mean intensity value calculated as average pixel intensity using ImageJ, for ST3GAL1. Two-way ANOVA p-value exhibited p<0.05 (n=3) for Grade 2, Stage I vs Stage II vs Stage III. Similarly, p <0.05 (n=3) was obtained for Grade 3, Stage I vs Stage II vs Stage III. Moreover, t-test based p<0.05 (n=3) was obtained in the HER2 positive and HER2 negative BC tissues

Article Snippet: HER2 positive cell line SKBR3 (ATCC Catalog HTB-30) was cultured in McCoy’s 5 A medium.

Techniques: In Vitro, Quantitative RT-PCR, Immunohistochemical staining, Expressing

Journal: BMC Cancer

Article Title: GCNT3 and ST3GAL1 expression correlates with HER2 status and MUC1/β-catenin/Cyclin D1 axis in breast cancer

doi: 10.1186/s12885-026-15821-w

Figure Lengend Snippet: In silico and in vitro analyses showing inverse association of GCNT3 with HER2: A Survival plot representing the gene expression of GCNT3 in BC cohorts. B Differential expression analysis of GCNT3 using the RNAseq data for the tumor vs control BC. C Differential expression analysis for the control vs tumor samples from the TCGA BC dataset for GCNT3 using GEPIA. D Bar plot showing qRT-PCR analysis for GCNT3 using the HER2 -ve and HER2 +ve BC cells with T-test based p-value obtained as follows: SKBR3 vs MCF7 (p=0.045, n=3), SKBR3 vs MDAMB231 (p=0.012, n=3) and SKBR3 vs MDAMB435 (p=0.00069, n=3). E Representative images of immunohistochemical analysis for GCNT3 in Grade 2 and 3 of Stage I, II and III BC tissues. F , G Mean intensity values calculated as average pixel intensity using ImageJ, for GCNT3. Two-way ANOVA p-value exhibited p<0.05 for Grade 2, Stage I vs Stage II vs Stage III. Similarly, p<0.05 (n=3) was obtained for Grade 3, Stage I vs Stage II vs Stage III. Additionally, t-test based p<0.05 (n=3) was obtained in the HER2 negative vs positive BC tissues

Article Snippet: HER2 positive cell line SKBR3 (ATCC Catalog HTB-30) was cultured in McCoy’s 5 A medium.

Techniques: In Silico, In Vitro, Gene Expression, Quantitative Proteomics, RNA sequencing, Control, Quantitative RT-PCR, Immunohistochemical staining

Journal: BMC Cancer

Article Title: GCNT3 and ST3GAL1 expression correlates with HER2 status and MUC1/β-catenin/Cyclin D1 axis in breast cancer

doi: 10.1186/s12885-026-15821-w

Figure Lengend Snippet: In silico analysis of ST3GAL1’s association with HER2: A Survival plots representing gene expression of HER2 and ST3GAL1 for these genes in BC cohorts. B Oncomine extracted data represented by box plots on ST3GAL1 with increasing IHC score ( C ) Violin plots representing differential expression analysis of HER2 and ST3GAL for the tumor vs control using the RNAseq data for the BC samples. D Correlation plot between ST3GAL1 and HER2 on R2 dataset. E Differential expression analysis for the control vs tumor samples from the TCGA BC dataset for HER2 and ST3GAL1 using GEPIA. F Correlation plot between HER2 and ST3GAL1 on the cbioportal database

Article Snippet: HER2 positive cell line SKBR3 (ATCC Catalog HTB-30) was cultured in McCoy’s 5 A medium.

Techniques: In Silico, Gene Expression, Quantitative Proteomics, Control, RNA sequencing

Journal: BMC Cancer

Article Title: GCNT3 and ST3GAL1 expression correlates with HER2 status and MUC1/β-catenin/Cyclin D1 axis in breast cancer

doi: 10.1186/s12885-026-15821-w

Figure Lengend Snippet: Functional analysis using GCNT3 inhibitor (talniflumate) in HER2 -ve and HER2 +ve BC cells: Representative images and quantification using talniflumate for ( A ) wound healing on MCF7 cells, t-test based p-value for control vs treatment at 24h and 48h were p=0.38 (n=3) and p=0.17 respectively, B colony formation on MCF7 cells, C wound healing on MDAMB231 cells, t-test based p-value for control vs treatment at 24h and 48h were p=0.023 (n=3) and p=0.084 (n=3) respectively, D colony formation on MDAMB231 cells, E wound healing on MDAMB435 cells, t-test based p-value for control vs treatment at 24h and 48h were p=0.09 (n=3) and p=0.40 (n=3) respectively, F colony formation on MDAMB435 cells, G . wound healing for SKBR3 cells, t-test based p-value for control vs treatment at 12h and 24h were p=0.49 (n=3) and p=0.32 (n=3) respectively and H colony formation for SKBR3 cells. NOTE: T-test based p-value calculation for colony formation assay were found to be p<0.05 (n=3) for all the cell lines

Article Snippet: HER2 positive cell line SKBR3 (ATCC Catalog HTB-30) was cultured in McCoy’s 5 A medium.

Techniques: Functional Assay, Control, Colony Assay

Journal: BMC Cancer

Article Title: GCNT3 and ST3GAL1 expression correlates with HER2 status and MUC1/β-catenin/Cyclin D1 axis in breast cancer

doi: 10.1186/s12885-026-15821-w

Figure Lengend Snippet: Clinical significance of MUC1, β-catenin, Cyclin D1 in BC specimens: Representative images of immunohistochemical analysis in different stages and grades of BC for ( A ) β-catenin ( B ) MUC1 and C Cyclin D1. Mean intensity calculated of ( D ) β-catenin analyzed using two-way ANOVA p-value exhibited p<0.05 (n=3) for Grade 2, Stage I vs Stage II vs Stage III and Grade 3, Stage I vs Stage II vs Stage III, E MUC1 analyzed usingtwo-way ANOVA p-value exhibited p<0.05 (n=3) for Grade 2, Stage I vs Stage II, III and p>0.05 (n=3) for Grade 3, Stage I vs Stage II vs Stage III and F Cyclin D1 analyzed using two-way ANOVA p-value exhibited p<0.05 (n=3) for Grade 2, Stage I vs Stage II vs Stage III; however p>0.05 (n=3) was found for Grade 3 tissues. All the BC tissues herein were HER2 -ve

Article Snippet: HER2 positive cell line SKBR3 (ATCC Catalog HTB-30) was cultured in McCoy’s 5 A medium.

Techniques: Immunohistochemical staining

Journal: BMC Cancer

Article Title: GCNT3 and ST3GAL1 expression correlates with HER2 status and MUC1/β-catenin/Cyclin D1 axis in breast cancer

doi: 10.1186/s12885-026-15821-w

Figure Lengend Snippet: Multiple analyses of GCNT3 in driving the MUC1/β-catenin/Cyclin D1 axis in BC: Representative immunohistochemistry images and mean intensity plots of HER2 +ve and HER2 -ve BC samples for (A ) β-catenin ( B ) MUC1 and C Cyclin D1. D Correlation plots of MUC1 with GCNT3, β-catenin, Cyclin D1 and ST3GAL1. E Gene interacting plot for GCNT3 with top 20 interacting nodes using STRING. F Scheme of the mechanistic link between GCNT3 and the β-catenin/Cyclin D1 axis in breast cancer. NOTE: T-test based p-value was found to be significant only for the Cyclin D1 stained HER2 positive vs negative BC tissues (p<0.05, n=3) whereas they were insignificant for the β-catenin and MUC1

Article Snippet: HER2 positive cell line SKBR3 (ATCC Catalog HTB-30) was cultured in McCoy’s 5 A medium.

Techniques: Immunohistochemistry, Staining