s3i 201  (Millipore)

Journal: Neural Regeneration Research

Article Title: Ruxolitinib improves the inflammatory microenvironment, restores glutamate homeostasis, and promotes functional recovery after spinal cord injury

doi: 10.4103/NRR.NRR-D-23-01863

Figure Lengend Snippet: RUX restores EAAT2 loss in astrocytes by inhibiting the activation of STAT3 in vitro. (A) Representative western blotting of p-JAK2, JAK2, p-STAT3 and STAT3 expression of astrocytes in each group pre-treated with RUX (0.2, 0.5 and 1 µM) ( n = 3 per group). (B) Quantitative analysis of p-JAK2/JAK2 and p-STAT3/STAT3 level in each group. (C) Immunocytochemistry of GFAP (green, Alexa Fluor 488) and p-STAT3 (red, Alexa Fluor 594) in primary mouse astrocytes. DAPI (blue) was used to stain nuclei. The A1IM group exhibited higher nuclear p-STAT3 expression compared with the Control group, whereas the RUX group displayed decreased nuclear p-STAT3 expression compared with the A1IM group. Scale bar: 40 µm. (D) Quantitative results of relative p-STAT3 intensity. (E) Representative western blotting of STAT3 expression of astrocytes pre-treated with S3I-201 or vehicle. (F) Quantitative results of relative STAT3 expression level. (G) Primary mouse astrocytes were pretreated with S3I-201 (50 μM) or RUX for 1 hour and then exposed to LPS (1 µg/mL) or PBS for 5 hours. Western blotting was conducted to assess EAAT2 expression level, followed by quantitative analysis of EAAT2 protein expression (H). (I) Glutamate (100 nM) was introduced into each culture medium, and the relative glutamate uptake of astrocytes in each group was measured. Data are normalized to the control group. Data are presented as the mean ± SD ( n = 3 per group). ## P < 0.01, vs. control group; * P < 0.05, ** P < 0.01, vs. A1IM group (one-way analysis of variance followed by Tukey’s post hoc test). A1IM: A1-like astrocyte induction medium; Ctrl: control; DAPI: 4′,6-diamidino-2-phenylindole; EAAT2: excitatory amino acid transporter 2; GFAP: glial fibrillary acidic protein; JAK2: Janus kinase 2; ns: not significant; p-JAK2: phosphorylated JAK2; p-STAT3: phosphorylated STAT3; RUX: ruxolitinib; S3I-201: a STAT3 inhibitor; STAT3: signal transducer and activator of transcription 3.

Article Snippet: In experiments, cells were starved in DMEM for 1 hour before being treated with RUX (0.2, 0.5, and 1 µM), S3I-201 (50 µm; Selleck, Houston, TX, USA), or solvent (dimethyl sulfoxide, DMSO).

Techniques: Activation Assay, In Vitro, Western Blot, Expressing, Immunocytochemistry, Staining

Journal: Cancer Cell International

Article Title: Enhanced anti-tumor efficacy of S3I-201 in breast cancer mouse model through Wharton jelly- exosome

doi: 10.1186/s12935-024-03501-3

Figure Lengend Snippet: Determination of S3I-201 loading efficiency into exosomes and evaluation of cell viability. ( A ) 4T1 cells were exposed to 1 µg exosome with different concentrations of WJ-Exo (S3I-201) and free S3I-201 (10, 100, 200, 300, 400, 500 µM) for 48 h in a medium-free environment. Cell viability was measured by MTT assay, and WJ-Exo(S3I-201) showed increased cytotoxicity with an IC50 of 301.4 µM, exceeding the cytotoxicity of free S3I-201 with an IC50 of 337.1 µM. ( B ) The dynamic light scattering (DLS) determined size of WJ-Exo after loading was measured to be 117.3 ± 5.6 nm. ( C ) The loading efficiency of S3I-201, calculated from the HPLC area under the curve and the standard curve, was 42.26%

Article Snippet: The mice then received intraperitoneal injections of DMSO, S3I-201 (56 µg/dose), WJ-Exo (10 µg of exosome) and WJ-Exo (S3I-201) (10 µg of exosome loaded with 56 µg S3I-201/dose) on days 10, 12 and 14.

Techniques: MTT Assay

Journal: Cancer Cell International

Article Title: Enhanced anti-tumor efficacy of S3I-201 in breast cancer mouse model through Wharton jelly- exosome

doi: 10.1186/s12935-024-03501-3

Figure Lengend Snippet: In vitro anti-tumor effects of S3I-201-loaded WJ-Exos. ( A ) Cell viability of 4T1 cells was examined after treatment with DMSO, S3I-201 (IC50: 337.1), WJ-Exo, WJ-Exo control and WJ-Exo(S3I-201, IC50: 301.4 µM) for 24, 48 and 72 h using the MTT assay. WJ-Exo-control means incubation of WJ-Exo with S3I-201 without electroporation to test passive loading of drug to exosomes. ( B ) The viability of healthy cells remained unaffected after 48 h of incubation with different concentrations of free S3I-201. ( C ) 4T1 cells were cultured in a 24-well plate with 5 µg of WJ-Exo (S3I-201) loaded with S3I-201 (IC50: 301.4 µM) and IC50: 337.1 µM of free S3I-201. Flow cytometry was used to evaluate cell apoptosis in each group after 48 h, and ( D ) shows the results of statistical analysis. The data shown are mean ± SD ( n = 3). (* P < 0.05, ** P < 0.001, *** P < 0.03, **** P < 0.0001 vs. the control group) and (# compare between free S3I-201 vs. WJ-Exo(S3I-201)). Statistical significances indicated with * are compared with the DMSO group. Data underwent statistical analysis using One-Way and Two-Way ANOVA followed by Tukey post hoc test for multiple comparisons

Article Snippet: The mice then received intraperitoneal injections of DMSO, S3I-201 (56 µg/dose), WJ-Exo (10 µg of exosome) and WJ-Exo (S3I-201) (10 µg of exosome loaded with 56 µg S3I-201/dose) on days 10, 12 and 14.

Techniques: In Vitro, Control, MTT Assay, Incubation, Electroporation, Cell Culture, Flow Cytometry

Journal: Cancer Cell International

Article Title: Enhanced anti-tumor efficacy of S3I-201 in breast cancer mouse model through Wharton jelly- exosome

doi: 10.1186/s12935-024-03501-3

Figure Lengend Snippet: Effects of WJ-Exo(S3I-201) treatment on gene and protein expression in 4T1 cells. ( A ) 4T1 cells (1 × 10 5 cells/96-well plate) were cultured with 1 µg of WJ-Exo(S3I-201, 301.4 µM), S3I-201 (337.1 µM), 1 µg of WJ-Exo and DMSO for 48-hour. RT-qPCR was used to measure the mRNA expression of Bcl-2, Bax and caspase-3 in 4T1 cells. (* P < 0.05, ** P < 0.001, *** P < 0.03, **** P < 0.0001 vs. control group). Statistical significances indicated with * are compared with the DMSO group. ( B ) 4T1 cells (7 × 10 5 cells/well) were treated with free S3I-201 (337.1 µM), WJ-Exo(S3I-201) (5 µg exosome loaded with 301.4 µM of S3I-201), WJ-Exo and DMSO. Western blot analysis was performed in the study to analyze the expression levels of STAT3, P-STAT3 (Y705), Bcl-2, Bax, and cleaved caspase-3 in 4T1 cells, with β-actin serving as a control for protein loading. Data underwent statistical analysis using One-Way ANOVA followed by Tukey post hoc test for multiple comparisons

Article Snippet: The mice then received intraperitoneal injections of DMSO, S3I-201 (56 µg/dose), WJ-Exo (10 µg of exosome) and WJ-Exo (S3I-201) (10 µg of exosome loaded with 56 µg S3I-201/dose) on days 10, 12 and 14.

Techniques: Expressing, Cell Culture, Quantitative RT-PCR, Control, Western Blot

Journal: Cancer Cell International

Article Title: Enhanced anti-tumor efficacy of S3I-201 in breast cancer mouse model through Wharton jelly- exosome

doi: 10.1186/s12935-024-03501-3

Figure Lengend Snippet: WJ-Exo(S3I-201) inhibits the migration of 4T1 cells. ( A ) A wound healing assay visually shows the initial and final scratch conditions in the 4T1 cell line treated with free S3I-201 (337.1 µM), WJ-Exo(S3I-201) (5 µg exosome loaded with 301.4 µM of S3I-201), WJ-Exo and DMSO. ( B ) Quantitative results for the wound area analysis are presented. (* P < 0.05, ** P < 0.001, *** P < 0.03, **** P < 0.0001 vs. the control group). Data underwent statistical analysis using Two-Way ANOVA followed by Tukey post hoc test for multiple comparisons

Article Snippet: The mice then received intraperitoneal injections of DMSO, S3I-201 (56 µg/dose), WJ-Exo (10 µg of exosome) and WJ-Exo (S3I-201) (10 µg of exosome loaded with 56 µg S3I-201/dose) on days 10, 12 and 14.

Techniques: Migration, Wound Healing Assay, Control

Journal: Cancer Cell International

Article Title: Enhanced anti-tumor efficacy of S3I-201 in breast cancer mouse model through Wharton jelly- exosome

doi: 10.1186/s12935-024-03501-3

Figure Lengend Snippet: Evaluation of the anti-tumor effects of WJ-Exo, S3I-201 and WJ-Exo(S3I-201) in tumor-bearing mice. ( A ) Determination of tumor volume ( n = 5/group) and ( B ) excision of tumors for weight measurement after 21 days of treatment. ( C ) Monitoring of daily body weight changes in each group. ( D ) Determination of survival rates ( n = 3). The mice received intraperitoneal injections of DMSO, S3I-201 (56 µg/dose), WJ-Exo (10 µg of exosome) and WJ-Exo (S3I-201) (10 µg of exosome loaded with 56 µg/dose S3I-201) on days 10, 12 and 14. Values represent mean ± SD ( n = 8; * P < 0.05, ** P < 0.001, *** P < 0.03, **** P < 0.0001 vs. control). Data underwent statistical analysis using One-Way and Two-Way ANOVA followed by Tukey post hoc test for multiple comparisons, along with Kaplan-Meier estimation for survival analysis

Article Snippet: The mice then received intraperitoneal injections of DMSO, S3I-201 (56 µg/dose), WJ-Exo (10 µg of exosome) and WJ-Exo (S3I-201) (10 µg of exosome loaded with 56 µg S3I-201/dose) on days 10, 12 and 14.

Techniques: Control

Journal: Cancer Cell International

Article Title: Enhanced anti-tumor efficacy of S3I-201 in breast cancer mouse model through Wharton jelly- exosome

doi: 10.1186/s12935-024-03501-3

Figure Lengend Snippet: Evaluation of splenocyte proliferation and cytokine levels. ( A ) Splenocyte proliferation: Mouse splenocytes (10 6 cells/ml) were stimulated with phytohemagglutinin (PHA) and tumor lysate for 72 h at 37 °C and 5% CO2. Non-stimulated splenocytes under identical conditions were used as controls. Statistical significances indicated with * are compared with the DMSO group. Statistical comparisons between the S3I-201 group and WJ-Exo (S3I-201) were labeled with ####. ( B-E ) Cytokine Production: Cytokine levels in cells cultured with PHA and tumor lysate were measured. Murine splenocytes (10 6 cells/ml) were stimulated for 72 h at 37 °C in 5% CO 2 . Controls comprised non-stimulated splenocytes under the same conditions. Data, representative of at least three ( n = 3) independent experiments per group, were analyzed via Student’s t-test ( n = 3; * p < 0.05, ** p < 0.001, *** p < 0.03, *** p < 0.0001 vs. control). Data underwent statistical analysis using Two-Way ANOVA followed by Tukey post hoc test for multiple comparisons ( A-E )

Article Snippet: The mice then received intraperitoneal injections of DMSO, S3I-201 (56 µg/dose), WJ-Exo (10 µg of exosome) and WJ-Exo (S3I-201) (10 µg of exosome loaded with 56 µg S3I-201/dose) on days 10, 12 and 14.

Techniques: Labeling, Cell Culture, Control

Journal: Cancer Cell International

Article Title: Enhanced anti-tumor efficacy of S3I-201 in breast cancer mouse model through Wharton jelly- exosome

doi: 10.1186/s12935-024-03501-3

Figure Lengend Snippet: Determination of S3I-201 loading efficiency into exosomes and evaluation of cell viability. ( A ) 4T1 cells were exposed to 1 µg exosome with different concentrations of WJ-Exo (S3I-201) and free S3I-201 (10, 100, 200, 300, 400, 500 µM) for 48 h in a medium-free environment. Cell viability was measured by MTT assay, and WJ-Exo(S3I-201) showed increased cytotoxicity with an IC50 of 301.4 µM, exceeding the cytotoxicity of free S3I-201 with an IC50 of 337.1 µM. ( B ) The dynamic light scattering (DLS) determined size of WJ-Exo after loading was measured to be 117.3 ± 5.6 nm. ( C ) The loading efficiency of S3I-201, calculated from the HPLC area under the curve and the standard curve, was 42.26%

Article Snippet: After 48-hour treatment of 4T1 cells (1 × 10 5 cells/96-well plate) with 1 µg of WJ-Exo(S3I-201, 301.4 µM), S3I-201 (337.1 µM), 1 µg of WJ-Exo and DMSO, total RNA, including miRNAs, was extracted with Trizol reagent (Gibco, Germany).

Techniques: MTT Assay

Journal: Cancer Cell International

Article Title: Enhanced anti-tumor efficacy of S3I-201 in breast cancer mouse model through Wharton jelly- exosome

doi: 10.1186/s12935-024-03501-3

Figure Lengend Snippet: In vitro anti-tumor effects of S3I-201-loaded WJ-Exos. ( A ) Cell viability of 4T1 cells was examined after treatment with DMSO, S3I-201 (IC50: 337.1), WJ-Exo, WJ-Exo control and WJ-Exo(S3I-201, IC50: 301.4 µM) for 24, 48 and 72 h using the MTT assay. WJ-Exo-control means incubation of WJ-Exo with S3I-201 without electroporation to test passive loading of drug to exosomes. ( B ) The viability of healthy cells remained unaffected after 48 h of incubation with different concentrations of free S3I-201. ( C ) 4T1 cells were cultured in a 24-well plate with 5 µg of WJ-Exo (S3I-201) loaded with S3I-201 (IC50: 301.4 µM) and IC50: 337.1 µM of free S3I-201. Flow cytometry was used to evaluate cell apoptosis in each group after 48 h, and ( D ) shows the results of statistical analysis. The data shown are mean ± SD ( n = 3). (* P < 0.05, ** P < 0.001, *** P < 0.03, **** P < 0.0001 vs. the control group) and (# compare between free S3I-201 vs. WJ-Exo(S3I-201)). Statistical significances indicated with * are compared with the DMSO group. Data underwent statistical analysis using One-Way and Two-Way ANOVA followed by Tukey post hoc test for multiple comparisons

Article Snippet: After 48-hour treatment of 4T1 cells (1 × 10 5 cells/96-well plate) with 1 µg of WJ-Exo(S3I-201, 301.4 µM), S3I-201 (337.1 µM), 1 µg of WJ-Exo and DMSO, total RNA, including miRNAs, was extracted with Trizol reagent (Gibco, Germany).

Techniques: In Vitro, Control, MTT Assay, Incubation, Electroporation, Cell Culture, Flow Cytometry

Journal: Cancer Cell International

Article Title: Enhanced anti-tumor efficacy of S3I-201 in breast cancer mouse model through Wharton jelly- exosome

doi: 10.1186/s12935-024-03501-3

Figure Lengend Snippet: Effects of WJ-Exo(S3I-201) treatment on gene and protein expression in 4T1 cells. ( A ) 4T1 cells (1 × 10 5 cells/96-well plate) were cultured with 1 µg of WJ-Exo(S3I-201, 301.4 µM), S3I-201 (337.1 µM), 1 µg of WJ-Exo and DMSO for 48-hour. RT-qPCR was used to measure the mRNA expression of Bcl-2, Bax and caspase-3 in 4T1 cells. (* P < 0.05, ** P < 0.001, *** P < 0.03, **** P < 0.0001 vs. control group). Statistical significances indicated with * are compared with the DMSO group. ( B ) 4T1 cells (7 × 10 5 cells/well) were treated with free S3I-201 (337.1 µM), WJ-Exo(S3I-201) (5 µg exosome loaded with 301.4 µM of S3I-201), WJ-Exo and DMSO. Western blot analysis was performed in the study to analyze the expression levels of STAT3, P-STAT3 (Y705), Bcl-2, Bax, and cleaved caspase-3 in 4T1 cells, with β-actin serving as a control for protein loading. Data underwent statistical analysis using One-Way ANOVA followed by Tukey post hoc test for multiple comparisons

Article Snippet: After 48-hour treatment of 4T1 cells (1 × 10 5 cells/96-well plate) with 1 µg of WJ-Exo(S3I-201, 301.4 µM), S3I-201 (337.1 µM), 1 µg of WJ-Exo and DMSO, total RNA, including miRNAs, was extracted with Trizol reagent (Gibco, Germany).

Techniques: Expressing, Cell Culture, Quantitative RT-PCR, Control, Western Blot

Journal: Cancer Cell International

Article Title: Enhanced anti-tumor efficacy of S3I-201 in breast cancer mouse model through Wharton jelly- exosome

doi: 10.1186/s12935-024-03501-3

Figure Lengend Snippet: WJ-Exo(S3I-201) inhibits the migration of 4T1 cells. ( A ) A wound healing assay visually shows the initial and final scratch conditions in the 4T1 cell line treated with free S3I-201 (337.1 µM), WJ-Exo(S3I-201) (5 µg exosome loaded with 301.4 µM of S3I-201), WJ-Exo and DMSO. ( B ) Quantitative results for the wound area analysis are presented. (* P < 0.05, ** P < 0.001, *** P < 0.03, **** P < 0.0001 vs. the control group). Data underwent statistical analysis using Two-Way ANOVA followed by Tukey post hoc test for multiple comparisons

Article Snippet: After 48-hour treatment of 4T1 cells (1 × 10 5 cells/96-well plate) with 1 µg of WJ-Exo(S3I-201, 301.4 µM), S3I-201 (337.1 µM), 1 µg of WJ-Exo and DMSO, total RNA, including miRNAs, was extracted with Trizol reagent (Gibco, Germany).

Techniques: Migration, Wound Healing Assay, Control

Journal: Cancer Cell International

Article Title: Enhanced anti-tumor efficacy of S3I-201 in breast cancer mouse model through Wharton jelly- exosome

doi: 10.1186/s12935-024-03501-3

Figure Lengend Snippet: Evaluation of the anti-tumor effects of WJ-Exo, S3I-201 and WJ-Exo(S3I-201) in tumor-bearing mice. ( A ) Determination of tumor volume ( n = 5/group) and ( B ) excision of tumors for weight measurement after 21 days of treatment. ( C ) Monitoring of daily body weight changes in each group. ( D ) Determination of survival rates ( n = 3). The mice received intraperitoneal injections of DMSO, S3I-201 (56 µg/dose), WJ-Exo (10 µg of exosome) and WJ-Exo (S3I-201) (10 µg of exosome loaded with 56 µg/dose S3I-201) on days 10, 12 and 14. Values represent mean ± SD ( n = 8; * P < 0.05, ** P < 0.001, *** P < 0.03, **** P < 0.0001 vs. control). Data underwent statistical analysis using One-Way and Two-Way ANOVA followed by Tukey post hoc test for multiple comparisons, along with Kaplan-Meier estimation for survival analysis

Article Snippet: After 48-hour treatment of 4T1 cells (1 × 10 5 cells/96-well plate) with 1 µg of WJ-Exo(S3I-201, 301.4 µM), S3I-201 (337.1 µM), 1 µg of WJ-Exo and DMSO, total RNA, including miRNAs, was extracted with Trizol reagent (Gibco, Germany).

Techniques: Control

Journal: Cancer Cell International

Article Title: Enhanced anti-tumor efficacy of S3I-201 in breast cancer mouse model through Wharton jelly- exosome

doi: 10.1186/s12935-024-03501-3

Figure Lengend Snippet: Evaluation of splenocyte proliferation and cytokine levels. ( A ) Splenocyte proliferation: Mouse splenocytes (10 6 cells/ml) were stimulated with phytohemagglutinin (PHA) and tumor lysate for 72 h at 37 °C and 5% CO2. Non-stimulated splenocytes under identical conditions were used as controls. Statistical significances indicated with * are compared with the DMSO group. Statistical comparisons between the S3I-201 group and WJ-Exo (S3I-201) were labeled with ####. ( B-E ) Cytokine Production: Cytokine levels in cells cultured with PHA and tumor lysate were measured. Murine splenocytes (10 6 cells/ml) were stimulated for 72 h at 37 °C in 5% CO 2 . Controls comprised non-stimulated splenocytes under the same conditions. Data, representative of at least three ( n = 3) independent experiments per group, were analyzed via Student’s t-test ( n = 3; * p < 0.05, ** p < 0.001, *** p < 0.03, *** p < 0.0001 vs. control). Data underwent statistical analysis using Two-Way ANOVA followed by Tukey post hoc test for multiple comparisons ( A-E )

Article Snippet: After 48-hour treatment of 4T1 cells (1 × 10 5 cells/96-well plate) with 1 µg of WJ-Exo(S3I-201, 301.4 µM), S3I-201 (337.1 µM), 1 µg of WJ-Exo and DMSO, total RNA, including miRNAs, was extracted with Trizol reagent (Gibco, Germany).

Techniques: Labeling, Cell Culture, Control