integrinα6  (Santa Cruz Biotechnology)

Journal: Pharmaceuticals

Article Title: Atovaquone Suppresses the Growth of Metastatic Triple-Negative Breast Tumors in Lungs and Brain by Inhibiting Integrin/FAK Signaling Axis

doi: 10.3390/ph14060521

Figure Lengend Snippet: ATQ inhibits integrin-FAK signaling. ( a ) MDA-MB-231, ( b ) HCC1806, ( c ) 4T1 cells were treated with different concentrations of ATQ for 72 h. Representative blots showing concentration-dependent effect of ATQ on integrinα6, integrinβ6, p-Src, Src, p-FAK, FAK, vimentin, MMP-9, Cl caspase 3 and Cl PARP. Actin was used as loading control. Figures shown are the representative blot of at least three independent experiments.

Article Snippet: The antibodies integrinα6 (#3750, rabbit mAb), p-Src (#6943, rabbit mAb), Src (#2108, rabbit mAb), p-FAK (#3283, rabbit mAb), Vimentin (#3932, rabbit mAb), MMP9 (#2270, rabbit mAb), Cl-caspase3 (#9664, rabbit mAb), Cl-PARP (#9541, rabbit mAb) were purchased from Cell Signaling Technologies (Danvers, MA, USA).

Techniques: Concentration Assay, Control

Journal: Pharmaceuticals

Article Title: Atovaquone Suppresses the Growth of Metastatic Triple-Negative Breast Tumors in Lungs and Brain by Inhibiting Integrin/FAK Signaling Axis

doi: 10.3390/ph14060521

Figure Lengend Snippet: Reduction of metastatic breast tumors in lungs by ATQ through inhibition of integrin signaling in intravenous tail vein model. ( a ) Approximately, 0.5 × 10 6 MDA-MB-231-luc cells in 1XPBS were injected intravenously in the tail vein of 4–6 week old athymic nude mice, ( n = 8/per group). Treatment with 50 mg/kg ATQ by oral gavage everyday started at Day 7. Graph showing the luminescence in live mice from control and ATQ treated group. ( b ) Average luminescence of lungs (ex-vivo) isolated from control and ATQ treated mice at the day of termination. Values were plotted as mean ± SEM. ( c ) Representative image of lung from control and ATQ treated mice. The value of luminescence in the control and treated group ranged from 63–877 and 31–288 photons/sec respectively. ( d ) Average weight of mice from control and ATQ treated group throughout the experiment. Tumors were removed after terminating the experiment, homogenized, lysed, and analyzed for integrinα6, integrinβ6, p-Src, vimentin, MMP-9, Cl caspase 3 and Cl PARP by Western blotting. Actin was used as loading control. ( e ) Each lane of blot represents tumor from individual mouse. ( f ) Immunohistochemical staining for integrinβ6 and Cl caspase 3 in tumor lesions in lungs of control and ATQ treated mice. * Statistically significant as compared with control.

Article Snippet: The antibodies integrinα6 (#3750, rabbit mAb), p-Src (#6943, rabbit mAb), Src (#2108, rabbit mAb), p-FAK (#3283, rabbit mAb), Vimentin (#3932, rabbit mAb), MMP9 (#2270, rabbit mAb), Cl-caspase3 (#9664, rabbit mAb), Cl-PARP (#9541, rabbit mAb) were purchased from Cell Signaling Technologies (Danvers, MA, USA).

Techniques: Inhibition, Injection, Control, Ex Vivo, Isolation, Western Blot, Immunohistochemical staining, Staining

Journal: Frontiers in Cell and Developmental Biology

Article Title: Induced Pluripotent Stem Cells Derived From Two Idiopathic Azoospermia Patients Display Compromised Differentiation Potential for Primordial Germ Cell Fate

doi: 10.3389/fcell.2020.00432

Figure Lengend Snippet: Specification of PGCLCs from idiopathic NOA patient-specific iPSCs. (A) Schematic protocol for hPGCLCs specification from idiopathic NOA patient-specific iPSCs and images of hiPSCs, iMeLCs, and floating embryoids containing hPGCLCs. (B) (Top)Fluorescence-activated cell sorting analysis by EpCAM and INTEGRINα6 expression of day 4 embryoids differentiated from NOA iPSCs and normal hiPSCs. P2 and P3 gates (boxed areas) indicate EpCAM/INTEGRINa6-high and -low/no cells, respectively. The percentages of cells in the P2 and P3 gates are shown. (Bottom) Fluorescence-activated cell sorting analysis by c-KIT and CD38 of the two populations on the top classified by EpCAM and INTEGRINa6 expression. (C) Percentage of EpCAM/INTEGRINα6 double-positive cells in days 2, 4, 6, and 8 floating embryoids determined by FACS. Error bars indicate mean ± SD of three independent experiments. (D) Percentage of EpCAM/INTEGRINα6 double-positive cells in day 4 embryoids determined by FACS. The experiments were performed independently for more than six times. Black central line represents the median; boxes represent the 25th and 75th percentiles, and whiskers represent the maximum and minimum. Comparisons were conducted using Wilcoxon signed-ranks test. Asterisk indicates statistically significant differences ( P < 0.05) between the NOA iPSCs and the normal iPSCs.

Article Snippet: The resulting single cells were then incubated with PerCP/Cyanine5.5-conjugated anti-human CD38 (BioLegend, San Diego, CA, United States, cat. no. 356614) and PE/cy7-conjugated anti-human cKIT (BioLegend, cat. no. 313212), or PE-conjugated anti-human EpCAM (eBioscience, Waltham, MA, United States, cat. no. 12-5791-81) and fluorescein isothiocyanate (FITC)–conjugated anti-human INTEGRINα6 (eBioscience, cat. no. 11-0495-82) at 4°C for 30 min. Fluorescence-activated cell sorting analysis was performed using the FACS Calibur system (Becton Dickinson, Franklin Lakes, NJ, United States).

Techniques: Fluorescence, FACS, Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: Induced Pluripotent Stem Cells Derived From Two Idiopathic Azoospermia Patients Display Compromised Differentiation Potential for Primordial Germ Cell Fate

doi: 10.3389/fcell.2020.00432

Figure Lengend Snippet: Differentially expressed gene analysis between PGCLCs derived from NOA and normal hiPSCs. (A) Heat map of gene expression of key PGC-associated genes (early and late stage) and of pluripotency, mesoderm, endoderm, ectoderm, and ICM genes. (B) Numbers of genes upregulated or downregulated at key stages during PGCLC specification of NOA 1106 iPSCs (left) and NOA 1122 iPSCs (right) compared with that of normal iPSCs. (C) Volcano plot of the DEGs in PGCLCs derived from NOA 1106 iPSCs (left) and NOA 1122 iPSCs (right) compared with PGCLCs derived from normal iPSCs. The red dots indicate genes upregulated, and the green dots indicate genes downregulated. (D) Enriched GO terms in the upregulated genes of PGCLCs derived from NOA 1106 iPSCs (left) and NOA 1122 iPSCs (right) compared with PGCLCs derived from normal iPSCs. Primordial germ cell–like cells, the EpCAM, and INTEGRINα6 double-positive cells in day 4 embryoids. (E) Fluorescence-activated cell sorting analysis by annexin V and PI staining for day 4 embryoids differentiated from NOA iPSCs and normal hiPSCs. The percentages of cells in the four quadrants are shown. (F) Apoptotic rates of day 4 embryoids differentiated from NOA iPSCs and normal hiPSCs. Error bars indicate mean ± SD of three independent experiments. Comparisons were conducted using ANOVA. Asterisk indicated statistically significant differences ( P < 0.05) between the NOA iPSCs and the normal iPSCs.

Article Snippet: The resulting single cells were then incubated with PerCP/Cyanine5.5-conjugated anti-human CD38 (BioLegend, San Diego, CA, United States, cat. no. 356614) and PE/cy7-conjugated anti-human cKIT (BioLegend, cat. no. 313212), or PE-conjugated anti-human EpCAM (eBioscience, Waltham, MA, United States, cat. no. 12-5791-81) and fluorescein isothiocyanate (FITC)–conjugated anti-human INTEGRINα6 (eBioscience, cat. no. 11-0495-82) at 4°C for 30 min. Fluorescence-activated cell sorting analysis was performed using the FACS Calibur system (Becton Dickinson, Franklin Lakes, NJ, United States).

Techniques: Derivative Assay, Gene Expression, Fluorescence, FACS, Staining