Journal: bioRxiv

Article Title: Microsecond pulse electrical stimulation modulates cell migration

doi: 10.1101/2022.10.23.513372

Figure Lengend Snippet: (a) A graphical illustration showed the effects of μsPEF (pulse width:20 μs, frequency: 10 Hz, duration: 5 s) on the skin wound, promoting cell migration and extracellular matrix remodeling. (b) Representative time-lapse images showing the fibroblasts morphology after μsPEF (i.e., 750 and 1500 V/cm) treatment of different intensity at different time points (i.e., 0, 1 and 2 h). Detached cells are marked by yellow arrow. Scale bar, 50 μm. (c) The line graph representing the cell migration average speed per 1 h from 0 to 12 h after different intensity μsPEF (i.e., 750 and 1500 V/cm) treatment (blue circle: control; orange square: 750 V/cm; pink triangle: 1500 V/cm). Results are presented as mean ± standard deviation with 95% CI (n CTRL =42 cells, n 750 v/cm =48 cells, n 1500 v/cm =47 cells); * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001 versus control by one-way ANOVA for multiple comparisons. (d) Box plot showing the average cell migration speed with different intensity μsPEF (i.e., 750 and 1500 V/cm) and control treatments in 24 h. Results are presented as mean ± standard deviation with 95% CI (n CTRL =470 cells, n 750 V/cm =979 cells, n 1500 V/cm =535 cells). (e) Effects of μsPEF on the secretion of COLA2. The level of collagen type I α2 in cellular supernatants was measured after 48 h. The concentration of COLA2 was 0.109 ± 0.018 ng/mL in the control group, 0.257 ± 0.058 ng/mL in the 750 V/cm group and 0.363 ± 0.034 ng/mL in the 1500 V/cm group. Results are presented as mean ± standard deviation with 95% CI (n=5); * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001 versus control by one-way ANOVA for multiple comparisons. (f) Effects of μsPEF on the expression of FGF2. The level of FGF2 in cellular supernatants was measured after 48 h. The concentration of FGF2 was 44.139 ± 0.360 ng/mL in the control group, 48.012 ± 1.488 ng/mL in the 750 V/cm group and 48.523 ± 1.944 ng/mL in the 1500 V/cm group. Results are presented as mean ± standard deviation with 95% CI (n=3); ns=0.7329, * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001 versus control by one-way ANOVA for multiple comparisons.

Article Snippet: Then, the cells were cultured in serum-free medium for 48 h. The content of type I α collagen and basic fibroblast growth factor (FGF-2) in the supernatant were measured using commercially available Human COL1A2 (Collagen Type I Alpha 2) ELISA Kit (Elabscience, Wuhan, China) and Human bFGF/FGF2 (Basic Fibroblast Growth Factor) ELISA Kit (Elabscience, Wuhan, China) according to the manufacturer’s protocol respectively.

Techniques: Migration, Control, Standard Deviation, Concentration Assay, Expressing

Journal: Breast Cancer Research : BCR

Article Title: STAT5 is activated in macrophages by breast cancer cell-derived factors and regulates macrophage function in the tumor microenvironment

doi: 10.1186/s13058-021-01481-0

Figure Lengend Snippet: Tumor cell-derived GM-CSF activates STAT5 in macrophages. A qRT-PCR analysis for GM-CSF in TNBC (Hs578T and MDA-MB-231), ER + (MCF7) and HER2 + (BT-474) human breast cancer cells relative to expression in MCF-10A cells. B ELISA analysis for GM-CSF in CM collected from MCF-10A, MDA-MB-231, Hs578T, MCF7, and BT-474 cells. C ELISA analysis for GM-CSF in CM collected from 4T1 cells and B/B-stimulated HC11, HC11/R1, and HC11/R1-LM cells relative to EtOH controls. D Immunoblot analysis for pSTAT5, TSTAT5, and β-tubulin in BMDMs treated with No CM, tumor CM (4T1 or HC11/R1 BB), or tumor CM incubated for 1 h at 37 °C with 2.5 µg/mL neutralizing GM-CSF antibody (⍺GM-CSF Ab)

Article Snippet: STAT5 fl/fl DCM and STAT5 cKO DCM was subjected to mouse Type I Collagen ELISA (Novus Biologicals).

Techniques: Derivative Assay, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, Western Blot, Incubation

Journal: Breast Cancer Research : BCR

Article Title: STAT5 is activated in macrophages by breast cancer cell-derived factors and regulates macrophage function in the tumor microenvironment

doi: 10.1186/s13058-021-01481-0

Figure Lengend Snippet: STAT5 deletion in macrophages enhances tumor-promoting phenotype and impacts tumor cell migration and metastasis. A qRT-PCR analysis of genes of interest from RNA-seq associated with tumor-promoting pathways in rmGM-CSF or 4T1 CM-treated STAT5 fl/fl (blue) and STAT5 cKO (red) BMDMs. Unpaired t-test was used for statistical analysis. B Mouse Type 1 Collagen ELISA in STAT5 fl/fl unstimulated or 4T1 CM-stimulated STAT5 fl/fl and STAT5 cKO macrophage double CM (DCM). Data were analyzed using one-way ANOVA and Tukey’s multiple comparison test. C Representative immunoblot of pFAK, total FAK (FAK), and β-tubulin in 4T1 cells cultured alone or co-cultured with STAT5 fl/fl or STAT5 cKO BMDMs for 4 h. Densitometry analysis relative to loading control. D Migration analysis of 4T1 cells cultured alone or co-cultured with STAT5 fl/fl or STAT5 cKO BMDMs after 20 h. Cell counts relative to 4T1 alone in triplicate. E Kaplan–Meier curves of 4T1 cells co-injected with either STAT5 fl/fl (n = 8) or STAT5 cKO (n = 7) BMDMs in WT BALB/c mice. % Survival on Y-axes indicates proportion of mice reaching tumor size endpoint of 1cm 3 . F Quantified metastasis in H&E-stained lung sections. Lungs were sectioned at 3 different depths per mouse and analyzed for percent metastatic area per tissue section. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Scale bar: 50 μm

Article Snippet: STAT5 fl/fl DCM and STAT5 cKO DCM was subjected to mouse Type I Collagen ELISA (Novus Biologicals).

Techniques: Migration, Quantitative RT-PCR, RNA Sequencing, Enzyme-linked Immunosorbent Assay, Comparison, Western Blot, Cell Culture, Control, Injection, Staining