Journal: JACC: Basic to Translational Science

Article Title: Kinetics and Signal Activation Properties of Circulating Factor(s) From Healthy Volunteers Undergoing Remote Ischemic Pre-Conditioning

doi: 10.1016/j.jacbts.2016.01.007

Figure Lengend Snippet: STAT3 Inhibition With Stattic Abolished the Protection by Dialysates of Human Plasma Obtained After RIPC in Murine Myocardium (A) Effect of Stattic (signal transducer and activator of transcription 3 [STAT3] inhibitor) on infarct size and (B) STAT3 phosphorylation in murine myocardium perfused with dialysates of human plasma obtained before (baseline) as well as 30 min and 6 days after RIPC for n = 7 each; *p < 0.05 versus without Stattic. d = day; LV+RV = left and right ventricle; RIPC = remote ischemic pre-conditioning; Tyr = tyrosine.

Article Snippet: Further investigation of the SAFE pathway was performed by inhibition of STAT3 using Stattic (Stattic, Tocris bioscience, Bristol, England) , .

Techniques: Inhibition, Clinical Proteomics, Phospho-proteomics

Journal: Journal of advanced research

Article Title: Lactobacillus johnsonii-derived extracellular vesicles carrying GAPDH protect against ulcerative colitis through modulating macrophage polarization.

doi: 10.1016/j.jare.2025.06.035

Figure Lengend Snippet: Fig. 6. A STAT3 inhibitor alleviates DSS-induced colitis by modulating macrophage phenotype (A) Experiment design of the treatment of mice with DSS and Stattic. (B) Body weight changes. (C) Disease activity indexes. (D) Colonic lengths. (E) Colonic histopathological scores. (F, G) The proportions of CD11c+ cells (F) and CD206+ cells (G) in colon tissues were determined using immunohistochemistry. (H) The serum and colon tissue concentrations of IL-1b, IL-6, TNF-a, IL-10, DAO, and D-LA. (I) Protein expression and analysis of iNOS, Arg1, ZO-1, occludin, and claudin-1 in colon tissues. (J) Experiment design of the treatment of RAW264.7 cells with LPS and Stattic. (K) Levels of iNOS, CD11c, Arg1, and CD206 mRNA in RAW264.7 cells. (L) Experiment design of the RAW264.7 and CT-26 cell co-culture system. (M) Levels of ZO-1, occludin, and claudin-1 mRNA in co- cultured CT-26 cells. n = 8 for A-H; n = 3 for I; n = 4 for J-M. Data are expressed as the means ± SEM (B-I, K and M) and one-way ANOVA was performed, followed by Tukey’s test (B-I, K and M). *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Mice in the DSS + GAPDH group received intraperitoneal injections of 10 mg/kg GAPDH (G2267, Sigma-Aldrich, USA) every 2 d. Mice in the DSS + Stattic group received intraperitoneal injections of 10 mg/kg Stattic (S7024, Selleck, USA) every 2 d. Mice in the DSS + WT-mEVs and DSS + Stattic-mEVs groups were injected intraperitoneally with 20 lg WT-mEVs or Stattic-mEVs every 2 d. Mice in the DSS + Stattic-mEVs-NA and DSS + Stattic-mEVs-PK groups were injected intraperitoneally with 20 lg native or protease-treated Stattic-mEVs every 2 d.

Techniques: Activity Assay, Immunohistochemistry, Expressing, Co-Culture Assay, Cell Culture

Journal: Journal of advanced research

Article Title: Lactobacillus johnsonii-derived extracellular vesicles carrying GAPDH protect against ulcerative colitis through modulating macrophage polarization.

doi: 10.1016/j.jare.2025.06.035

Figure Lengend Snippet: Fig. 7. EVs of macrophage lacking STAT3 enhance the intestinal barrier in colitis mice by inhibiting the TLR signaling pathway (A) TEM of isolated EVs derived from wild-type macrophage and Stattic-treated macrophage. (B) Size distribution of EVs derived from wild-type macrophage and Stattic-treated macrophage analyzed by NTA. (C) Experiment design of the treatment of mice with DSS and Stattic-mEVs. (D) Body weight change in mice. (E) Disease activity index in mice. (F) Colonic picture and length analysis in mice. (G) Colonic morphology and histopathological score in mice. (H) The serum and colon tissue concentration of IL-1b, IL-6, TNF-a, IL-10, DAO, and D-LA in mice. (I) Colonic protein expression and analysis of ZO-1, Occludin, Claudin-1, TLR4, MyD88, and p-NF-jB p65 in mice. n = 8 for C–H; n = 3 for I. Data are expressed as the means ± SEM (D-I) and one-way ANOVA was performed, followed by Tukey’s test (D-I). *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Mice in the DSS + GAPDH group received intraperitoneal injections of 10 mg/kg GAPDH (G2267, Sigma-Aldrich, USA) every 2 d. Mice in the DSS + Stattic group received intraperitoneal injections of 10 mg/kg Stattic (S7024, Selleck, USA) every 2 d. Mice in the DSS + WT-mEVs and DSS + Stattic-mEVs groups were injected intraperitoneally with 20 lg WT-mEVs or Stattic-mEVs every 2 d. Mice in the DSS + Stattic-mEVs-NA and DSS + Stattic-mEVs-PK groups were injected intraperitoneally with 20 lg native or protease-treated Stattic-mEVs every 2 d.

Techniques: Isolation, Derivative Assay, Activity Assay, Concentration Assay, Expressing

Journal: Cell reports

Article Title: VISTA promotes the metabolism and differentiation of myeloid-derived suppressor cells by STAT3 and polyamine-dependent mechanisms

doi: 10.1016/j.celrep.2023.113661

Figure Lengend Snippet: WT and Vsir −/− BM progenitor cells were cultured with GM-CSF (10 ng/mL) and IL-6 (10 ng/mL) for 4 days either in normoxia or exposed to hypoxia during the last 24 h. M-MDSCs (Ly6C + Ly6G neg CD11c neg ) were purified and cocultured with OT1 splenocytes (50,000) at the indicated ratios in the presence of the ovalbumin peptide (SIINFEKL, 100 μg/mL). Expanded OT1 T cells were enumerated by flow cytometry after 72 h of stimulation. (A) Normoxic M-MDSCs and OT1 cells were co-cultured under normoxia for 72 h (n = 4 replicates). (B) Hypoxic M-MDSCs and OT1 cells were continuously co-cultured under hypoxia for 72 h (n = 4 replicates). (c) Hypoxic M-MDSCs were co-cultured with OT1 cells under normoxia for 72 h (n = 4 replicates). (D and E) Protein expression of Arg1 and iNOS in normoxic or hypoxic M-MDSCs after 4-day culture was examined by flow cytometry (n = 3 replicates) and by western blotting. (F) Hypoxic WT and Vsir −/− M-MDSCs were rested for 8 h in medium without any cytokines and restimulated with IL-6. Cells were lysed at the indicated time points. Phosphorylated and total levels of STAT3 were detected by western blotting. (G) Hypoxic WT and Vsir −/− M-MDSCs were rested for 8 h and restimulated with GM-CSF. Cell lysates were generated and examined for phosphorylated and total levels of STAT5 and ERK1/2. The ratios of phosphorylated vs. total proteins were quantified using ImageJ. (H and I) Vsir −/− BM progenitor cells were transduced with a retrovirus expressing the mutant STAT3 proteins Y705E.GFP and S727D.GFP as indicated. Cells were expanded in GM-CSF and IL-6 for 4 days, and the expression of Arg1 was examined by flow cytometry. GFP neg cells that did not express the mutant STAT3 are shown as parallel negative controls. All data were presented as mean ± SEM. * p < 0.05; ** p < 0.025; *** p < 0.005; **** p < 0.0001. All experiments were repeated at least three times, and representative results are shown.

Article Snippet: STAT3 inhibitor (Stattic) , Tocris Bioscience Inc , 2798.

Techniques: Cell Culture, Purification, Flow Cytometry, Expressing, Western Blot, Generated, Transduction, Mutagenesis

Journal: Cell reports

Article Title: VISTA promotes the metabolism and differentiation of myeloid-derived suppressor cells by STAT3 and polyamine-dependent mechanisms

doi: 10.1016/j.celrep.2023.113661

Figure Lengend Snippet: (A and B) WT and Vsir −/− BM-derived MDSCs were cultured with GM-CSF and IL-6 for 4 days either in normoxia or exposed to hypoxia during the last 24 h. Total cell lysates were extracted and examined for ODC1 expression (A). Intracellular putrescine levels were quantified by liquid chromatography-tandem mass spectrometry (LC-MS/MS) (B) (n = 4 for normoxia replicates, n = 3 for hypoxia replicates). (c) Hypoxic WT and Vsir −/− MDSCs were cultured as in (A) in the presence of putrescine (800 μM) or vehicle control. Cells were rested for 8 h before being restimulated with IL-6. Total cell lysates were examined by western blotting. The ratios of phosphorylated-STAT3 Y705 and STAT3 S727 vs. total STAT3 were quantified using ImageJ. (D) WT BM-derived MDSCs were cultured as in (A) in the presence of DFMO (400 μM) or vehicle control for 4 days. Total cell lysates were generated, and levels of pSTAT3 Y705 and total STAT3 were examined by western blotting. (E) WT BM progenitors (n = 3) were cultured for 4 days as in (A). On day +1, CK2 inhibitor (20 μM), putrescine (800 μM), vehicle control, or combined drugs were added to the culture medium and continuously cultured for another 3 days. During the last 24 h, cells were treated with hypoxia before flow analysis to detect Arg1 expression. (F) WT and Vsir −/− BM-MDSCs (n = 3) were cultured for 4 days as in (A). On day +1, STAT3 inhibitor (2 mM), putrescine (1 mM), vehicle control, or combined drugs were added to the culture medium. Cells were treated with hypoxia during the last 24 h before flow analysis. All experiments were repeated at least three times, and representative results are shown. (G and H) WT and Vsir −/− BM-MDSCs were cultured for 4 days under normoxia or exposed to hypoxia during the last 24 h. Mitochondrial respiration of normoxic (G) and hypoxic MDSCs (H) was examined using the Seahorse XF Cell Mito stress test. The OCR and ECAR were recorded simultaneously. (I) Basal OCR, maximal OCR, SRC, and ECAR are summarized. (J) The bioenergetic profiles of WT and Vsir −/− MDSCs were plotted based on basal OCR and ECAR levels. (K and L) To rescue the defective mitochondrial function in Vsir −/− MDSCs, putrescine (800 μM) was added to the culture medium on day +1. Cells were cultured for a total of 4 days and treated with hypoxia during the last 24 h. M-MDSCs were purified and examined using the Mito stress test. OCR was recorded (K). Basal OCR, maximal OCR, and SRC are summarized (L). (M and N) Total cell lysates were extracted, and the expression of ETC complex V and Arg1 was examined. β-Actin was used as the loading control. The ratios of each protein vs. β-actin were quantified using ImageJ. All experiments were repeated at least three times, and representative results are shown. Statistical analysis: n = 8 (WT, normoxia), 9 (KO, normoxia), 8 (WT, hypoxia), and 10 (KO, hypoxia) replicates (A–C); n = 5 (WT control), 4 (WT putrescine), 14 (KO, control), and 8 (KO putrescine) replicates (D). All data were presented as mean ± SEM. * p < 0.05; ** p < 0.025; *** p < 0.005; **** p < 0.0001.

Article Snippet: STAT3 inhibitor (Stattic) , Tocris Bioscience Inc , 2798.

Techniques: Derivative Assay, Cell Culture, Expressing, Liquid Chromatography, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy, Control, Western Blot, Generated, Purification

Journal: Cell reports

Article Title: VISTA promotes the metabolism and differentiation of myeloid-derived suppressor cells by STAT3 and polyamine-dependent mechanisms

doi: 10.1016/j.celrep.2023.113661

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: STAT3 inhibitor (Stattic) , Tocris Bioscience Inc , 2798.

Techniques: Virus, Western Blot, Recombinant, Staining, cDNA Synthesis, SYBR Green Assay, Isolation, Mutagenesis

Journal: Molecular Cancer

Article Title: ISL-1 is overexpressed in non-Hodgkin lymphoma and promotes lymphoma cell proliferation by forming a p-STAT3/p-c-Jun/ISL-1 complex

doi: 10.1186/1476-4598-13-181

Figure Lengend Snippet: JNK or JAK/STAT signaling inhibitors inhibit NHL cells proliferation through down-regulating ISL-1 expression. (A) The relative proliferation rate of lymphoma cell lines were measured using CCK-8 analysis after treated with JNK signaling pathway inhibitor (SP60012, 10 μM) or JAK/STAT signaling pathway inhibitor (STATTIC, 6 μM). The cell treated with DMSO were used as the control. (B to C) The effect of SP600125 (10 μM) and STATTIC (6 μM) on ISL-1 and c-Myc expression at both mRNA (B) and protein levels (C) were analyzed by real-time RT-PCR and Western blot. The cells treated with DMSO at different time point were used as the corresponding control. (D) The luciferase activity of c-Myc-luc (wide type or M1) was measured in Ly3 cells with or without ISL-1 transcfection after treated with 10 μM SP600125 or 6 μM STATTIC for 24 h. (E) The growth inhibition of Ly3 cells with or without ISL-1 transcfection was measured by CCK-8 analysis after treated with 10 μM SP600125 or 6 μM STATTIC for 24 h. The data represent three independent experiments. Each bar represents mean ± SD. p values were calculated using a Student t -test (* p < 0.05, ** p < 0.01, # p < 0.05, ## p < 0.01 vs. each control).

Article Snippet: STATTIC (sc-202818, Santa Cruz Biotechnology, Santa Cruz, CA, USA), SP600125 (s5567) and Anisomycin (A9789) purchased from Sigma (Louis, MO, USA) were dissolved in 100% dimethyl sulfoxide to prepare a 40 mM stock, 20 mM stock and 15 mg/ml stock respectively, and stored at -20°C.

Techniques: Expressing, CCK-8 Assay, Control, Quantitative RT-PCR, Western Blot, Luciferase, Activity Assay, Inhibition

Journal: Molecular Cancer

Article Title: ISL-1 is overexpressed in non-Hodgkin lymphoma and promotes lymphoma cell proliferation by forming a p-STAT3/p-c-Jun/ISL-1 complex

doi: 10.1186/1476-4598-13-181

Figure Lengend Snippet: p-STAT3/p-c-Jun/ ISL-1 forms a transcriptional complex and binds directly to ISL-1 promoter. (A) Consensus binding sites for p-STAT3 and p-c-Jun on the ISL-1 promoter were analyzed by Matinspector software. (B) The luciferase activity of ISL-1-luc was analyzed by luciferase reporter assay in Ly3 cells after treated with IL-6 (4 ng/ml), STATTIC (6 μM), Anisomycin (15 ng/ml) or SP600125 (10 μM) for 24 h. (C) ChIP assay was performed with anti-p-STAT3 Ab (left panel) or anti-p-c-Jun Ab (right panel) for immunoprecipitation using chromatin harvested from Ly3 cells. The DNA extractions were amplified using the primers that cover the p-STAT3 (primers 2) or p-c-Jun (primers 4) binding sites, or control primers (primers 1, 3) on the ISL-1 promoter by real-time PCR with normal IgG as a control. (D) Co-IP assay was performed in Ly3 for the transcriptional complex recruited on the ISL-1 promoter. (E) ChIP-re-IP assay was performed first with anti-ISL-1 Ab or rabbit IgG Ab and then with anti-p-STAT3, anti-p-c-Jun or IgG Abs for immunoprecipitation using chromatin harvested from Ly3 cells. (F) The transcriptional activity of ISL-1 on ISL-1-luc was analyzed in Ly3 cells by luciferase reporter assay. The data represent three independent experiments. Each bar represents mean ± SD. p values were calculated using a Student t -test (* p < 0.05, ** p < 0.01 vs. the control).

Article Snippet: STATTIC (sc-202818, Santa Cruz Biotechnology, Santa Cruz, CA, USA), SP600125 (s5567) and Anisomycin (A9789) purchased from Sigma (Louis, MO, USA) were dissolved in 100% dimethyl sulfoxide to prepare a 40 mM stock, 20 mM stock and 15 mg/ml stock respectively, and stored at -20°C.

Techniques: Binding Assay, Software, Luciferase, Activity Assay, Reporter Assay, Immunoprecipitation, Amplification, Control, Real-time Polymerase Chain Reaction, Co-Immunoprecipitation Assay

Journal: Journal of experimental & clinical cancer research : CR

Article Title: Intravesical instillation-based mTOR-STAT3 dual targeting for bladder cancer treatment.

doi: 10.1186/s13046-024-03088-7

Figure Lengend Snippet: Fig. 2 Efficacy of combined suppression between mTOR and STAT3. A To evaluate the synergistic effect, the viabilities of 253 J-BV and RT-4 cells were measured following individual or combined treatment with mTOR and STAT3 siRNAs (+ : siRNA 50 nM, + + : siRNA 100 nM). B Viability of cisplatin-resistant 253 J-BV cells in the presence of cisplatin (10 μM) with siRNA treatment (+ : siRNA 50 nM, + + : siRNA 100 nM). C Western blotting to evaluate the downstream targets of mTOR and STAT3. Downstream target molecules were evaluated after treatment with Torin1 and STATTIC in 253 J-BV cells (Torin-1; + : 1 μM, + + : 2 μM, STATTIC; + : 5 μM, + + : 10 μM). D RT-4 cells were transfected with mTOR and STAT3 siRNAs. Next, mTOR- and STAT3-related molecules were analyzed (+ : siRNA 50 nM, + + : siRNA 100 nM). E The 253 J-BV and A549 cell lines were subjected to transfection procedures with siRNAs targeted at mTOR and STAT3. Subsequent to this manipulation, a detailed analysis was conducted focusing on molecular entities related to both mTORC1 and mTORC2 complexes (+ : siRNA 50 nM, + + : siRNA 100 nM) (for statistics, two-tailed t-test for A and B)

Article Snippet: The Torin-1 mTOR inhibitor (Cat. no. 14379 s; Cell Signaling Inc., Massachusetts, US) and STATTIC STAT3 inhibitor (Cat. no. 97598 s; Cell Signaling Inc.) were diluted in dimethyl sulfoxide (DMSO, Cat. no. D2650; Sigma Inc., Mijuri, US) for treatment.

Techniques: Western Blot, Transfection, Two Tailed Test

Journal: Journal of experimental & clinical cancer research : CR

Article Title: Intravesical instillation-based mTOR-STAT3 dual targeting for bladder cancer treatment.

doi: 10.1186/s13046-024-03088-7

Figure Lengend Snippet: Fig. 3 Balanced knockdown strategy for mTOR/STAT3 via an shRNA-mediated expression system. A Graphic illustration of the structure of the bispecific shRNA (bs_shRNA) compared with that of the conventional tandem shRNA system (t_shRNA). This system allows the encoding of two target sequences in a short coding length and decreases the off-target effect. B qPCR detection of mTOR and STAT3 expression of 253 J-BV for knock-down efficacy of t_shRNA and bs_shRNA in 253 J-BV transfected with t_shRNA and bs_shRNA. C The number of shRNA production of t_shRNA and bs_shRNA in 253 J-BV at a single cell level. Relative quantification is used through qPCR to calculate the copy number of the shRNA product. Refer to copy number calculation of shRNA in methods for detail method. D qPCR analysis at the single-cell level to compare the efficacy of knock-down between t_shRNA and bs_shRNA in each 253 J-BV transfected plasmid vector. Refer to RNA preparation and qPCR from a single cell in methods for detail method. (For statistics, two-tailed t-test for B and C, NS = non-significant)

Article Snippet: The Torin-1 mTOR inhibitor (Cat. no. 14379 s; Cell Signaling Inc., Massachusetts, US) and STATTIC STAT3 inhibitor (Cat. no. 97598 s; Cell Signaling Inc.) were diluted in dimethyl sulfoxide (DMSO, Cat. no. D2650; Sigma Inc., Mijuri, US) for treatment.

Techniques: Knockdown, shRNA, Expressing, Transfection, Quantitative Proteomics, Plasmid Preparation, Two Tailed Test

Journal: Journal of experimental & clinical cancer research : CR

Article Title: Intravesical instillation-based mTOR-STAT3 dual targeting for bladder cancer treatment.

doi: 10.1186/s13046-024-03088-7

Figure Lengend Snippet: Fig. 4 Incorporation of bispecific shRNA into replication-competent adenovirus. A Genetic construction of bispecific shRNA (bs_shRNA)-expressing adenovirus (BSV). The human telomerase promoter was encoded in the front of E1A-IRES-E1B, and the U6 promoter was used for the shRNA expression in E3. In CV, the shRNA coding region was replaced by a GFP-coding sequence. Refer to preparation of replication-competent adenovirus in methods for detail method. B Normal cells (PrEC and HUEpC) and cancer cells (C4-2B and 253 J-BV) were infected by 20 MOI of CV for 72 h. C Viral vector concentration (MOI)-based cell viability test: HUEpC and 253 J-BV cells were treated with 5 MOI of CV and BSV for 72 h. D Suppression of the expression of mTOR and STAT3 as indicated by real-time PCR. For this analysis, 253 J-BV cells were treated with 5 MOI CV and BSV for 72 h. E Western blotting revealing the changes between BSV- and CV-induced mTOR and STAT3 downregulation following the treatment of 253 J-BV cells with 5 MOI CV and BSV for 72 h. F, G Viral vector concentration (MOI)-based cell viability test using crystal violet staining (F) and cell viability assay (G). The 253 J-BV cells were treated with viruses for 72 h in a concentration-dependent manner (for statistics, two-tailed t-test for C, D)

Article Snippet: The Torin-1 mTOR inhibitor (Cat. no. 14379 s; Cell Signaling Inc., Massachusetts, US) and STATTIC STAT3 inhibitor (Cat. no. 97598 s; Cell Signaling Inc.) were diluted in dimethyl sulfoxide (DMSO, Cat. no. D2650; Sigma Inc., Mijuri, US) for treatment.

Techniques: shRNA, Expressing, Sequencing, Infection, Plasmid Preparation, Concentration Assay, Real-time Polymerase Chain Reaction, Western Blot, Staining, Viability Assay, Two Tailed Test

Journal: International Journal of Nanomedicine

Article Title: Chitosan-Coated-PLGA Nanoparticles Enhance the Antitumor and Antimigration Activity of Stattic – A STAT3 Dimerization Blocker

doi: 10.2147/IJN.S337093

Figure Lengend Snippet: Stattic released in PBS and plasma. Cumulative percentage of Stattic released from S@PLGA NPs and S@C-PLGA NPs in different solution at 0–48 h ( A ) and 0–2 h ( B ). Data represent mean ± SEM from three independent experiments; ****Indicate p < 0.0001, as assessed by two-way ANOVA with Tukey’s post-hoc test.

Article Snippet: In this study, the changes in the antimigration properties of Stattic before and after its entrapment into the C-PLGA NPs (ie, Stattic vs S@C-PLGA) were assessed using human metastatic MDA-MB-231 cells and murine metastatic 4T1 cells., Stattic entrapped in the C-PLGA nanocarrier exhibited better in vitro antimigration effects than S@PLGA and Stattic, whereby S@C-PLGA at 1 μM Stattic equivalent concentration suppressed the migration of MDA-MB-231 ( ) and 4T1 ( ) cells by 65.1% and 47.3%, respectively ( ).

Techniques: Clinical Proteomics

Journal: International Journal of Nanomedicine

Article Title: Chitosan-Coated-PLGA Nanoparticles Enhance the Antitumor and Antimigration Activity of Stattic – A STAT3 Dimerization Blocker

doi: 10.2147/IJN.S337093

Figure Lengend Snippet: In vitro anti-migration properties of Stattic, S@PLGA and S@C-PLGA NPs in Scratch Assay. Microscopic images of the Scratch Assay on MDA-MB-231 ( A ) and 4T1 ( B ) at different time points. ( C ) The normalized migration rate of MDA-MB-231 and 4T1 cells after being treated with Stattic, S@PLGA and S@C-PLGA NPs at 1 µM Stattic equivalent concentration for 24 h. Data represent mean ± SEM from three independent experiments; ** and ****Indicate p < 0.01, 0.0001, respectively, as assessed by two-way ANOVA with Tukey’s post-hoc test.

Article Snippet: In this study, the changes in the antimigration properties of Stattic before and after its entrapment into the C-PLGA NPs (ie, Stattic vs S@C-PLGA) were assessed using human metastatic MDA-MB-231 cells and murine metastatic 4T1 cells., Stattic entrapped in the C-PLGA nanocarrier exhibited better in vitro antimigration effects than S@PLGA and Stattic, whereby S@C-PLGA at 1 μM Stattic equivalent concentration suppressed the migration of MDA-MB-231 ( ) and 4T1 ( ) cells by 65.1% and 47.3%, respectively ( ).

Techniques: In Vitro, Migration, Wound Healing Assay, Concentration Assay

Journal: International Journal of Nanomedicine

Article Title: Chitosan-Coated-PLGA Nanoparticles Enhance the Antitumor and Antimigration Activity of Stattic – A STAT3 Dimerization Blocker

doi: 10.2147/IJN.S337093

Figure Lengend Snippet: In vitro anti-migration properties of Stattic, S@PLGA and S@C-PLGA NPs in transwell assay. Microscopic images of the transwell migration assay in MDA-MB-231 and 4T1 after treated with 1 μM of Stattic, S@PLGA NPs and S@C-PLGA NPs. The percentage of migrated cells relative to control were quantified using ImageJ software version 2.0. Data represent mean ± SEM from three independent experiments; ** and ****Indicate p < 0.01, 0.0001, respectively, as assessed by two-way ANOVA with Tukey’s post-hoc test.

Article Snippet: In this study, the changes in the antimigration properties of Stattic before and after its entrapment into the C-PLGA NPs (ie, Stattic vs S@C-PLGA) were assessed using human metastatic MDA-MB-231 cells and murine metastatic 4T1 cells., Stattic entrapped in the C-PLGA nanocarrier exhibited better in vitro antimigration effects than S@PLGA and Stattic, whereby S@C-PLGA at 1 μM Stattic equivalent concentration suppressed the migration of MDA-MB-231 ( ) and 4T1 ( ) cells by 65.1% and 47.3%, respectively ( ).

Techniques: In Vitro, Migration, Transwell Assay, Transwell Migration Assay, Control, Software

Journal: International Journal of Nanomedicine

Article Title: Chitosan-Coated-PLGA Nanoparticles Enhance the Antitumor and Antimigration Activity of Stattic – A STAT3 Dimerization Blocker

doi: 10.2147/IJN.S337093

Figure Lengend Snippet: ( A ) In vivo toxicity profiling and antitumor efficacy study of Stattic and S@C-PLGA NPs. In vivo toxicity profile of Stattic and S@C-PLGA NPs in 4T1 tumor-bearing mice. Data represents the mean body weights ± SEM (n = 3) for each group. ( B ) In vivo antitumor efficacy of Stattic and S@C-PLGA NPs in 4T1 tumor-bearing mice. S@C-PLGA NPs (24 mg Stattic eqv./kg) showed greater suppression in 4T1 tumor growth compared to Stattic (24 mg/kg) and saline. The day of treatment was indicated as black arrows. Data represents the mean tumor volume ± SEM (n = 4) for each group; ** and **** indicate p < 0.01, 0.0001, respectively, as assessed by one-way ANOVA with Dunnett’s post-hoc test.

Article Snippet: In this study, the changes in the antimigration properties of Stattic before and after its entrapment into the C-PLGA NPs (ie, Stattic vs S@C-PLGA) were assessed using human metastatic MDA-MB-231 cells and murine metastatic 4T1 cells., Stattic entrapped in the C-PLGA nanocarrier exhibited better in vitro antimigration effects than S@PLGA and Stattic, whereby S@C-PLGA at 1 μM Stattic equivalent concentration suppressed the migration of MDA-MB-231 ( ) and 4T1 ( ) cells by 65.1% and 47.3%, respectively ( ).

Techniques: In Vivo, Saline

Journal: International Journal of Nanomedicine

Article Title: Chitosan-Coated-PLGA Nanoparticles Enhance the Antitumor and Antimigration Activity of Stattic – A STAT3 Dimerization Blocker

doi: 10.2147/IJN.S337093

Figure Lengend Snippet: In vivo antimetastatic effects of S@C-PLGA NPs. Histopathological analysis and quantification of metastatic foci for lung, liver, and lymph node of untreated control mice ( A ) and 4T1 tumor-bearing mice treated with multiple doses of saline ( B ), Stattic (24 mg/kg) ( C ) and S@C-PLGA NPs (24 mg Stattic eqv./kg) ( D ). No metastatic foci were observed in heart, kidney and spleen. Quantification of number of metastatic foci found in lung ( E ) and liver ( F ) by observing 5 and 8 images in lung and liver, respectively, for each mouse in all the treatment groups. Average sizes of metastatic foci found in lung ( G ) and liver ( H ) for each treatment group. Data represent the mean number of metastatic foci ± SEM (n=4); *, **, *** and ****Indicate p <0.05, 0.01, 0.001 and 0.0001, respectively as assessed by One-way ANOVA with Dunnett’s post-hoc test. Yellow arrows indicate metastatic foci.

Article Snippet: In this study, the changes in the antimigration properties of Stattic before and after its entrapment into the C-PLGA NPs (ie, Stattic vs S@C-PLGA) were assessed using human metastatic MDA-MB-231 cells and murine metastatic 4T1 cells., Stattic entrapped in the C-PLGA nanocarrier exhibited better in vitro antimigration effects than S@PLGA and Stattic, whereby S@C-PLGA at 1 μM Stattic equivalent concentration suppressed the migration of MDA-MB-231 ( ) and 4T1 ( ) cells by 65.1% and 47.3%, respectively ( ).

Techniques: In Vivo, Control, Saline

Journal: Cancers

Article Title: Stromal POSTN Enhances Motility of Both Cancer and Stromal Cells and Predicts Poor Survival in Colorectal Cancer

doi: 10.3390/cancers15030606

Figure Lengend Snippet: POSTN enhanced the motility but suppressed the proliferation of NIH/3T3 cells. ( a , b ) Exogenous POSTN enhanced the migration of NIH/3T3 cells. Bar = 100 µm. Assays were performed in quadruplicate. Data are shown as mean ± S.D. **, p < 0.01. ( c ) POSTN suppressed the cellular proliferation of NIH/3T3 cells. Assays were performed in triplicate. Data are shown as mean ± S.D. *, p < 0.05. ( d ) Immunoblot analysis showing upregulated STAT3 and P-STAT3 in NIH/3T3 cells. Exogeneous POSTN upregulated P-STAT3 at S727. Co-culture with CRC cells upregulated basal STAT3 and P-STAT3 at Y705 in NIH/3T3. ( e ) Stattic, a selective inhibitor for STAT3, significantly downregulated the migration of NIH/3T3 cells at 10 µM. Assays were performed in quadruplicate. Data are shown as mean ± S.D. **, p < 0.01. The uncropped bolts are shown in .

Article Snippet: Stattic, a selective inhibitor for STAT3, was obtained from FUJIFILM Wako Pure Chemical Corporation (Osaka, Japan).

Techniques: Migration, Western Blot, Co-Culture Assay