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Spatial network of major cancer care service flows (volumes ≥ 30 and travel...
Published: 24 May 2022
FIGURE 1 Spatial network of major cancer care service flows (volumes ≥ 30 and travel time ≤ 12 hours) overlaid with 94 CCs. Blue lines represented edges with widths proportional to service flow volumes between ZIP code areas of patients with cancer and ZIP code areas of cancer care providers at th... More
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Delineating the CSA containing six CCs in Los Angeles into sub-CSAs. One in...
Published: 24 May 2022
FIGURE 2 Delineating the CSA containing six CCs in Los Angeles into sub-CSAs. One initial CSA (blue color) in Los Angeles ( A ), three colored sub-CSAs with resolution = 0.3 or 0.4 ( B ), four colored sub-CSAs with resolution = 0.5 ( C ), and five colored sub-CSAs with resolution = 0.6 ( D ). Each... More
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LI of 110 CSAs classified by natural breaks overlaid with 94 CCs. (Nine CSA...
Published: 24 May 2022
FIGURE 3 LI of 110 CSAs classified by natural breaks overlaid with 94 CCs. (Nine CSAs in black boundaries were divided into multiple sub-CSAs in gray boundaries, and the small wad at the east corner of Alaska (inset) was part of the Seattle CSA in the main map. The fragmental white patches represe... More
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Boxplots of nine variables in across two groups: 37 CSAs without CCs and 73...
Published: 24 May 2022
FIGURE 4 Boxplots of nine variables in across two groups: 37 CSAs without CCs and 73 CSAs with CCs. LI ( A ), MSI ( B ), population (in 100,000; C ), population density (persons/km2; D ), urbanization ratio ( E ), average travel time (minutes; F ), area size (in 1,000 km2; G ), area compactness ( H ), and median household income (in $100,000; I ). The Df at the bottom of each boxplot referred to the difference of the mean value of the variable between two groups. *, **, *** significant at 0.05, 0.01, 0.001. (Red dash line and black solid line in the box referred to the mean and median values of each variable across the group.) More
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Correlation coefficient matrices of nine variables across 110 CSAs. Note th...
Published: 24 May 2022
FIGURE 5 Correlation coefficient matrices of nine variables across 110 CSAs. Note that population, area size, and median household income were in logarithmic scales. More
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Supernumerary centrosomes and increased MT growth rates are associated with...
Published: 18 May 2022
FIGURE 1. Supernumerary centrosomes and increased MT growth rates are associated with invasiveness of melanoma cells. A, 3D invasion of melanoma cell lines. Spheroids of the indicated melanoma cell lines were embedded into Matrigel and 3D invasion was followed for 48 hours. Left: Representative images of spheroids and invaded cells. Scale bar, 50 μm. Right: Measurements of the spheroid outgrowth area after 48 hours in Matrigel (mean ± SD, n = 19–23 spheroids, t test). B, Live-cell measurements of MT plus end growth rates in melanoma cells during interphase. Left: Example image of a melanoma cell expressing EB3-GFP and used for measurements of MT growth rates by live-cell microscopy. Right: Determination of MT growth rates in the indicated melanoma cell lines. Scatter dot plots showing average MT growth rates (20 MTs/cell, n = 30 cells, mean ± SD, t test). C, Detection of supernumerary centrosomes in invasive and noninvasive melanoma cells. Top: Representative immunofluorescence microscopy images of melanoma cells with or without supernumerary (>2) centrosomes and analyzed by detecting γ-tubulin as a marker for centrosomes. White triangles mark centrosomes, nuclei were stained by Hoechst. Scale bar, 5 μm. Bottom: Quantification of the proportion of cells with supernumerary centrosomes (mean ± SD, n = 600 cells, t test). D, Representative Western blots detecting PLK4 or STIL overexpression in noninvasive melanoma cells. α-tubulin was detected as a loading control. E, Quantification of the proportion of noninvasive melanoma cells with supernumerary centrosomes upon STIL or PLK4 overexpression. Top: example images of SK-Mel-173 cells with or without STIL or PLK4 overexpression. Centrosomes were stained by anti-γ-tubulin antibodies and nuclei were stained by Hoechst. Scale bar, 5 μm. Bottom: The graph shows the proportion of cells with supernumerary centrosomes (mean ± SD, n = 300 cells, t test). F, Determination of MT plus end growth rates in the indicated noninvasive melanoma cells after PLK4 or STIL overexpression. Scatter dot plots showing average MT growth rates (20 MTs/cell, n = 30 cells, mean ± SD, t test). More
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Supernumerary centrosomes cause increased MT growth rates to promote melano...
Published: 18 May 2022
FIGURE 2. Supernumerary centrosomes cause increased MT growth rates to promote melanoma cell invasion. A, Representative Western blots showing siRNA mediated downregulation of ch-TOG in the indicated invasive melanoma cells. α-tubulin was detected as a protein loading control. B, Determination of MT plus end growth rates in invasive melanoma cell lines after partial siRNA-mediated depletion of ch-TOG. Scatter dot plots show average MT growth rates (20 MTs/cell, mean ± SD, n = 30, t test). C, Measurements of MT growth rates in invasive melanoma cell lines after treatment with DMSO (control) or 0.5 nmol/L Taxol for 16 hours. Noninvasive SK-Mel-173 cells were used as control. Scatter dot plots show average MT growth rates (20 MTs/cell, mean ± SD, n = 30, t test). D, Quantification of the proportion of invasive melanoma cells with supernumerary centrosomes upon partial depletion of ch-TOG or after treatment with Taxol (mean ± SD, n = 300 cells, t-test). E, Invasive 3D outgrowth of spheroids derived from invasive SK-Mel-103 melanoma cells and treated with 0.5 nmol/L Taxol or after siRNA-mediated depletion of ch-TOG. Spheroids were embedded into Matrigel and 3D outgrowth was followed for 48 hours in the presence or absence of treatments. Top: representative microscopy images showing outgrowth of 3D spheroids derived from SK-Mel-103 and SK-Mel-147 cells. Scale bar, 50 μm. Bottom: Quantification of the 3D outgrowth area of spheroids derived from the indicated invasive melanoma cells after treatment with DMSO (control), 0.5 nmol/L Taxol or upon siRNA-mediated knockdown of CKAP5 after 48 hours (mean ± SD, n = 18–26 spheroids, t test). More
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Induction of increased MT growth activity by paracrine signaling.  A,  Mode...
Published: 18 May 2022
FIGURE 3. Induction of increased MT growth activity by paracrine signaling. A, Model depicting the paracrine induction of increased MT growth rates. B, Measurements of MT growth rates in noninvasive SK-Mel-173 cells after cocultivation of noninvasive or invasive melanoma cells for 24 hours. C, Measurements of MT growth rates in the indicated noninvasive melanoma cells after treatment with CM derived from noninvasive or invasive cells for 16 hours. D, Measurements of MT growth rates in the indicated noninvasive melanoma cells after treatment with conditioned media derived from the same cells with or without PLK4 or STIL overexpression. E, Measurements of MT growth rates in noninvasive melanoma cells after treatment with conditioned media derived from the indicated invasive melanoma cells with or without partial depletion of ch-TOG (CKAP5 repression). F, Measurements of MT growth rates in noninvasive SK-Mel-173 cells after treatment with conditioned media derived from the same cells with PLK4 overexpression and concomitant CKAP5 repression. G, Measurements of MT growth rates in noninvasive SK-Mel-173 cells after treatment with conditioned media derived from the same cells with or without CKAP5 overexpression. All scatter dot plots show average MT growth rates (20 MTs/cell, mean ± SD, n = 30, t test). More
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MVs mediate increased MT growth rates.  A,  Measurements of MT growth rates...
Published: 18 May 2022
FIGURE 4. MVs mediate increased MT growth rates. A, Measurements of MT growth rates in noninvasive melanoma cells after treatment with CM derived from noninvasive or invasive cells before and after centrifugation to deplete MVs from the media. B, Measurements of MT growth rates in noninvasive melanoma cells after treatment with conditioned media derived from noninvasive or invasive cells before and after heating the media for 30 minutes at 56°C to denature proteins. All scatter dot plots show average MT growth rates (20 MTs/cell, mean ± SD, n = 30, t test). More
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MVs comprising HER2 mediate enhanced MT growth.  A,  Identification of HER2...
Published: 18 May 2022
FIGURE 5. MVs comprising HER2 mediate enhanced MT growth. A, Identification of HER2/ERBB2 as a constituent of MVs upon induction of supernumerary centrosomes. Isolated MVs derived from noninvasive SK-Mel-173 cells with or without overexpression of PLK4 were subjected to MS analysis. The volcano plot depicts HER2/ERBB2 and HER3/ERBB3 as well as several marker proteins for MVs. B, Detection of HER2 in whole-cell lysates and in MVs derived from noninvasive SK-Mel-173 cells with or without PLK4 overexpression. Representative Western blots detecting HER2, PLK4, GAPDH (loading control for MVs) and α-tubulin (loading control for whole-cell lysates) are shown. C, Representative Western blots showing mild overexpression of HER2 in noninvasive SK-Mel-173 cells. β-actin was detected as a loading control. D, Measurements of MT growth rates in noninvasive melanoma cells after mild overexpression of HER2 in the absence or presence of the irreversible HER2 kinase inhibitor canertinib. Scatter dot plots show average MT growth rates (20 MTs/cell, mean ± SD, n = 30, t test). E, Measurements of MT growth rates in the indicated invasive melanoma cells after treatment with DMSO (control) or the HER2 inhibitor trastuzumab. The scatter dot plots show average MT growth rates (20 MTs/cell, mean ± SD, n = 30, t test). F, 3D outgrowth of spheroids derived from invasive melanoma cells and treated with DMSO (control) or with the HER2 inhibitor canertinib. Left: Representative example images. Scale bar, 25 μm. Right: Quantification of the 3D outgrowth area of spheroids derived from the indicated invasive melanoma cells in the absence or presence of canertinib. The bar graphs show mean values ± SD (n = 19–27 spheroids, t test). G, Measurements of MT growth rates in noninvasive melanoma cells (SK-Mel-173) after treatment with MVs derived from noninvasive (SK-Mel-173) or invasive cells (SK-Mel-147 or SK-Mel-103) that were transiently treated with reversible (trastuzumab) or irreversible (canertinib) HER2 inhibitor. The scatter dot plots show average MT growth rates (20 MTs/cell, mean ± SD, n = 30, t test). More
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