Journal: Advanced Science

Article Title: Mitigating Doxorubicin‐Induced Cardiotoxicity and Enhancing Anti‐Tumor Efficacy with a Metformin‐Integrated Self‐Assembled Nanomedicine

doi: 10.1002/advs.202415227

Figure Lengend Snippet: Design and mechanism of PMDDH. A) Schematic representation of metformin enhancing the cytotoxic effect of Dox on 4T1 cells and mitigating its cardiotoxicity on H9c2 cells. B) Synthesis of PMD, PMDD, and PMDDH. C) Illustration of the targeted delivery, enhanced therapeutic efficacy, and reduced cardiotoxicity of PMDDH in vivo.

Article Snippet: Meanwhile, the 4T1 cells were collected and resuspended in a Cell Staining Buffer (Elabscience, Wuhan, China).

Techniques: Drug discovery, In Vivo

Journal: Advanced Science

Article Title: Mitigating Doxorubicin‐Induced Cardiotoxicity and Enhancing Anti‐Tumor Efficacy with a Metformin‐Integrated Self‐Assembled Nanomedicine

doi: 10.1002/advs.202415227

Figure Lengend Snippet: A) Visualization of cellular uptake of Dox, free FAM‐DNA, and PMDDH by CLSM. Scale bar = 20 µm. B,C) Flow cytometry analysis and corresponding quantitative results of PMDDH uptake in 4T1 cells ( n = 3). D) Visualization of CD44 expression (green) and PMDDH uptake (red) in 4T1, MDA‐MB‐231, and MCF‐7 cells. Scale bar = 20 µm. E) Dose‐dependent cytotoxicity of Dox, PMDH, and PMDDH in 4T1 cells ( n = 5). F) IC 50 values of Dox and PMDDH in 4T1 cells ( n = 5). G) Crystal violet staining of treated 4T1 cells showing cell density and distribution changes. H) CRT expression (red) in 4T1 cells post‐treatment. Scale bar = 50 µm. I) Flow cytometry analysis of CRT expression and J) corresponding quantitative results ( n = 3). K) HMGB1 levels in the supernatant of 4T1 cells following treatment ( n = 3). L) ATP release from 4T1 cells after 24 h of different treatments ( n = 3). Data are presented as mean ± SD (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: Meanwhile, the 4T1 cells were collected and resuspended in a Cell Staining Buffer (Elabscience, Wuhan, China).

Techniques: Flow Cytometry, Expressing, Staining

Journal: Advanced Science

Article Title: Mitigating Doxorubicin‐Induced Cardiotoxicity and Enhancing Anti‐Tumor Efficacy with a Metformin‐Integrated Self‐Assembled Nanomedicine

doi: 10.1002/advs.202415227

Figure Lengend Snippet: A) Schematic illustration of cGAS‐STING and AMPK signaling activation in 4T1 cells. B) Western blot analysis of p‐IRF3 and p‐STING levels in 4T1 cells after different treatments. RT‐qPCR analysis of cytokine and chemokine expression: C) IFN‐β, D) CXCL10, E) OAS1, F) ISG15, and G) TNF‐α in 4T1 cells. H) Western blot results showing p‐AMPK and p‐mTOR levels in 4T1 cells post‐treatment. I) Flow cytometry analysis and J) immunofluorescence imaging of PD‐L1 expression in 4T1 cells following various treatments. Scale bar = 20 µm. Data are presented as mean ± SD ( n = 3, * p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Meanwhile, the 4T1 cells were collected and resuspended in a Cell Staining Buffer (Elabscience, Wuhan, China).

Techniques: Activation Assay, Western Blot, Quantitative RT-PCR, Expressing, Flow Cytometry, Immunofluorescence, Imaging

Journal: Advanced Science

Article Title: Mitigating Doxorubicin‐Induced Cardiotoxicity and Enhancing Anti‐Tumor Efficacy with a Metformin‐Integrated Self‐Assembled Nanomedicine

doi: 10.1002/advs.202415227

Figure Lengend Snippet: A) Pharmacokinetics of free Dox and PMDDH in rats, with inset showing the drug‐time curve of the distribution phase ( n = 3). B) In vivo distribution of free Cy5‐DNA compared to Cy5‐labeled PMDDH. C) Ex vivo fluorescence imaging of tumors and major organs in tumor‐bearing mice following intravenous administration of free Cy5‐DNA and PMDDH. D) Schematic of treatment regimen for 4T1 tumor‐bearing mice. E) Tumor growth curves after various treatments over 16 days ( n = 5). F) Tumor inhibitory rates and G) tumor weights at day 16 ( n = 5). H) Histological evaluation of tumor tissues using H&E staining, Ki67 immunohistochemistry, and TUNEL immunofluorescence. Scale bars = 100 µm for H&E staining; 200 µm for Ki67 immunohistochemistry and TUNEL immunofluorescence. Data are presented as mean ± SD (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Meanwhile, the 4T1 cells were collected and resuspended in a Cell Staining Buffer (Elabscience, Wuhan, China).

Techniques: Drug discovery, In Vivo, Labeling, Ex Vivo, Fluorescence, Imaging, Staining, Immunohistochemistry, TUNEL Assay, Immunofluorescence

Journal: Scientific Reports

Article Title: Luciferase Expression Allows Bioluminescence Imaging But Imposes Limitations on the Orthotopic Mouse (4T1) Model of Breast Cancer

doi: 10.1038/s41598-017-07851-z

Figure Lengend Snippet: Growth rates of the murine adenocarcinoma 4T1 cell line and its sublclones expressing luciferase 4T1luc2D6 and 4T1luc2. The time course of the proliferation of the parental 4T1, and 4T1luc2D6 and 4T1luc2 (PerkinElmer) clones as determined using Nikon Biostation CT (Nikon, Japan) with STDV ( A ); Bioluminescence from 4T1luc2 and 4T1luc2D6 cells in culture assessed by bioluminescent imaging (Spectrum CT, Perkin Elmer) ( B ); Average level of bioluminescence of 4T1luc2 and 4T1luc2D6 cells (photons/cell/sec) ( C ). Results from five independent measurements. No significant difference between any of the analysed parameters (p > 0,05; Mann Whitney test).

Article Snippet: 4T1 (CRL-2539TM, ATCC) and 4T1luc2 (BW124087, Perkin Elmer, Waltham, MA) were purchased; 4T1luc-D6 clone was described previously .

Techniques: Expressing, Luciferase, Clone Assay, Imaging, MANN-WHITNEY

Journal: Scientific Reports

Article Title: Luciferase Expression Allows Bioluminescence Imaging But Imposes Limitations on the Orthotopic Mouse (4T1) Model of Breast Cancer

doi: 10.1038/s41598-017-07851-z

Figure Lengend Snippet: Growth in BALB/c mice (n = 5–6 per group) of primary tumors induced by the implantation of 4T1, 4T1luc2 and 4T1lucD6 cells (see Methods for description). Growth curves obtained by MRI visualize average tumor volume in cubic mm, with STDV ( A ); Growth of primary tumors induced by implantation of 4T1luc2 and 4T1lucD6 cells monitored by bioluminescent imaging; curves depict an average photon flux from the tumor per sec, with STDV ( B ); Statistical comparison of the tumor sizes evaluated by MRI ( C ). Median size of 4T1lucD6 tumors is significantly lower (p = 0,017), and of 4T1luc2 tumors tend to be lower than of 4T1 tumors, although the difference is not significant (p = 0,17). Day of implantation is counted as day 0. Statistical comparisons are done using Kruskal Wallis and Mann-Whitney tests (Statistica AXA 10.0).

Article Snippet: 4T1 (CRL-2539TM, ATCC) and 4T1luc2 (BW124087, Perkin Elmer, Waltham, MA) were purchased; 4T1luc-D6 clone was described previously .

Techniques: Imaging, Comparison, MANN-WHITNEY

Journal: Scientific Reports

Article Title: Luciferase Expression Allows Bioluminescence Imaging But Imposes Limitations on the Orthotopic Mouse (4T1) Model of Breast Cancer

doi: 10.1038/s41598-017-07851-z

Figure Lengend Snippet: Comparison of the luciferase activity measured as the intensity of luminescence (in arbitrary units a.u.; Enspire, Perkin Elmer) in the original 4T1luc2 and 4T1luc2D6 cell lines and in cell cultures prepared from the 4T1luc2 and 4T1luc2D6 tumors by the experimental end-point. Luciferase activity in the 4T1luc2 and 4T1luc2D6 cells prior to implantation ( A ); Luciferase activity in primary cell cultures prepared from tumors formed in BALB/c mice by implantation of 4T1luc2D6 (n = 3; 6 tumors) ( B ) or 4T1luc2 cells (n = 3; 6 tumors) ( C ); Recalculation of the average luciferase activity per cell with STDV ( D ). Individual tumors are coded by the cell line, mouse and tumor numbers, for example “4T1luc2D6 n10 r1” designate a tumor caused by implantation of cell line 4T1luc2D6 into mouse No. 10 and refer to sample No. 1 from one of the tumors. Curves in panels B and D show dependence of luciferase activity on the number of cells used in the assay; data represent the average of three parallel measurements. Statistical comparisons are done using Mann-Whitney test (Statistica AXA 10.0); **p < 0,01, and *p < 0,05.

Article Snippet: 4T1 (CRL-2539TM, ATCC) and 4T1luc2 (BW124087, Perkin Elmer, Waltham, MA) were purchased; 4T1luc-D6 clone was described previously .

Techniques: Comparison, Luciferase, Activity Assay, MANN-WHITNEY

Journal: Scientific Reports

Article Title: Luciferase Expression Allows Bioluminescence Imaging But Imposes Limitations on the Orthotopic Mouse (4T1) Model of Breast Cancer

doi: 10.1038/s41598-017-07851-z

Figure Lengend Snippet: In vitro IFN-gamma response of lymphocytes of BALB/c mice implanted with Luc-expressing 4T1luc2D6 or 4Tl1luc2, or parental 4T1 cells, to stimulation with peptide GFQSMYTFV representing an immunodominant CTL epitope of luciferase (LucP) assessed using IFN-γ ELISpot (Mabtech). IFN-γ response to LucP by the experimental end-point assessed as the average number of IFN-γ spot forming cells per mln splenocytes (SFC/mln) with STDV; *p < 0.05, IFN-γ response to LucP in BALB/c mice implanted with 4T1luc2D6 and 4T1luc2 cells compared to IFN-γ response exhibited by naïve or 4T1-implanted mice (Mann-Whitney test) ( A ); Development of cellular immune response to LucP in mice implanted with 5000 4T1luc2 cells, on days 6, 9 and 23 post implantation; *p < 0.05, IFN-γ response to LucP in BALB/c mice implanted with 4T1luc2 cells on days 9 and 23 post implantation compared to IFN-γ response exhibited by naïve mice or mice implanted by 4T1luc2 and assessed on day 6 post implantation (Friedman ANOVA and Kendall Coeff. of Concordance) ( B ). Red line indicates the cut-off for the specific IFN-γ response, defined as an average number of lymphocytes producing IFN-γ in response to stimulation with LucP ± 3 STDV per mln lymphocytes. Cut-off was established in independent in vitro tests done on splenocytes of naïve mice (n = 5).

Article Snippet: 4T1 (CRL-2539TM, ATCC) and 4T1luc2 (BW124087, Perkin Elmer, Waltham, MA) were purchased; 4T1luc-D6 clone was described previously .

Techniques: In Vitro, Expressing, Luciferase, Enzyme-linked Immunospot, MANN-WHITNEY

Journal: Scientific Reports

Article Title: Luciferase Expression Allows Bioluminescence Imaging But Imposes Limitations on the Orthotopic Mouse (4T1) Model of Breast Cancer

doi: 10.1038/s41598-017-07851-z

Figure Lengend Snippet: Intradermal Luc DNA immunization followed by electroporation (EP) with driving pulses of 100 V makes BALB/c mice resistant to initiation of 4T1luc2 tumors. BALB/c mice were immunized by plasmid encoding firefly luciferase pVaxLuc delivered by intradermal injections followed by EP using multineedle (Mn) electrodes mounted on Dermavax (Cellectis) (n = 4) or CUY21EditII (BEX Ltd) electroporators at 100 V (n = 6) or 50 V (n = 4). Immunization (red circles) and 4T1luc2 implantation sites (blue circles) visualized by in vivo imaging (Spectrum CT) on days 1 and 6 post tumor cell implantation (43 and 48 days of the immunization cycle). Mice immunized with Luc DNA (A, upper panel) versus empty vector (A, lower panel). Text boxes demonstrate total flux from the respective regions of interest (ROI) (photons/sec) ( A ); Quantification of bioluminescence emission (BLI; photon flux/sec) from immunization sites illustrates the efficacy of Luc gene transfer and expression. From day 5, Luc DNA immunized mice electroporated at 100 V emitted stronger bioluminescence than mice electroporated at 50 V (p < 0,05; Mann Whitney test), no bioluminescence from immunization sites in control mice receiving pVax1 ( B ); Quantification of bioluminescence (photon flux/sec) from 4T1luc2 implantation sites in mice immunized with Luc DNA and electroporated with DermaVax/Mn at 100 V (n = 4), BEX/Mn at 100 V (n = 6), BEX/Mn at 50 V (n = 4), and vector immunized mice (n = 3) on days 0 to 6 post challenge ( C ); Difference (in %) in BLI from 4T1luc2 implantation sites after six days of tumor growth in mice immunized with Luc DNA with EP at 100 V (DermaVax/Mn + BEX/Mn groups, 10 animals, dubbed “Luc, 100 V”), or 50 V (BEX/Mn, 4 animals, dubbed “Luc, 50 V”), vector immunized (n = 3) and naive mice (n = 5, from an independent experiment) ( D ). *p < 0,05; **p < 0,1 (Statistica AXA 10.0).

Article Snippet: 4T1 (CRL-2539TM, ATCC) and 4T1luc2 (BW124087, Perkin Elmer, Waltham, MA) were purchased; 4T1luc-D6 clone was described previously .

Techniques: Electroporation, Plasmid Preparation, Luciferase, In Vivo Imaging, Expressing, MANN-WHITNEY, Control

Journal: Scientific Reports

Article Title: Luciferase Expression Allows Bioluminescence Imaging But Imposes Limitations on the Orthotopic Mouse (4T1) Model of Breast Cancer

doi: 10.1038/s41598-017-07851-z

Figure Lengend Snippet: Cellular immune response to peptide GFQSMYTFV representing an immunodominant CTL epitope of luciferase (LucP; ref. ) in mice immunized with Luc DNA (pVaxLuc). The average number of IFN-γ producing splenocytes per mln cells in mice receiving pVaxLuc followed by electroporation using Dermavax (n = 6), or BEX devices at 100 V (n = 6), or empty vector pVax1 with electroporation using BEX device at 100 V (n = 6) after priming (day 21), boosting (day 30), and after challenge with 4T1luc2 cells (day 9, or 43 from the start) (see Methods for details) ( A ); FACS of the pooled splenocytes of Luc-gene immunized mice detecting LucP-specific secretion of IFN-γ by CD8 + T cells ( B ); Average LucP-specific IFN-γ response of mice immunized with Luc DNA and electroporated using CUY21EditII (BEX) at 50 V and 100 V ( C ). Red line indicates the cut-off for specific IFN-γ response as number of spots registered in non-immune (control and vector immunized) mice + 3 STDV. *p < 0,05, Luc DNA immunized mice electroporated at 100 V, and **p < 0,1, Luc DNA immunized mice electroporated at 50 V compared to vector immunized mice (Mann-Whitney test; STATISTICA AXA 10.0).

Article Snippet: 4T1 (CRL-2539TM, ATCC) and 4T1luc2 (BW124087, Perkin Elmer, Waltham, MA) were purchased; 4T1luc-D6 clone was described previously .

Techniques: Luciferase, Electroporation, Plasmid Preparation, Control, MANN-WHITNEY

Journal: Journal of Cellular and Molecular Medicine

Article Title: Obesity and triple‐negative‐breast‐cancer: Is apelin a new key target?

doi: 10.1111/jcmm.15639

Figure Lengend Snippet: Infusion of obesity‐related levels of apelin favours TNBC brain metastatization. A, Ex vivo bioluminescent signals from 4T1 Luc‐GFP in lungs of PBS or apelin‐infused (0.1 µmol/kg/d) Balb/c nude mice. B, Quantification of bioluminescent signals of lung metastases by intensity. C, Ex vivo bioluminescent signals from 4T1 Luc‐GFP in brains of PBS or Apelin‐infused Balb/c nude mice. D, Quantification of metastatic brains in PBS or Apelin‐infused mice. Number of mice per group for (A, B): Control: 9, Apelin: 9, and for (C, D): Control: 9, Apelin: 10. Data were analysed using two‐way ANOVA followed by Bonferroni post hoc test for (B). Data were analysed using Fisher's exact test for (D)

Article Snippet: The 4T1 Luc‐GFP cell line was obtained from Perkin Elmer (4T1 Red FLuc‐GFP, PerkinElmer, Waltham, MA, USA) and grows only in immunodeficient mice.

Techniques: Ex Vivo

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Combining photodynamic therapy and cascade chemotherapy for enhanced tumor cytotoxicity: the role of CTT 2 P@B nanoparticles

doi: 10.3389/fbioe.2024.1361966

Figure Lengend Snippet: ROS generation detection in vitro . (A) The release of singlet oxygen from each drug was detected in vitro under dark conditions. Data were presented as the mean ± SD ( n = 3). (B) Detection of singlet oxygen release of each drug under 660 nm laser irradiation at a power density of 280 mW cm -2 was performed in vitro . Data were presented as the mean ± SD ( n = 3). (C) ROS generation was observed in 4T1 cells treated with each medication while being exposed to laser irradiation at a wavelength of 660 nm and intensity of 280 mW cm −2 . Bright: Bright field. DCF: Green fluorescence represents intracellular ROS. Merge: Superimpose the image. Scale Bar = 100 μm.

Article Snippet: The murine breast cancer cells (4T1) were procured from Icell Biotech Co., Ltd. (Shanghai, China).

Techniques: In Vitro, Irradiation, Fluorescence

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Combining photodynamic therapy and cascade chemotherapy for enhanced tumor cytotoxicity: the role of CTT 2 P@B nanoparticles

doi: 10.3389/fbioe.2024.1361966

Figure Lengend Snippet: Evaluation of mitochondrial targeting function. (A) Confocal images of the mitochondrial sites of TPPOH 2 , CTT 2 , CTT 2 P, and CTT 2 P@B NPs in 4T1 cells. Mito-Tracker Green was used to stain the mitochondria in the green channel. The red channel was derived from the emission of the photosensitizer fraction (PS) itself. Merge stands for superimposed image. The Green and red curves in the Plot Profile represent the gray value of Mito-Tracker Green and PS, respectively. Scale bar = 20 μm. (B) Flow cytometry JC-1 method was used to analyze the mitochondrial function of cells treated with different drugs. Red fluorescence: normal mitochondria (J-aggregate); Green fluorescence: depolarized mitochondria (J-monomer).

Article Snippet: The murine breast cancer cells (4T1) were procured from Icell Biotech Co., Ltd. (Shanghai, China).

Techniques: Staining, Derivative Assay, Flow Cytometry, Fluorescence

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Combining photodynamic therapy and cascade chemotherapy for enhanced tumor cytotoxicity: the role of CTT 2 P@B nanoparticles

doi: 10.3389/fbioe.2024.1361966

Figure Lengend Snippet: Evaluation of cell death in vitro through cytotoxicity assessment and examination of apoptosis and necrosis. (A) The in vitro cytotoxicity of 4T1 cells treated with CPT, TPPOH 2 , CTT 2 , CTT 2 P, and CTT 2 P@B NPs in the dark was assessed using the CCK-8 assay. Data were presented as the mean ± SD ( n = 5). ** p < 0.01, **** p < 0.0001. (B) The in vitro cytotoxicity of 4T1 cells treated with TPPOH 2 , CTT 2 , CTT 2 P and CTT 2 P@B NPs under laser irradiation (660 nm, 280 mW cm −2 ) was determined using the CCK-8 assay. Data were presented as the mean ± SD ( n = 5). ** p < 0.01, **** p < 0.0001. (C) Cell apoptosis and necrosis were analyzed using flow cytometry with Annexin V-FITC/PI double staining following treatment with various drugs at a concentration of 5 μM.

Article Snippet: The murine breast cancer cells (4T1) were procured from Icell Biotech Co., Ltd. (Shanghai, China).

Techniques: In Vitro, CCK-8 Assay, Irradiation, Flow Cytometry, Double Staining, Concentration Assay

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Combining photodynamic therapy and cascade chemotherapy for enhanced tumor cytotoxicity: the role of CTT 2 P@B nanoparticles

doi: 10.3389/fbioe.2024.1361966

Figure Lengend Snippet: Biodistribution in vivo . (A) Blood compatibility test of CPT, TPPOH 2 , CTT 2 , CTT 2 P, CTT 2 P@B NPs. Data were presented as the mean ± SD ( n = 3). *** p < 0.001, **** p < 0.0001. (B) Time-lapse live fluorescence imaging of mice with 4T1 tumors following the administration of free CTT 2 P and CTT 2 P@B NPs via intravenous injection. (C) Fluorescent images of major organs and tumors were obtained 24 h after injection, using ex vivo methods. (D) The mean fluorescence intensity of each organ and tumor was used to determine the biodistribution of free CTT 2 P and CTT 2 P@B NPs in mice. Data were presented as the mean ± SD ( n = 3). * p < 0.05.

Article Snippet: The murine breast cancer cells (4T1) were procured from Icell Biotech Co., Ltd. (Shanghai, China).

Techniques: In Vivo, Fluorescence, Imaging, Injection, Ex Vivo

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Combining photodynamic therapy and cascade chemotherapy for enhanced tumor cytotoxicity: the role of CTT 2 P@B nanoparticles

doi: 10.3389/fbioe.2024.1361966

Figure Lengend Snippet: In vivo anti-tumor study of each drug in 4T1 tumor-bearing mice. (A) Tumor images of different drug administration treatments after the antitumor study. (B) During the administration, the growth of tumors in mice was observed in each group receiving treatment. Data were presented as the mean ± SD ( n = 5), ** p < 0.01. (C) The weight of mice in each treatment group was monitored throughout the administration period. Data were presented as the mean ± SD ( n = 5).

Article Snippet: The murine breast cancer cells (4T1) were procured from Icell Biotech Co., Ltd. (Shanghai, China).

Techniques: In Vivo

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Combining photodynamic therapy and cascade chemotherapy for enhanced tumor cytotoxicity: the role of CTT 2 P@B nanoparticles

doi: 10.3389/fbioe.2024.1361966

Figure Lengend Snippet: Investigation of the effects of each medication on mice with 4T1 tumors through pathological examination. (A) After administering various medications, the major organs (including the heart, liver, spleen, lung, and kidney) were subjected to H&E staining. Scale bar = 50 μm. (B) After administering various medications, tumors were subjected to H&E staining and TUNEL staining. Scale bar = 50 μm.

Article Snippet: The murine breast cancer cells (4T1) were procured from Icell Biotech Co., Ltd. (Shanghai, China).

Techniques: Medications, Staining, TUNEL Assay

Journal: Cancer research

Article Title: HDAC6 plays a non-canonical role in the regulation of anti-tumor immune responses, dissemination, and invasiveness of breast cancer.

doi: 10.1158/0008-5472.CAN-19-3738

Figure Lengend Snippet: (A) Growth kinetics of 4T1 implanted subcutaneously to the flank of female BALB/c mice, treated with different doses of NextA (1.0 to 25mg/kg), 5X/week. (B) Growth kinetics of 4T1 implanted orthotopically in 4th mammary gland, treated with different doses of NextA, 5X/week. (C) Quantification of lung nodules in vector control vs. 25mg/kg NextA treated mice, 5X/week. (D) Pictorial representation of lung nodules. (E) Tumor burden measured by Luciferin K-salt injected bioluminescent mice. (F) Quantification of surface nodules as counted per cubic mm of tumor volume. (G) Growth kinetics of non-target vs. HDAC6KD-4T1 tumors implanted orthotopically into mice. (H) Quantification of lung nodules from non-target vs. HDAC6KD-4T1 tumor-bearing host. (I) Pictorial representation of lung nodules.

Article Snippet: 4T1 cell line, a model for triple-negative murine carcinoma, was a generous gift from Dr. Scott Abrams’ laboratory at RPCI, Buffalo, NY (authenticated by IDEXX through STR-profiling: details provided as supplemental document ).

Techniques: Plasmid Preparation, Control, Injection

Journal: Cancer research

Article Title: HDAC6 plays a non-canonical role in the regulation of anti-tumor immune responses, dissemination, and invasiveness of breast cancer.

doi: 10.1158/0008-5472.CAN-19-3738

Figure Lengend Snippet: (A) Gene expression of EMT-molecules with or without in vitro NextA treatment (5μM) on 4T1 cells by qPCR. (B) Expression of EMT-genes with or without in vitro NextA (5μM) on metastatic 4T1 lung-isolated clones by qPCR. (C) Expression of EMT-genes after in vitro NextA treatment (5μM) on MDA-MB-231 cells by qPCR. (D) Expression of EMT-genes after in vitro NextA treatment (5μM) on MCF7 cells by qPCR. (E) Evaluation of E-Cad levels in 4T1 cells after in vitro NextA treatment (5μM) by western blot. Evaluation of acetylated tubulin as an internal indicator for NextA mediated acetylation function. (F) Detection of the endogenous levels of E-Cad by western blot from in vivo tumor samples treated with NextA (25mg/kg). (G) Endogenous expression of HDAC6 and E-Cad protein in non-target vs. HDAC6KD-4T1 clones. (H) Corresponding E-Cad RNA levels in non-target vs. HDAC6KD-4T1 clones.. (I) Expression of EMT-genes in non-target and HDAC6KD-4T1 cells. (J) NextA dose-dependent invasion by 4T1 cells measured in 2D invasion chamber. GM-6001, a broad-spectrum MMP-inhibitor, is used as a positive control. (K) Quantification of wound closure of 4T1 cells throughout 16h, with or without NextA, as analyzed by ImageJ. (L) Snapshots of wound closure of 4T1 cells throughout 16h, with or without NextA, captured on Leica DMi8 microscope with a piezo stage (Pecon) with phase-contrast.

Article Snippet: 4T1 cell line, a model for triple-negative murine carcinoma, was a generous gift from Dr. Scott Abrams’ laboratory at RPCI, Buffalo, NY (authenticated by IDEXX through STR-profiling: details provided as supplemental document ).

Techniques: Gene Expression, In Vitro, Expressing, Isolation, Clone Assay, Western Blot, In Vivo, Positive Control, Microscopy

Journal: Cancer research

Article Title: HDAC6 plays a non-canonical role in the regulation of anti-tumor immune responses, dissemination, and invasiveness of breast cancer.

doi: 10.1158/0008-5472.CAN-19-3738

Figure Lengend Snippet: (A) Growth kinetics of 4T1 tumors implanted in the 4th mammary gland of female BALB/c mice, treated with different doses of αPD-1 antibody (0.5 to 15mg/kg). (B) Quantification of lung nodules in vehicle control vs. αPD-1 treated mice. (C) Gene expression of IFNγ and PD-L1 in the in vivo tumors collected from the mice with or without αPD-1 treatment measured by qPCR. (D) Expression of PD-L1 gene in 4T1 cells treated in vitro with 5μM of NextA measured by qPCR. (E) Expression of PD-L1 on tumor cells co-cultured with CD3+ splenocytes from non-tumor bearing mice and αPD-1 antibody, with or without treatment with NextA or IFNγ-neutralizing antibody, measured by qPCR. (F) Expression of PD-L1 on tumor cells co-cultured with CD3+ splenocytes from 4T1 tumor-bearing mice and αPD-1 antibody, with or without treatment with NextA or IFNγ-neutralizing antibody, measured by qPCR.

Article Snippet: 4T1 cell line, a model for triple-negative murine carcinoma, was a generous gift from Dr. Scott Abrams’ laboratory at RPCI, Buffalo, NY (authenticated by IDEXX through STR-profiling: details provided as supplemental document ).

Techniques: Control, Gene Expression, In Vivo, Expressing, In Vitro, Cell Culture

Journal: Cancer research

Article Title: HDAC6 plays a non-canonical role in the regulation of anti-tumor immune responses, dissemination, and invasiveness of breast cancer.

doi: 10.1158/0008-5472.CAN-19-3738

Figure Lengend Snippet: (A) Schematic representation of combination treatment. (B) Growth kinetics of 4T1 tumors implanted in the 4th mammary gland of female BALB/c mice, treated with the predetermined doses of NextA and αPD-1; 6X/week. (C) Growth kinetics of the individual tumors in each treatment group. (D) Quantification of lung nodules from each treatment group. (E) Pictorial representation of lung nodules from each group.

Article Snippet: 4T1 cell line, a model for triple-negative murine carcinoma, was a generous gift from Dr. Scott Abrams’ laboratory at RPCI, Buffalo, NY (authenticated by IDEXX through STR-profiling: details provided as supplemental document ).

Techniques:

Journal: Cancer research

Article Title: HDAC6 plays a non-canonical role in the regulation of anti-tumor immune responses, dissemination, and invasiveness of breast cancer.

doi: 10.1158/0008-5472.CAN-19-3738

Figure Lengend Snippet: (A) Measurement of E-Cad gene expression by qPCR in 4T1 cells in vitro treated with E-Cad siRNA. (B) Measurement of E-Cad protein expression by Western blot in 4T1 cells in vitro treated with E-Cad siRNA. (C) Measurement of E-Cad protein expression by Western blot in 4T1 cells in vitro treated with E-Cad SiRNA and NextA (5μM). (D) Invasion capability of E-Cad silenced 4T1 cells at different concentrations of NextA (2.5 and 5μM) measured in 2D invasion chamber. (E) Expression of ZEB1 in the 4T1 cells after NextA (5μM) and Stattic (10μM) treatment, measured by qPCR. (F) MDA-MB-231 cells were treated with NextA (5μM), and the expression of E-Cad, ZEB1, and tubulin, evaluated by immunoblot. (G) A regulatory model is proposing HDAC6is as modulators of the STAT3/ZEB1 axis and subsequent regulation of the expression of E-Cadherin.

Article Snippet: 4T1 cell line, a model for triple-negative murine carcinoma, was a generous gift from Dr. Scott Abrams’ laboratory at RPCI, Buffalo, NY (authenticated by IDEXX through STR-profiling: details provided as supplemental document ).

Techniques: Gene Expression, In Vitro, Expressing, Western Blot

Journal: Cells

Article Title: Interaction between Chromodomain Y-like Protein and Androgen Receptor Signaling in Sertoli Cells Accounts for Spermatogenesis

doi: 10.3390/cells13100851

Figure Lengend Snippet: AR is colocalized and interacts with CDYL in male testes and Sertoli cells. ( A ) Immunohistochemical staining of AR and CDYL in testes obtained from wild-type and ARKO mice. Bar = 50 um. ( B ) Localization of AR and CDYL (upper) and co-localization of AR, CDYL, and DAPI (bottom) in the testes obtained from wild-type mice by immunofluorescence analysis. Bar = 20 μm. ( C ) Interaction was observed between AR and CDYL in wild-type mouse testes. IgG was used as the control for Western blotting in each group. “Input” means the sample on 10% of volume used for IP. ( D ) Protein expression patterns of AR and CDYL in wild-type mouse testes and ARKO mice by Western blotting. GAPDH was used as the internal control. ( E ) AR and CDYL mRNA expressions were detected in testicular tissues between wild-type and ARKO mice by quantitative RT–PCR assay. ( n ≥ 3) * p ˂ 0.05, by unpaired two-tailed Student t tests was significant compared with the control. Data are expressed as the mean ± standard error of three samples per group. S: Sertoli cell; L: Leydig cell; M: myoid cell; SP: spermatids.

Article Snippet: The TM4 mouse Sertoli cell line (BCRC-60254) was procured from the Bioresource Collection and Research Center (BCRC) in Hsinchu, Taiwan.

Techniques: Immunohistochemical staining, Staining, Immunofluorescence, Control, Western Blot, Expressing, Quantitative RT-PCR, Two Tailed Test

Journal: Cells

Article Title: Interaction between Chromodomain Y-like Protein and Androgen Receptor Signaling in Sertoli Cells Accounts for Spermatogenesis

doi: 10.3390/cells13100851

Figure Lengend Snippet: AR and CDYL regulate downstream AR-targeted genes. ChIP and luciferase assays demonstrate the CDYL ( A ) or TNP1 ( B ) genes in the presence of AR binding sequences. Schematic of a putative binding site for the transcription factor, androgen-responsive elements (ARE) sequences on the CDYL or TNP1 promoter. Promoter occupancy of AR on CDYL or TNP1 promoter by ChIP. CDYL or TNP1 upstream regions starting at position −3000 were introduced into the pGL3-Basic plasmid before the luciferase reporter gene. The siRNA-mediated knockdown ( C ) or overexpression ( D ) of AR or CDYL substantially changed the CDYL or TNP1 promoter activity in TM4 cells through dual-luciferase assays. * p ˂ 0.05 for one-way ANOVA was significant compared with the control. ( n ≥ 3) in each group, and error bars represent ± SD.

Article Snippet: The TM4 mouse Sertoli cell line (BCRC-60254) was procured from the Bioresource Collection and Research Center (BCRC) in Hsinchu, Taiwan.

Techniques: Luciferase, Binding Assay, Plasmid Preparation, Knockdown, Over Expression, Activity Assay, Control

Journal: Cells

Article Title: Interaction between Chromodomain Y-like Protein and Androgen Receptor Signaling in Sertoli Cells Accounts for Spermatogenesis

doi: 10.3390/cells13100851

Figure Lengend Snippet: mRNA expression of AR , CDYL , and TNP1 association by CDYL rescue in Sertoli cells. mRNA expression of AR , CDYL , and TNP1 association among control, AR siRNA-treated, and CDYL siRNA-treated groups in TM4 cells by non-treat and CDYL rescue. mRNA expression of AR ( A , B ), CDYL ( C , D ), or TNP1 ( E , F ) were detected after being transiently transfected with control, AR , or CDYL siRNA-treated ( A , C , E ). Moreover, these three groups were treated with 200 ng CDYL recombinant protein ( B , D , F ) for 24 h in TM4 cells. * p ˂ 0.05 for one-way ANOVA was significant compared with the control. ( n ≥ 3) in each group, and error bars represent ± SD.

Article Snippet: The TM4 mouse Sertoli cell line (BCRC-60254) was procured from the Bioresource Collection and Research Center (BCRC) in Hsinchu, Taiwan.

Techniques: Expressing, Control, Transfection, Recombinant

Journal: Cells

Article Title: Interaction between Chromodomain Y-like Protein and Androgen Receptor Signaling in Sertoli Cells Accounts for Spermatogenesis

doi: 10.3390/cells13100851

Figure Lengend Snippet: Decreased expression of CDYL in the human testes from patients with azoospermia . Immunohistochemistry indicated the expression of AR and CDYL protein in testicular histology from obstructive azoospermia (active spermatogenesis) and non-obstructive azoospermia (defective spermatogenesis), including Sertoli cell-only syndrome (SCOS) and maturation arrest (halted at the primary spermatocyte stage). Bar = 50 μm. The AR and CDYL signals in patients with normal group ( A , D ), Sertoli cell-only syndrome ( B , E ), and maturation arrest ( C , F ) ( n = 1 patient in every group). S: Sertoli cell; L: Leydig cell; M: myoid cell; SP: spermatids.

Article Snippet: The TM4 mouse Sertoli cell line (BCRC-60254) was procured from the Bioresource Collection and Research Center (BCRC) in Hsinchu, Taiwan.

Techniques: Expressing, Immunohistochemistry