Journal: BMC Cancer

Article Title: Fascin-1 enhances experimental osteosarcoma tumor formation and metastasis and is related to poor patient outcome

doi: 10.1186/s12885-019-5303-3

Figure Lengend Snippet: Validation and characterization of OS cell lines with altered Fascin-1 expression. ( a ) Western blot analysis with antibodies to Fascin-1 (top panel), to V5 (middle panel), and to GAPDH as a loading control (lower panel) of protein extracts from SaOS-2 (left panel) and 143B (right panel) cells stably transduced with a scrambled control ShRNA (Ctrl ShRNA), a Fascin-1-specific ShRNA (ShFascin-1), a pLenti6/V5-DEST empty vector (EV), or pLenti6/V5-DEST-Fascin-1 (Fascin-1). ( b ) SaOS-2/WT (upper row), SaOS-2/Fascin-1 (middle row), and SaOS-2/ShFascin-1 (bottom row) cells stained with anti-Fascin-1 (green), with Alexa-633-phalloidin (filamentous actin, red), and with NucBlue (nuclei in blue). ( c ) While silencing Fascin-1 reduces the perimeter of the cells in both cases, overexpression has little impact ( n = 34–57)

Article Snippet: Endogenous and V5-tagged Fascin-1 protein and GAPDH were detected using mouse monoclonal Fascin-1 (1:1000; DAKO), V5 antibody (1:5000; Invitrogen), and rabbit polyclonal anti-GAPDH antibody (1:5000; Santa Cruz Biotechnologies) and corresponding horseradish peroxidase-conjugated secondary antibodies (Santa Cruz Biotechnologies).

Techniques: Expressing, Western Blot, Stable Transfection, Transduction, shRNA, Plasmid Preparation, Staining, Over Expression

Journal: Development (Cambridge, England)

Article Title: Fascin 1 is transiently expressed in mouse melanoblasts during development and promotes migration and proliferation

doi: 10.1242/dev.089789

Figure Lengend Snippet: Fascin 1 overexpression increases melanocyte lamellipodium dynamics. ( A ) GFP fluorescence and transmitted light photo of M4F melanocytes indicate that fascin 1-enriched filopodia (white arrows) are interspersed in the lamellipodium. Scale bar: 5 μm. ( B ) Cell migration speed generated from three independent experiments. Data are generated from 98 M4 cells and 135 M4F cells. ( C ) Representative kymograph pictures of M4 (top) and M4F (bottom) showing M4F cells have higher rates of lamellipodia protrusions. Lamellipodial kymographs are analysed by drawing a one pixel wide line (white line in top right box) at a 90° angle across a lamellipodium in the direction of protrusion. Pixel densities along this pixel-wide line are recorded at 1-second intervals for a period of 300 seconds, resulting a time-space image. Scale bars for time and space are shown in the top image. Within the kymograph, a steeper slope of a protrusion corresponds to higher rates for lamellipodia protrusion. Distance extended (d) was measured against lamellipodial persistence (p), defined as the amount of time that the cell spends persistently extending a lamellipod before pausing or entering a retraction phase. Speed of protrusion is shown as a broken line and represents the distance protruded (d) divided by the time of protrusion (p). Frequency is measured by counting the number of lamellipodia per minute. ( D ) Frequency of lamellipodial protrusion events in M4 and M4F cells. ( E-G ) Speed (E), persistence (F) and distance (G) of individual lamellipodial protrusions. Data for M4 cells are from 273 protrusion events and for M4F cells from 144 protrusion events. In the box and whisker plots, horizontal line indicates median, box indicates interquartile range and whiskers indicate maximum value to minimum value. Mann-Whitney test; *** P <0.001; * P <0.05; n.s., not significant.

Article Snippet: The following primary antibodies were used: goat anti-TRP2 antibody (DCT) (D-18, 1:200, Santa Cruz) combined with rabbit anti-cleaved caspase 3 (Asp175) (5A1E, 1:100, NEB) or rabbit anti-phospho-histone H3 (ser10) (1:100, Cell Signaling), mouse anti-BrdU (5-bromo-2′-deoxyuridine) antibody (1:100, DAKO), rabbit anti-cyclin D1 antibody (EP12, 1:100, DAKO) and rabbit anti-fascin 1 (1:100, Sigma-Aldrich).

Techniques: Over Expression, Fluorescence, Migration, Generated, Whisker Assay, MANN-WHITNEY

Journal: Development (Cambridge, England)

Article Title: Fascin 1 is transiently expressed in mouse melanoblasts during development and promotes migration and proliferation

doi: 10.1242/dev.089789

Figure Lengend Snippet: Fascin 1 loss causes hypopigmentation. ( A ) Fascin1 expression level in skin lysates from fascin 1 wild-type, fascin 1 -/+ and fascin 1 -/- embryos (E14.5). ( B ) White patch phenotype of fascin 1 -/- mice in C57BL/6 background. Small white patches are defined as a few white hairs to a very thin white straight line in the middle of the abdomen. Big white patches are defined as diversely shaped patches normally covering 5-20% of the abdomen. ( C ) DCT and fascin 1 immunofluorescence pictures of transversely sectioned embryos. Broken white lines indicate the epidermal (Ep)-dermal (De) border (green: DCT, red: fascin1, blue: DAPI); yellow arrowhead indicates a melanoblast with a protrusion and white arrowhead indicates a basal keratinocyte. Scale bars: 10 μm. ( D , E ) Immunohistochemistry using DCT antibody and haemoxylin counterstains of wild type (D) and fascin 1 -/- (E) dorsal and ventral skin sections. HF, hair follicles. Scale bars: 100 μm. ( F ) Tails and toes of wild-type and fascin 1 -/- C57BL/6 mice.

Article Snippet: The following primary antibodies were used: goat anti-TRP2 antibody (DCT) (D-18, 1:200, Santa Cruz) combined with rabbit anti-cleaved caspase 3 (Asp175) (5A1E, 1:100, NEB) or rabbit anti-phospho-histone H3 (ser10) (1:100, Cell Signaling), mouse anti-BrdU (5-bromo-2′-deoxyuridine) antibody (1:100, DAKO), rabbit anti-cyclin D1 antibody (EP12, 1:100, DAKO) and rabbit anti-fascin 1 (1:100, Sigma-Aldrich).

Techniques: Expressing, Immunofluorescence, Immunohistochemistry

Journal: Development (Cambridge, England)

Article Title: Fascin 1 is transiently expressed in mouse melanoblasts during development and promotes migration and proliferation

doi: 10.1242/dev.089789

Figure Lengend Snippet: Loss of fascin 1 results in fewer melanoblasts by E13.5 and E15.5. ( A ) Wild-type and fascin 1 -/- embryo pictures and cropped areas used for quantitation (E11.5). ( B ) Cell number in dorsal-ventral part of E11.5 embryos (wild type, n =8; -/+, n =6; -/-, n =3). Kruskal-Wallis one-way ANOVA test and Mann-Whitney test, n.s., not significant. ( C ) Wild-type and fascin 1 -/- pictures of whole embryos (E15.5) with cropped, gridded subareas for quantitation. ( D , E ) Melanoblast number in dorsal-ventral regions of E13.5 (D) and E15.5 (E) embryos. ( F ) Wild-type and fascin 1 -/- pictures of the forelimbs with cropped, gridded subareas for quantitation. ( G , H ) Melanoblast number in the forelimbs of E13.5 (G) and E15.5 (H) embryos. Results are expressed as means±s.e.m. (four to six embryos for each genotype), Kruskal-Wallis one-way ANOVA tests were used to compare melanoblast numbers between wild type, -/+ and -/- in each subarea and Mann-Whitney tests were used to compare between two groups, * P <0.05; ** P <0.01; n.s., not significant.

Article Snippet: The following primary antibodies were used: goat anti-TRP2 antibody (DCT) (D-18, 1:200, Santa Cruz) combined with rabbit anti-cleaved caspase 3 (Asp175) (5A1E, 1:100, NEB) or rabbit anti-phospho-histone H3 (ser10) (1:100, Cell Signaling), mouse anti-BrdU (5-bromo-2′-deoxyuridine) antibody (1:100, DAKO), rabbit anti-cyclin D1 antibody (EP12, 1:100, DAKO) and rabbit anti-fascin 1 (1:100, Sigma-Aldrich).

Techniques: Quantitation Assay, MANN-WHITNEY

Journal: Development (Cambridge, England)

Article Title: Fascin 1 is transiently expressed in mouse melanoblasts during development and promotes migration and proliferation

doi: 10.1242/dev.089789

Figure Lengend Snippet: Fascin 1 loss delays melanoblast and melanoma cell cycle progression. ( A ) Percentage of BrdU-positive DCT labelling melanoblasts (three embryos for each genotype, unpaired Student’s t -test, * P <0.05). ( B ) Percentage of phospho-histone H3 (ser10) (pH3)-positive DCT labelling melanoblasts (five embryos for each genotype, unpaired Student’s t -test, ** P <0.01). ( C ) Percentage of nuclear cyclin D1-positive DCT labelling melanoblasts (wild-type, n =7; -/-, n =4; unpaired Student’s t -test, * P <0.05). Fifty to 100 cells are counted per embryo in A-C. ( D ) Cell cycle distribution of MV3 melanoma cells as measured with BrdU/PI FACS analysis. Results are from four independent experiments (mean±s.e.m., unpaired Student’s t -test, ** P <0.01). ( E ) Percentage of BrdU-positive MV3 cells at the indicated time points of nocodazole trapping showing fewer proliferating sifascin1 knockdown cells. Results are from three to six independent experiments (means±s.e.m., unpaired Student’s t -test, * P <0.05). The sifascin1 line is a pool of two different siRNAs that target fascin 1.

Article Snippet: The following primary antibodies were used: goat anti-TRP2 antibody (DCT) (D-18, 1:200, Santa Cruz) combined with rabbit anti-cleaved caspase 3 (Asp175) (5A1E, 1:100, NEB) or rabbit anti-phospho-histone H3 (ser10) (1:100, Cell Signaling), mouse anti-BrdU (5-bromo-2′-deoxyuridine) antibody (1:100, DAKO), rabbit anti-cyclin D1 antibody (EP12, 1:100, DAKO) and rabbit anti-fascin 1 (1:100, Sigma-Aldrich).

Techniques:

Journal: Development (Cambridge, England)

Article Title: Fascin 1 is transiently expressed in mouse melanoblasts during development and promotes migration and proliferation

doi: 10.1242/dev.089789

Figure Lengend Snippet: Fascin 1 loss reduces melanoblast cell migration speed and pseudopod generation (E14.5). ( A ) Picture series of melanoblast migration (wild type and fascin 1 -/- ). Red arrows indicate the protruding pseudopods. Scale bars: 10 μm. ( B ) Cell tracks of wild type and fascin 1 -/- representative movies. Red lines represent cells with speed slower than 0.5 μm/minute. ( C ) Relative cell area expressed as pixels and measured with ImageJ line tool. Between 170 and 200 cells are analysed. ( D , E ) Cell speed (D) and frequency distribution of cell speed (E) of wild-type and fascin 1 -/- melanoblasts, as measured with ImageJ plug-in chemotaxis tool. ( F ) Euclidean migration distance in wild-type and fascin 1 -/- melanoblasts. Data are generated from three movies (three different embryos) and 230-250 cells from each genotype are analysed in total. ( G ) Detailed pictures with cell protrusions. White arrowheads indicate the midpoint of the pseudopods where the diameter was measured. ( H , I ) Protrusion numbers that melanoblasts generate in 4 hours (H) and pseudopod diameter (I) in still z stacked pictures. Pseudopods of 80-130 cells from three embryos were analysed. Scale bars: 10 μm. In the box and whisker plots, horizontal line indicates median, box indicates interquartile range and whiskers indicate maximum value to minimum value. Mann-Whitney test; ** P <0.01; *** P <0.001.

Article Snippet: The following primary antibodies were used: goat anti-TRP2 antibody (DCT) (D-18, 1:200, Santa Cruz) combined with rabbit anti-cleaved caspase 3 (Asp175) (5A1E, 1:100, NEB) or rabbit anti-phospho-histone H3 (ser10) (1:100, Cell Signaling), mouse anti-BrdU (5-bromo-2′-deoxyuridine) antibody (1:100, DAKO), rabbit anti-cyclin D1 antibody (EP12, 1:100, DAKO) and rabbit anti-fascin 1 (1:100, Sigma-Aldrich).

Techniques: Migration, Chemotaxis Assay, Generated, Whisker Assay, MANN-WHITNEY

Journal: Molecular Biology of the Cell

Article Title: Survey of cancer cell anatomy in nonadhesive confinement reveals a role for filamin-A and fascin-1 in leader bleb–based migration

doi: 10.1091/mbc.E21-04-0174

Figure Lengend Snippet: Analysis of the effects of the overexpression of actin regulatory proteins on leader bleb formation and cell migration. A375M melanoma cells cultured on BSA (1 µg/ml)-treated coverslips with confinement by a PDMS pad resting on 3 µm beads to define the confinement height and induce leader bleb–based motility. Cells were transiently coexpressing various mEmerald (mEm)-tagged actin regulatory proteins or GFP control together with FusionRed-F-Tractin. Analysis (A–I) presented for cells in which the nucleus was localized in the bleb; see Supplemental Figure 5 for analysis of cells with the nucleus in the cell body. (A–I) Fusion proteins expressed are color coded: GFP control (white); actin assembly-regulating proteins (red, mEmerald-ARP3, mEmerald-cortactin, mEmerald-mDia1, mEmerald-VASP); actomyosin contraction-regulating proteins (green, mEmerald-non-muscle myosin IIA, mEmerald non-muscle-myosin IIB, mEmerald-tropomyosin 2, mEmerald-tropomyosin 4); actin cross-linking proteins (blue, mEmerald-α2-spectrin, mEmerald-α-actinin-1, mEmerald-fascin-1, mEmerald-filamin-A. Average length (A) and width (B) of leader bleb and cell motility speed (E); n (cells) is shown below categories. Error is SEM. Statistical significance was determined by one-way ANOVA. ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001, NS not significant. (C) Rose plots of representative migration tracks (8 h, 10 min intervals) of cells overexpressing the noted fusion protein and undergoing LBBM; the number of cells is noted on each plot. (D) Fraction of cells undergoing leader bleb–based motility with the nucleus localized in the cell body vs. the bleb. The number of cells analyzed ( n ) is shown below each category. Average mean squared displacement over time for cells undergoing LBBM and overexpressing (F) aII spectrin (G) a-actinin (H) fascin-1 or (I) filamin-A; n (cells) = 29 (a-2-spectrin), 42 (α-actinin-1, 34 (fascin-1), and 37 (filamin-A) cells.

Article Snippet: The following antibodies were used: rabbit anti–filamin-A (1:1000; Sigma-Aldrich; HPA002925), rabbit anti–fascin-1 (1:1000; Sigma-Aldrich; HPA005723), rabbit anti-paxillin (1:1000; Invitrogen; 10029-1-IG), rabbit anti-cortactin (1:1000; Millipore-Sigma; SAB4500766), mouse tropomyosin (1:1000; Millipore-Sigma; T2780), rabbit anti-GAPDH (1:2000; Clone 14C10; Cell Signaling; 2118S).

Techniques: Over Expression, Migration, Cell Culture

Journal: Molecular Biology of the Cell

Article Title: Survey of cancer cell anatomy in nonadhesive confinement reveals a role for filamin-A and fascin-1 in leader bleb–based migration

doi: 10.1091/mbc.E21-04-0174

Figure Lengend Snippet: Effects of filamin-A and fascin-1 knockdown on cell morphometrics, mechanics, and migration in nonadhesive confinement. (A–L) A375M cells were treated with nontargeting siRNAs (siCtrl) or siRNA targeting human fascin-1 (si fascin-1) or filamin-A (si filamin-A) with or without the additional expression of mEmerald-fascin-1 (si fascin-1+rescue) or filamin-A (si fascin-1+rescue). (A) Western blot analysis of cell lysates. Blots were probed with antibodies specific to fascin-1, filamin-A, GFP, or GAPDH. (B) Quantitative analysis of relative protein level from Western blots normalized to GAPDH and compared with siControl, measured from three independent experiments. (C) Immunofluorescence of endogenous fascin-1 (top) or filamin-A (bottom) (green) together with phalloidin staining of F-actin (red) and DAPI staining of DNA (blue) in cells under nonadhesive confinement. Scale bars = 10 µm. (D) Percentage of cells exhibiting leader bleb morphology ( n = minimum of 50 cells per condition from N = 3 experiments). (E, F, J–L) A375M melanoma cells cultured on BSA (1 µg/ml)-treated coverslips with confinement by a PDMS pad resting on 3 mm beads to define the confinement height and induce leader bleb–based motility. (E, F) Average length (E) and width (F) of leader bleb. (G–I). Schematic representation of the AFM-based assay used for determining cell cortex tension and intracellular pressure in A375M cells plated on nonadhesive or uncoated glass. k c , cantilever spring constant; d , cantilever deflection; Z , piezo Z displacement (G). Tukey box plots of cortex tension (H) and intracellular hydrostatic pressure (I), where the + and line denote the mean and median, respectively. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001, NS not significant p ≤ 0.05. (J). Rose plots of representative migration tracks (4 h, 10 min intervals) of cells undergoing LBBM; the number of cells is noted on each plot. (K) Average speed of cells undergoing LBBM. (L) Average mean squared displacement over time; n (cells) = 58 (siControl [nontargeting], 76 (sifascin-1), 38 (sifascin1+Rescue), and 51 (sifilamin-A) and 51 (sifilamin-A+Rescue). All data are representatives of at least three independent experiments. Error is SEM. The statistical significance was determined by two-tailed Student’s t tests, one-way ANOVA, and/or Mann–Whitney. ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001, NS not significant.

Article Snippet: The following antibodies were used: rabbit anti–filamin-A (1:1000; Sigma-Aldrich; HPA002925), rabbit anti–fascin-1 (1:1000; Sigma-Aldrich; HPA005723), rabbit anti-paxillin (1:1000; Invitrogen; 10029-1-IG), rabbit anti-cortactin (1:1000; Millipore-Sigma; SAB4500766), mouse tropomyosin (1:1000; Millipore-Sigma; T2780), rabbit anti-GAPDH (1:2000; Clone 14C10; Cell Signaling; 2118S).

Techniques: Migration, Expressing, Western Blot, Immunofluorescence, Staining, Cell Culture, Two Tailed Test, MANN-WHITNEY

Journal: Molecular Biology of the Cell

Article Title: Survey of cancer cell anatomy in nonadhesive confinement reveals a role for filamin-A and fascin-1 in leader bleb–based migration

doi: 10.1091/mbc.E21-04-0174

Figure Lengend Snippet: Role of filamin-A and fascin-1 in leader bleb formation and migration in nonadhesive confinement. Model for actin filament cross-linking proteins; fascin-1 and filamin-A effects on leader bleb formation, mechanics and LBBM. Leader bleb cell showing reduction and rescue of fascin-1 (top arrows) and the effects on cell morphology, cortical tension, intracellular pressure, and cell migration speed and directionality in melanoma A375M cells. Excess fascin-1 shows no change in bleb size, while the reduction of fascin-1 results in an increase in leader bleb size and decreased motility. Leader bleb cell showing reduction and rescue of filamin-A (bottom arrows) and the effects on cell morphology, cortical tension, intracellular pressure, and cell migration speed and directionality in melanoma A375M2 cells. Excess filamin-A results in an increase in bleb size due to its linkage to cortex and contractility of actomyosin. Reduction of filamin-A shows an increase in overall blebs but no production of leader bleb and decrease in cell speed and directionality with partial rescue, due to lack of pressure production.

Article Snippet: The following antibodies were used: rabbit anti–filamin-A (1:1000; Sigma-Aldrich; HPA002925), rabbit anti–fascin-1 (1:1000; Sigma-Aldrich; HPA005723), rabbit anti-paxillin (1:1000; Invitrogen; 10029-1-IG), rabbit anti-cortactin (1:1000; Millipore-Sigma; SAB4500766), mouse tropomyosin (1:1000; Millipore-Sigma; T2780), rabbit anti-GAPDH (1:2000; Clone 14C10; Cell Signaling; 2118S).

Techniques: Migration

Journal: The Journal of Biological Chemistry

Article Title: Transducer of Cdc42-dependent Actin Assembly Promotes Epidermal Growth Factor-induced Cell Motility and Invasiveness *

doi: 10.1074/jbc.M110.157974

Figure Lengend Snippet: Toca-1 knockdown cells have defects in EGF-induced filopodia formation. A, representative epifluorescence micrographs are shown for A431 vector control (a and c) and Toca-1 knockdown cells (shRNA2; b and d) plated on gelatin-coated coverslips prior to serum starvation (a and b) and treatment with EGF (100 ng/ml for 15 min; c and d). Cells were fixed in methanol and permeabilized, and filopodia were stained using fascin mAb. B, the graph depicts the average number of filopodia per cell ± S.E. (error bars) between cell lines in each condition (n = 20–30 cells). *, statistically significant differences (p < 0.05) between Toca-1 knockdown and vector control cells based on paired Student's t test. C, the graph depicts the average length of filopodia ± S.E. between cell lines in each condition (n = 20 cells for −EGF, and n = 49 for +EGF). *, statistically significant difference (p < 0.05) between Toca-1 knockdown and vector control cells based on paired Student's t test (n = 40 cells).

Article Snippet: Fascin staining was performed with mouse anti-fascin (1:50; Chemicon) on methanol-fixed cells.

Techniques: Plasmid Preparation, Staining

Journal: BMC Cancer

Article Title: Fascin-1 enhances experimental osteosarcoma tumor formation and metastasis and is related to poor patient outcome

doi: 10.1186/s12885-019-5303-3

Figure Lengend Snippet: Kaplan-Meier analysis correlating immunohistochemical staining of Fascin-1 in human OS tissues with overall survival of the patients. ( A ) Representative images of TMA sections showing entire spots (upper panel) and higher magnification (lower panel) with non-detectable ( a ), weak ( b ), moderate ( c ), and intense ( d ) Fascin-1 immunostaining. ( B ) Overall survival of OS patients with non-detectable (Fascin-1 neg) or detectable (Fascin-1 pos) immunostaining of tumor tissues. ( C ) Overall survival of patients without (Mets neg) or with (Mets pos) metastases and Fascin-1 neg or Fascin-1 pos tumors

Article Snippet: Endogenous and V5-tagged Fascin-1 protein and GAPDH were detected using mouse monoclonal Fascin-1 (1:1000; DAKO), V5 antibody (1:5000; Invitrogen), and rabbit polyclonal anti-GAPDH antibody (1:5000; Santa Cruz Biotechnologies) and corresponding horseradish peroxidase-conjugated secondary antibodies (Santa Cruz Biotechnologies).

Techniques: Immunohistochemical staining, Staining, Immunostaining

Journal: BMC Cancer

Article Title: Fascin-1 enhances experimental osteosarcoma tumor formation and metastasis and is related to poor patient outcome

doi: 10.1186/s12885-019-5303-3

Figure Lengend Snippet: Validation and characterization of OS cell lines with altered Fascin-1 expression. ( a ) Western blot analysis with antibodies to Fascin-1 (top panel), to V5 (middle panel), and to GAPDH as a loading control (lower panel) of protein extracts from SaOS-2 (left panel) and 143B (right panel) cells stably transduced with a scrambled control ShRNA (Ctrl ShRNA), a Fascin-1-specific ShRNA (ShFascin-1), a pLenti6/V5-DEST empty vector (EV), or pLenti6/V5-DEST-Fascin-1 (Fascin-1). ( b ) SaOS-2/WT (upper row), SaOS-2/Fascin-1 (middle row), and SaOS-2/ShFascin-1 (bottom row) cells stained with anti-Fascin-1 (green), with Alexa-633-phalloidin (filamentous actin, red), and with NucBlue (nuclei in blue). ( c ) While silencing Fascin-1 reduces the perimeter of the cells in both cases, overexpression has little impact ( n = 34–57)

Article Snippet: Endogenous and V5-tagged Fascin-1 protein and GAPDH were detected using mouse monoclonal Fascin-1 (1:1000; DAKO), V5 antibody (1:5000; Invitrogen), and rabbit polyclonal anti-GAPDH antibody (1:5000; Santa Cruz Biotechnologies) and corresponding horseradish peroxidase-conjugated secondary antibodies (Santa Cruz Biotechnologies).

Techniques: Expressing, Western Blot, Stable Transfection, Transduction, shRNA, Plasmid Preparation, Staining, Over Expression

Journal: BMC Cancer

Article Title: Fascin-1 enhances experimental osteosarcoma tumor formation and metastasis and is related to poor patient outcome

doi: 10.1186/s12885-019-5303-3

Figure Lengend Snippet: Migration assessed by wound healing . Overexpression of Fascin-1 in SaOS-2 ( a ) or in 143B (b) OS cells increases significantly the migration ability. Similarly, silencing of Fascin-1 slightly decreased the migratory rate of SaOS-2 ( a ) and 143B ( b ) cells. Results are the mean ± SEM of at least three independent experiments. ( c ) Zymography analysis showing increased MMP-9 activity in SaOS-2/Fascin-1 (left panel) and in 143B/Fascin-1 (right panel) in comparison to control SaOS-1/EV and 143B/EV cells respectively. Results are the mean ± SEM of three independent experiments

Article Snippet: Endogenous and V5-tagged Fascin-1 protein and GAPDH were detected using mouse monoclonal Fascin-1 (1:1000; DAKO), V5 antibody (1:5000; Invitrogen), and rabbit polyclonal anti-GAPDH antibody (1:5000; Santa Cruz Biotechnologies) and corresponding horseradish peroxidase-conjugated secondary antibodies (Santa Cruz Biotechnologies).

Techniques: Migration, Over Expression, Zymography, Activity Assay

Journal: BMC Cancer

Article Title: Fascin-1 enhances experimental osteosarcoma tumor formation and metastasis and is related to poor patient outcome

doi: 10.1186/s12885-019-5303-3

Figure Lengend Snippet: Fascin-1 overexpression in SaOS-2 cells promotes intratibial primary tumor growth and lung metastasis in SCID mice. ( a ) Representative X-ray images of tumor-bearing hind limbs of mice injected with SaOS-2/EV cells (upper panel) or with SaOS-2/Fascin-1 (lower panel). The images show primary tumor appearance on indicated days after tumor cell injection. ( b ) Representative X-ray images of tumor-bearing hind limbs of mice injected with SaOS-2/Ctrl ShRNA cells (upper panel) or with SaOS-2/ShFascin-1 cells (lower panel). ( c ) Mean primary tumor growth over time in mice intratibially injected with SaOS-2/EV cells (black), with SaOS-2/Fascin-1 cells (red), with SaOS-2/Ctrl ShRNA (grey) or with SaOS-2/ShFascin-1 blue). ( d ) Mean number ± SEM of metastatic lesions in the lungs

Article Snippet: Endogenous and V5-tagged Fascin-1 protein and GAPDH were detected using mouse monoclonal Fascin-1 (1:1000; DAKO), V5 antibody (1:5000; Invitrogen), and rabbit polyclonal anti-GAPDH antibody (1:5000; Santa Cruz Biotechnologies) and corresponding horseradish peroxidase-conjugated secondary antibodies (Santa Cruz Biotechnologies).

Techniques: Over Expression, Injection, shRNA

Journal: BMC Cancer

Article Title: Fascin-1 enhances experimental osteosarcoma tumor formation and metastasis and is related to poor patient outcome

doi: 10.1186/s12885-019-5303-3

Figure Lengend Snippet: Fascin-1 overexpression in 143B cells promotes intratibial primary tumor growth and lung metastasis in SCID mice. (a) Representative X-ray images of tumor-bearing hind limbs of mice injected with 143B/EV cells (upper panel) or with 143B/Fascin-1 (lower panel). The images show primary tumor appearance on indicated days after tumor cell injection. (b) Representative X-ray images of tumor-bearing hind limbs of mice injected with 143B/Ctrl ShRNA cells (upper panel) or with 143B/ShFascin-1 cells (lower panel). (c) Mean primary tumor growth over time in mice intratibially injected with 143B/EV cells (black), with 143B/Fascin-1 cells (red), with 143B/Ctrl ShRNA (grey) or with 143B/ShFascin-1 blue). (d) Quantification of the number of metastatic lesions in lungs

Article Snippet: Endogenous and V5-tagged Fascin-1 protein and GAPDH were detected using mouse monoclonal Fascin-1 (1:1000; DAKO), V5 antibody (1:5000; Invitrogen), and rabbit polyclonal anti-GAPDH antibody (1:5000; Santa Cruz Biotechnologies) and corresponding horseradish peroxidase-conjugated secondary antibodies (Santa Cruz Biotechnologies).

Techniques: Over Expression, Injection, shRNA