Journal: PLoS ONE

Article Title: Photo-Activated Psoralen Binds the ErbB2 Catalytic Kinase Domain, Blocking ErbB2 Signaling and Triggering Tumor Cell Apoptosis

doi: 10.1371/journal.pone.0088983

Figure Lengend Snippet: (A) 8MOP interacts with three peptide regions within the ErbB2 catalytic kinase domain. Qualitative peptide identifications within the ErbB2 catalytic kinase domain following LC-MS/MS analysis of a streptavidin pull-down of biotinylated-8MOP bait (see ). The transmembrane domain is indicated (red diamond) and the five C-terminus tyrosine autophosphorylation sites are indicated (p). (B) Non-reducing Western blot analysis of the interaction of 8MOP with ErbB2. BT474 cells were treated with 800dye-8MOP (Promega) or with vehicle (0.01% DMSO) alone served as control for 48 hr and then exposed to UV irradiation (2J) prior to Western blot analysis. The image on the left shows the Western blot for ErbB2 (red). The image on the right shows the same membrane directly scanned for the presence of 800dye-8MOP (green), which overlays the ErbB2 signal. The results are representative of three independent experiments.

Article Snippet: ErbB2+ (BT474; SKBR3) and ErbB2 negative (MCF-7; T47D) human breast cancer cell lines, and the human foreskin fibroblast (HFF) cell line were obtained from the American Type Culture Collection (Manassas, VA).

Techniques: Liquid Chromatography with Mass Spectroscopy, Western Blot, Control, Irradiation, Membrane

Journal: PLoS ONE

Article Title: Photo-Activated Psoralen Binds the ErbB2 Catalytic Kinase Domain, Blocking ErbB2 Signaling and Triggering Tumor Cell Apoptosis

doi: 10.1371/journal.pone.0088983

Figure Lengend Snippet: The growth and viability of BT474 and SKBR3 cells (top bar graphs) after being subjected to the indicated treatment conditions. The combination of PUVA plus neratinib: P <0.0005 (BT474 and SKBR3 cells). Results represent the mean +/− standard error of triplicate samples, and are representative of three independent experiments. (B) Western blot analysis showing steady-state ErbB2, ErbB3, and phospho-Akt (S473) protein levels in BT474 and SKBR3 cells treated according to the indicated treatment conditions. Vehicle alone (0.01% DMSO) served as a control. Steady-state actin protein levels served as a control for equal loading of protein. The results are representative of three independent experiments.

Article Snippet: ErbB2+ (BT474; SKBR3) and ErbB2 negative (MCF-7; T47D) human breast cancer cell lines, and the human foreskin fibroblast (HFF) cell line were obtained from the American Type Culture Collection (Manassas, VA).

Techniques: Western Blot, Control

Journal: PLoS ONE

Article Title: Photo-Activated Psoralen Binds the ErbB2 Catalytic Kinase Domain, Blocking ErbB2 Signaling and Triggering Tumor Cell Apoptosis

doi: 10.1371/journal.pone.0088983

Figure Lengend Snippet: Top bar graph shows the results of the growth assays performed in T47D and stably transfected T47D cell line. T47D cells expressing p85 ErbB2 were pretreated with 5 µM lapatinib or 5 µM 8MOP for 4 hr followed by irradiation in a UV Stratalinker 1800 (Statagene). Cells transfected with empty vector (T47D/Vector), and those treated with vehicle alone (0.01% DMSO) served as controls. The effects of the treatments on cell growth and viability are shown in the bar graph. P<0.0071 (8MOP + UVA irradiation). Results represent the mean +/− standard error of triplicate samples, and are representative of three independent experiments. Steady-state phospho-p85 ErbB2 protein levels (dotted arrow) and phospho-p185 ErbB2 (solid arrow) are shown by Western blot. Actin steady-state protein levels served as a control to ensure for equal loading of protein. Results are representative of three independent experiments.

Article Snippet: ErbB2+ (BT474; SKBR3) and ErbB2 negative (MCF-7; T47D) human breast cancer cell lines, and the human foreskin fibroblast (HFF) cell line were obtained from the American Type Culture Collection (Manassas, VA).

Techniques: Stable Transfection, Transfection, Expressing, Irradiation, Plasmid Preparation, Western Blot, Control

Journal: Endocrinology

Article Title: ATM Is Required for the Prolactin-Induced HSP90-Mediated Increase in Cellular Viability and Clonogenic Growth After DNA Damage.

doi: 10.1210/en.2017-00652

Figure Lengend Snippet: Figure 3. D1-9-G129R-hPRL PRLR antagonist abrogates the PRL-mediated increase in SKBR3 cell viability. (A) Western blot of MCF7 and SKBR3 whole cell extracts probed with the anti-PRLR extracellular domain antibody, 1A2B1. (B) Western blot of nuclear extracts (10 mg/well) of SKBR3 cells starved in serum-free media for 16 hours then treated with 5 mg/mL oPRL or 5 mg/mL D1-9-G129R-hPRL PRLR antagonist (Antag) for 1 hour or left untreated, probed for p-STAT5, STAT5 (rabbit-anti-STAT5), and histone-H3 (HH3) (loading control). (C) A general schematic for treatments used in WST-1 assays including a 24-hour pretreatment, 2 hours of TOPOII poison, followed by a 48-hour recovery period. (D, E, and F) WST-1 assays using SKBR3 cells pretreated or not with 5 mg/mL oPRL 6 D1-9-G129R-hPRL for 24 hours followed by 2 hours of doxorubicin (DOX) treatment, with or without PRL or D1-9-G129R-hPRL, then a 48-hour recovery with or without PRL or D1-9-G129R-hPRL as indicated. (D) The effect of oPRL or PRLR antagonist alone. (E) The effect of DOX 6 PRL and (F) the effect of DOX + Antag 6 PRL. Graphs represents pooled experiments, n = 12. Data were analyzed with a one-way ANOVA followed by Bonferroni posttests. *P , 0.05; **P , 0.01; ***P , 0.001.

Article Snippet: Cell culture and cell lines PRL-responsive SKBR3 (HER2 overexpressing, p53mutant) and MCF7 (estrogen receptor+, p53 wild type) human breast cancer cell lines, obtained and authenticated from American Type Culture Collection, were used within 6 months when revived from frozen storage.

Techniques: Western Blot, Control

Journal: Endocrinology

Article Title: ATM Is Required for the Prolactin-Induced HSP90-Mediated Increase in Cellular Viability and Clonogenic Growth After DNA Damage.

doi: 10.1210/en.2017-00652

Figure Lengend Snippet: Figure 4. HSP90 inhibition abrogates PRL-mediated resistance. (A, B, and C) SKBR3 or (D, E, and F) MCF7 cells were pretreated or not with 5 mg/mL oPRL, 15 nM (SKBR3) or 100 nM (MCF7) HSP90 inhibitor 17-AAG, or a combination of the two for 24 hours followed by 2 hours of doxorubicin (DOX) treatment, 6PRL or 17-AAG, and a 48-hour recovery 6 PRL. (A–C) WST-1 assays of SKBR3 (A) with PRL or 17-AAG alone, (B) with DOX 6 PRL, or (C) with DOX + 17-AAG 6 PRL. (D–F) WST-1 assays of MCF7 (D) with PRL or 17-AAG alone, (E) with DOX 6 PRL, or (F) with DOX + 17-AAG 6 PRL. Graphs represent pooled experiments for SKBR3 n = 12, and for MCF7 n = 18. (G–I) WST-1 assays of MCF7 (G) with oPRL or 800 nM BIIB021 alone, (H) with DOX 6 PRL, or (I) with DOX + BIIB021 6 PRL. n = 18. Data were analyzed with a one-way ANOVA followed by Bonferroni posttests. *P , 0.05; **P , 0.01; ***P , 0.001.

Article Snippet: Cell culture and cell lines PRL-responsive SKBR3 (HER2 overexpressing, p53mutant) and MCF7 (estrogen receptor+, p53 wild type) human breast cancer cell lines, obtained and authenticated from American Type Culture Collection, were used within 6 months when revived from frozen storage.

Techniques: Inhibition

Journal: Endocrinology

Article Title: ATM Is Required for the Prolactin-Induced HSP90-Mediated Increase in Cellular Viability and Clonogenic Growth After DNA Damage.

doi: 10.1210/en.2017-00652

Figure Lengend Snippet: Figure 6. JAK2 inhibition abrogates the PRL increased cell viability. (A) Western blot of JAK2 inhibitor dose response. MCF7 cells were treated with twofold increasing concentrations of G6 for 12 hours. p-STAT5 levels were determined (mouse anti-p-STAT5) with total STAT5 and histone H3 (HH3) loading controls. Results are representative of two independent experimental replicates. (B) Dose-response curves of JAK2 inhibitor G6. MCF7 cells were treated with G6 at indicated time points and concentrations. The cell viability was determined with WST-1 cell viability assay. WST-1 assays of (C–E) MCF7 or (F–H) SKBR3 cells pretreated with 25 ng/mL hPRL for 24 hours or 25 mM JAK2 inhibitor G6 for 12 hours, followed by

Article Snippet: Cell culture and cell lines PRL-responsive SKBR3 (HER2 overexpressing, p53mutant) and MCF7 (estrogen receptor+, p53 wild type) human breast cancer cell lines, obtained and authenticated from American Type Culture Collection, were used within 6 months when revived from frozen storage.

Techniques: Inhibition, Western Blot, Viability Assay

Journal: Endocrinology

Article Title: ATM Is Required for the Prolactin-Induced HSP90-Mediated Increase in Cellular Viability and Clonogenic Growth After DNA Damage.

doi: 10.1210/en.2017-00652

Figure Lengend Snippet: Figure 7. HSP90 inhibition decreases ATM and p-ATM protein but not its mRNA. (A) MCF7 cells were treated or not with threefold increasing concentrations of 17-AAG for 24 hours followed by 2 hours of doxorubicin (DOX) treatment and recovery. Proteins (30 mg per well) were resolved on SDS-PAGE gel, and blot was probed for p-ATM, ATM, HSP90a, and GRB2. Gel is representative of two experiments. Fold difference of p-ATM with DOX treatment was calculated, in ImageJ, compared with untreated cells (set to 1). Fold difference of (B) total ATM and (C) p- ATM levels with 17-AAG and DOX treatment were calculated compared with DOX alone (set to 1). Representative blot of two experiments. (D) MCF7 cells were pretreated with oPRL (5 mg/mL) or 17-AAG for 24 hours, followed by 2 hours DOX treatment (0.2 mM). ATM expression was detected by qPCR, and the results were normalized to YWHAZ control [mean 6 standard error of the mean (SEM); n = 5]. The DDCt method was used to analyze the relative changes in gene. The letter “a” above SEM bars denotes that there were no statistically significant differences (one-way ANOVA followed by Bonferroni test, P , 0.05). (E) SKBR3 cells as above, with (F) ATM and (G) p-ATM band intensities quantified by ImageJ. Representative blot of two experiments.

Article Snippet: Cell culture and cell lines PRL-responsive SKBR3 (HER2 overexpressing, p53mutant) and MCF7 (estrogen receptor+, p53 wild type) human breast cancer cell lines, obtained and authenticated from American Type Culture Collection, were used within 6 months when revived from frozen storage.

Techniques: Inhibition, SDS Page, Expressing, Control

Journal: Endocrinology

Article Title: ATM Is Required for the Prolactin-Induced HSP90-Mediated Increase in Cellular Viability and Clonogenic Growth After DNA Damage.

doi: 10.1210/en.2017-00652

Figure Lengend Snippet: Figure 11. ATM is needed for PRL-induced increase in cell viability after DNA damage in 3D culture. Cells were pretreated with 25 ng/mL hPRL or vehicle for 24 hours followed by 2 hours of 1 mM doxorubicin (DOX) treatment (or vehicle) with or without PRL. Cells were transferred to collagen 3D culture and allowed to recover for 48 hours. Cell viability was determined with WST-1 assay, and the treatments were normalized to vehicle controls: (A) SKBR3 or (B) MCF7. For ATM knockdown experiments, (C and E) SKBR3 and (D and F) MCF7 cells were transfected with (C and D) siNT or (E and F) siATM, followed by 24 hours of 25 ng/mL hPRL and 2 hours of DOX treatment (3 mM for SKBR3, 2 mM for MCF7 cells). Cells were transferred to collagen gels to allow recovery for 96 hours (SKBR3) or 48 hours (MCF7). Bars in the graphs represent six independent experiments. Statistically significant analysis by ANOVA: *P , 0.05; **P , 0.01; ***P , 0.001.

Article Snippet: Cell culture and cell lines PRL-responsive SKBR3 (HER2 overexpressing, p53mutant) and MCF7 (estrogen receptor+, p53 wild type) human breast cancer cell lines, obtained and authenticated from American Type Culture Collection, were used within 6 months when revived from frozen storage.

Techniques: WST-1 Assay, Knockdown, Transfection

Journal: Journal of Cancer Research and Clinical Oncology

Article Title: New HER2-negative breast cancer subtype responsive to anti-HER2 therapy identified

doi: 10.1007/s00432-020-03144-7

Figure Lengend Snippet: Percent inhibition of NRG1b CELx signal for HER2+ cell lines versus HER2−/HSFs+ patient samples . Inhibition of NRG1-driven HER2 signaling by pertuzumab (10 mg/mL) or trastuzumab (10 mg/mL) alone or in combination in the CELx test. Percentages shown represent the average of four HER2+ HSFs+ cell lines (SKBR3, EFM192A, HCC1569, and ZR75-30) or 5 HER2−/HSFs+ primary tumor samples (C978, C133, C264, C309, and C371), performed with 2 technical replicates per biological sample

Article Snippet: Human breast cancer cell lines used in this study included SKBr3 (HER2+ /HSFs+), BT474 (HER2+ /HSFs−), BT483 (ER-/HER2−/HSFs+), ZR75-30 (HER2+ /HSFs+), HCC1569 (HER2+ /HSFs−), HCC1954 (ER-/HER2+ /HSFs−), HCC202 (HER2+ /HSFs+), MDA-MB361 (HER2+ /HSFs−), AU565 (HER2+ /HSFs−) (all from American Tissue Type Collection; ATCC, Manassas, VA), and EFM192A (HER2+ /HSFs+) (from Leibniz Institute DSMZ, Germany).

Techniques: Inhibition

Journal: Journal of Cancer Research and Clinical Oncology

Article Title: New HER2-negative breast cancer subtype responsive to anti-HER2 therapy identified

doi: 10.1007/s00432-020-03144-7

Figure Lengend Snippet: Comparison of inhibition of NRG1-driven HER2 signaling by HER2 targeted therapeutics. Pertuzumab (10 μg/mL), lapatinib (200 nM), afatinib (120 nM), or neratinib (500 nM) inhibition of NRG1-driven HER2 signaling is shown as the average of 9 HER2+ cell lines (SKBR3, EFM192A, ZR75-30, HCC202, HCC1954, HCC1569, MDA-MB361, BT474, AU565) or 7 HER2−/HER2s+ primary tumors (R69, R20, R160, R82, R95, R25, R71) performed with 2 technical replicates per biological sample. Single dose concentrations were selected for relative comparator purposes only where a maximal amount of the signaling was inhibited in at least one of the samples. Variances from 100% inhibition likely arise from ligand generated signaling that was not directly related to HER activity or where a sample had some level of drug paradoxical response (Claus et al. )

Article Snippet: Human breast cancer cell lines used in this study included SKBr3 (HER2+ /HSFs+), BT474 (HER2+ /HSFs−), BT483 (ER-/HER2−/HSFs+), ZR75-30 (HER2+ /HSFs+), HCC1569 (HER2+ /HSFs−), HCC1954 (ER-/HER2+ /HSFs−), HCC202 (HER2+ /HSFs+), MDA-MB361 (HER2+ /HSFs−), AU565 (HER2+ /HSFs−) (all from American Tissue Type Collection; ATCC, Manassas, VA), and EFM192A (HER2+ /HSFs+) (from Leibniz Institute DSMZ, Germany).

Techniques: Comparison, Inhibition, Generated, Activity Assay

Journal: American Journal of Translational Research

Article Title: Maytansine-loaded star-shaped folate-core PLA-TPGS nanoparticles enhancing anticancer activity

doi:

Figure Lengend Snippet: Fluorescence microscopy images images of SKBR3 after 4 h of incubation with the coumarin 6-loaded star FA-PLA-TPGS nanoparticles (coumarin 6-NPs). The coumarin 6-loaded nanoparticles were green (×100). A. SKBR3 cells incubated with coumarin 6-NPs for 12 hours; B. SKBR3 cells incubated with coumarin 6-NPs for 24 hours; C. SKBR3 cells incubated with coumarin 6-NPs for 36 hours.

Article Snippet: Human breast adenocarcinoma cell line SKBR3 (HER2-over expression) was obtained from American Type Culture Collection (ATCC; Rockville, MD, USA).

Techniques: Fluorescence, Microscopy, Incubation

Journal: American Journal of Translational Research

Article Title: Maytansine-loaded star-shaped folate-core PLA-TPGS nanoparticles enhancing anticancer activity

doi:

Figure Lengend Snippet: Growth inhibitory effects of maytansine and maytansine-NPs on SKBR3 cell lines in vitro. The effects of maytansine and maytansine-NPs (equivalent in maytansine) was tested at a concentrations 0.001, 0.01, 0.1, 1.0 and 10.0 nmol/L after treatment 48 h and 72 h, respectively. Both drug inhibited the growth of SKBR3 cells in a dose-dependent manner, but the effect on maytansine-NPs had no difference with that of maytansine in the concentration range of 0.01-10.0 nmol/L (p > 0.05).

Article Snippet: Human breast adenocarcinoma cell line SKBR3 (HER2-over expression) was obtained from American Type Culture Collection (ATCC; Rockville, MD, USA).

Techniques: In Vitro, Concentration Assay

Journal: Oncotarget

Article Title: MiR-1268b confers chemosensitivity in breast cancer by targeting ERBB2-mediated PI3K-AKT pathway

doi: 10.18632/oncotarget.20099

Figure Lengend Snippet: (A) Determined by RT-qPCR, miR-1268b was significantly upregulated in chemosensitive tissues by 9.6–fold compared with that of chemoresistant tissues ( *** p<0.0001). (B) miR-1268b was upregulated by 17.5–fold in ERBB2 negative tissues compared with that of ERBB2 positive tissues ( *** p<0.0001). (C) The expression of miR-1268b was detected in seven breast cancer cell lines. MiR-1268b was lower expressed in MCF-7/ADM (resistant to adrimycin), MDA-MB-453 and SKBR3 (Both are ERBB2 overexpression cells) in comparision with other cancer cells. In fact, the expression of miR-1268b in MCF-7/ADM was lower by 7.9-fold than that of MCF-7 cells ( *** p < 0.0001). (D, E) After transfected with miR-1268b mimics, expression of miR-1268b was significantly upregulated both in MDA-MB-231 and MDA-MB-468 cell lines, and transfected with miR-1268b inhibitor showed the opposite effect ( *** p < 0.0001).

Article Snippet: Human breast cancer cell lines MDA-MB-231, MDA-MB-468, MDA-MB-453 SKBR3 (MDA-MB-453 and SKBR3 are ERBB2 positive cell lines), MCF-7 were acquired from the American Type Culture Collection (ATCC) and maintained in Leibovitz's L15 medium supplemented with 10% fetal bovine serum (FBS, BI, Grand Island, NY, USA.).

Techniques: Quantitative RT-PCR, Expressing, Over Expression, Transfection

Journal: Oncotarget

Article Title: MiR-1268b confers chemosensitivity in breast cancer by targeting ERBB2-mediated PI3K-AKT pathway

doi: 10.18632/oncotarget.20099

Figure Lengend Snippet: (A) The software RNAhybrid predicted that ERBB2 was a potential target gene of miR-1268b, with the binding sites displayed. (B) The luciferase activity was reduced in pmirGLO-ERBB2 and miR-1268b co-transfected breast cancer cells in both MDA-MB 231 and MDA-MB-468, in comparison with that of the negative control ( ** p < 0.01, *** p < 0.001). (C, D, E, F) Western blot assay revealed that overexpression of miR-1268b could dramatically reduce the protein level of ERBB2 in MDA-MB-453 (C, D) and SKBR3 (E, F) ( * p < 0.05, ** p < 0.01). (G, H) The result of immunocytochemistry assay showed that miR-1268b transfection, rather than the negative control, could repress the protein expression of ERBB2 in MDA-MB-453 and SKBR3 cells.

Article Snippet: Human breast cancer cell lines MDA-MB-231, MDA-MB-468, MDA-MB-453 SKBR3 (MDA-MB-453 and SKBR3 are ERBB2 positive cell lines), MCF-7 were acquired from the American Type Culture Collection (ATCC) and maintained in Leibovitz's L15 medium supplemented with 10% fetal bovine serum (FBS, BI, Grand Island, NY, USA.).

Techniques: Software, Binding Assay, Luciferase, Activity Assay, Transfection, Comparison, Negative Control, Western Blot, Over Expression, Immunocytochemistry, Expressing

Journal: Oncotarget

Article Title: MiR-1268b confers chemosensitivity in breast cancer by targeting ERBB2-mediated PI3K-AKT pathway

doi: 10.18632/oncotarget.20099

Figure Lengend Snippet: (A, B) RT-qPCR results showed that miR-1268b could inhibit the expression of PIK3CA, AKT2, AKT3 and Bcl2 in MDA-MB-453 (A) and SKBR3 (B) ( * p <0.05, ** p <0.01, *** p<0.001). (C, D, E, F) Western blot assay further showed that miRNA-1268b reduced the protein expression of PIK3CA, AKT and Bcl2 in MDA-MB-453 (C, D) and SKBR3 (E, F) ( * p < 0.05). (G) The schematic diagram showed the potential functional mechanism of miR-1268b to induce chemosensitivity in breast cancer. MiR-1268b could repress the PI3K-AKT signaling pathway by targeting ERBB2 and inhibit the anti-apoptosis protein Bcl2.

Article Snippet: Human breast cancer cell lines MDA-MB-231, MDA-MB-468, MDA-MB-453 SKBR3 (MDA-MB-453 and SKBR3 are ERBB2 positive cell lines), MCF-7 were acquired from the American Type Culture Collection (ATCC) and maintained in Leibovitz's L15 medium supplemented with 10% fetal bovine serum (FBS, BI, Grand Island, NY, USA.).

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Functional Assay

Journal: Molecular Pharmaceutics

Article Title: Selective Cytotoxicity to HER2 Positive Breast Cancer Cells by Saporin-Loaded Nanobody-Targeted Polymeric Nanoparticles in Combination with Photochemical Internalization

doi: 10.1021/acs.molpharmaceut.8b01318

Figure Lengend Snippet: Cellular uptake of fluorescently labeled, targeted, and nontargeted placebo NPs. Confocal images of SkBr3 cells incubated with (A) Cys-NPs (22 μg/mL) and (B) 11A4-NPs (22 μg/mL) at different time points (nuclei are stained in blue, NPs are observed in orange, and the phase contrast image is in gray). The NPs remained in contact with the cells throughout the whole assay (no washing steps). (C) Mean fluorescence intensity in cytoplasm of SkBr3 and MDA-MB-231 cells after incubation with Cys-NPs and 11A4-NPs, data represent mean ± SD ( n = 15 imaging fields). Confocal images of SkBr3 cells (D) preincubated for 30 min with free 11A4 (12 μg/mL) followed by incubation for 30 min with 11A4-NPs (80 μg/mL) and (E) incubated for 30 min with 11A4-NPs alone (80 μg/mL). Nuclei are stained in blue, NPs are observed in orange, and the overlay of phase contrast image shows the cell borders. (F) Mean fluorescence intensity in cytoplasm of SkBr3 cells exposed to the competition conditions; data represent mean ± SD ( n = 15 imaging fields).

Article Snippet: Human breast cancer cell lines SkBr3 (HER2 positive, ATCC HTB-30) and MDA-MB-231 (HER2 negative, ATCC CRM-HTB-26) were obtained from American Type Culture Collection (Virginia, USA).

Techniques: Labeling, Incubation, Staining, Fluorescence, Imaging

Journal: Molecular Pharmaceutics

Article Title: Selective Cytotoxicity to HER2 Positive Breast Cancer Cells by Saporin-Loaded Nanobody-Targeted Polymeric Nanoparticles in Combination with Photochemical Internalization

doi: 10.1021/acs.molpharmaceut.8b01318

Figure Lengend Snippet: Cytotoxicity of TPPS 2a (0.5 μg/mL) combined with different illumination times. (A) Cell viability and (B) cell proliferation of SkBr3 and MDA-MB-231 cells exposed to PS and illumination, without NPs, relative to the nontreated control (no PS, no light). Data represent mean ± SD ( n = 6). (C) Cell viability and (D) cell proliferation of SkBr3 cells in the absence and presence of placebo 11A4-NPs and saporin-loaded 11A4-NPs relative to the nontreated control (no PS, no light, no NPs). Final NPs concentration in the wells was 90 μg/mL. Data represent mean ± SD ( n = 6 for tests in the absence of NPs, n = 3 tests in the presence of NPs). Illumination for 20, 40, and 60 s corresponds to light doses of ∼0.3, 0.5, and 0.8 J/cm 2 .

Article Snippet: Human breast cancer cell lines SkBr3 (HER2 positive, ATCC HTB-30) and MDA-MB-231 (HER2 negative, ATCC CRM-HTB-26) were obtained from American Type Culture Collection (Virginia, USA).

Techniques: Control, Concentration Assay

Journal: Molecular Pharmaceutics

Article Title: Selective Cytotoxicity to HER2 Positive Breast Cancer Cells by Saporin-Loaded Nanobody-Targeted Polymeric Nanoparticles in Combination with Photochemical Internalization

doi: 10.1021/acs.molpharmaceut.8b01318

Figure Lengend Snippet: Cytotoxicity of saporin-loaded NPs with or without PCI. The concentration of saporin reported in the graphs is 13% of the dose loaded in the NPs. Cytotoxicity of saporin-loaded NPs on (A,B) MDA-MB-231 and (C,D) SkBr3 cells. Cell viability and cell proliferation were calculated respective to the nontreated control (no PCI, no NPs). (E) Cell viability and (F) cell proliferation of SkBr3 cells incubated with free saporin and saporin-loaded NPs calculated respective to a control exposed to PCI in absence of protein and NPs. This control had 73 ± 5% cell viability and 73 ± 8% cell proliferation relative to a nontreated sample (no PCI, no free protein, no NPs). Data represent mean ± SD ( n = 3). Data Cys-NPs + PCI vs 11A4-NPs + PCI analyzed by unequal variances t test; * p -value < 0.05 and ** p -value < 0.001.

Article Snippet: Human breast cancer cell lines SkBr3 (HER2 positive, ATCC HTB-30) and MDA-MB-231 (HER2 negative, ATCC CRM-HTB-26) were obtained from American Type Culture Collection (Virginia, USA).

Techniques: Concentration Assay, Control, Incubation

Journal: Molecular Pharmaceutics

Article Title: Selective Cytotoxicity to HER2 Positive Breast Cancer Cells by Saporin-Loaded Nanobody-Targeted Polymeric Nanoparticles in Combination with Photochemical Internalization

doi: 10.1021/acs.molpharmaceut.8b01318

Figure Lengend Snippet: Cytotoxicity of saporin-loaded 11A4-NPs with PCI on SkBr3 cells with and without preincubation with free 11A4. (A) Cell viability and (B) cell proliferation were assessed in cells incubated with 11A4-NPs and subjected to PCI treatment or in cells preincubated with an excess of free 11A4 followed by the addition of 11A4-NPs and PCI treatment. Data represent mean ± SD ( n = 3). Data 11A4-NPs + PCI vs 11A4 + 11A4-NPs + PCI analyzed by unequal variances t test; * p -value < 0.05.

Article Snippet: Human breast cancer cell lines SkBr3 (HER2 positive, ATCC HTB-30) and MDA-MB-231 (HER2 negative, ATCC CRM-HTB-26) were obtained from American Type Culture Collection (Virginia, USA).

Techniques: Incubation

Journal: Molecular Pharmaceutics

Article Title: Selective Cytotoxicity to HER2 Positive Breast Cancer Cells by Saporin-Loaded Nanobody-Targeted Polymeric Nanoparticles in Combination with Photochemical Internalization

doi: 10.1021/acs.molpharmaceut.8b01318

Figure Lengend Snippet: Microscopic observations of SkBr3 cells at 40 and 120 h after treatment with saporin-loaded NPs and/or PCI. Representative images obtained by phase contrast and by overlay of the channels depicting the signals for Annexin V-FITC (apoptotic cells, in green) and PI (dead cells, in red). Scale bar = 100 μm.

Article Snippet: Human breast cancer cell lines SkBr3 (HER2 positive, ATCC HTB-30) and MDA-MB-231 (HER2 negative, ATCC CRM-HTB-26) were obtained from American Type Culture Collection (Virginia, USA).

Techniques: