Sonification of Arabidopsis Mitochondria and Cell Membrane

This movie shows a sonification of a zoomed in section of the <i>Arabidopsis thaliana</i> image volume, with the hyperspectral false color image overlaid on the multichannel image. The spectral differences between the cell membrane and the mitochondria that are obscured by the hyperspectral image are made clear through the sonification

Sonification of Arabidopsis Plastid and Cell Membrane

This movie shows a sonification of the <i>Arabidopsis thaliana</i> image volume, with the hyperspectral false color image overlaid on the multichannel image. The spectral differences between the cell membrane and the plastid that are obscured by the hyperspectral image are made clear through the sonification

Sonification of phantom image volume

In this movie two volumes are shown: on the left is the phantom image volume overlaid with the hyperspectral false color image and on the right is the same phantom volume without the false color overlay. The channels of the right volume are scrolled to visually show the different characteristics of each channel, then the left volume is sonified. While the false color overlay appears homogeneous across the left volume, the sonification reveals the heterogeneous spectral content throughout the image

Sonification of focalcheck beads

This movie shows multichannel image volumes of beads 1,2, and 3 alongside one another, each overlaid with a false color image. While the false color renderings of each volume appear to be the same visually, the sonification of each volume enunciates the subtle spectral difference between beads 1 and 2, and the spectral difference between the core and the edge in bead 3

Video of Calibration

This movie shows a visual comparison of each of the calibration volumes by scrolling through the channels of each volume, then an auditory comparison of each volume for one particular synth

Dense Collagen-I Matrices Enhance Pro-Tumorigenic Estrogen-Prolactin Crosstalk in MCF-7 and T47D Breast Cancer Cells

<div><p>Breast cancers that express estrogen receptor alpha (ERα+) constitute the majority of breast tumors. Estrogen is a major driver of their growth, and targeting ER-mediated signals is a largely successful primary therapeutic strategy. Nonetheless, ERα+ tumors also result in the most breast cancer mortalities. Other factors, including altered characteristics of the extracellular matrix such as density and orientation and consequences for estrogen crosstalk with other hormones such as prolactin (PRL), may contribute to these poor outcomes. Here we employed defined three dimensional low density/compliant and high density/stiff collagen-I matrices to investigate the effects on 17β-estradiol (E2) activity and PRL/E2 interactions in two well-characterized ERα+/PRLR+ luminal breast cancer cell lines <i>in vitro.</i> We demonstrate that matrix density modulated E2-induced transcripts, but did not alter the growth response. However, matrix density was a potent determinant of the behavioral outcomes of PRL/E2 crosstalk. High density/stiff matrices enhanced PRL/E2-induced growth mediated by increased activation of Src family kinases and insensitivity to the estrogen antagonist, 4-hydroxytamoxifen. It also permitted these hormones in combination to drive invasion and modify the alignment of collagen fibers. In contrast, low density/compliant matrices allowed modest if any cooperation between E2 and PRL to growth and did not permit hormone-induced invasion or collagen reorientation. Our studies demonstrate the power of matrix density to determine the outcomes of hormone actions and suggest that stiff matrices are potent collaborators of estrogen and PRL in progression of ERα+ breast cancer. Our evidence for bidirectional interactions between these hormones and the extracellular matrix provides novel insights into the regulation of the microenvironment of ERα+ breast cancer and suggests new therapeutic approaches.</p></div

High density/ stiff collagen matrices decrease E2-induced ERE-luciferase expression in MCF-7 and T47D cells.

<p><b>a)</b> MCF-7 cells stably transfected with an ERE-luciferase construct (MELN) were plated in low density (LD) or high density (HD) collagen and treated ± 1nM E2 for 24h, and luciferase activity determined as described in the Materials and Methods. n = 3, *p<0.05, ***p<0.001. <b>b)</b> T47D cells stably transfected with an ERE-luciferase construct (T47D-KBluc) were treated and harvested as in (a). n = 3, **p<0.01, ***p<0.001. <b>c)</b> Matrix density did not affect ERα expression. Lysates from cells plated in LD or HD collagen were examined for ERα expression. Representative blots shown.</p

Summary of the effects of differences in collagen matrix density on estrogen and PRL crosstalk in breast cancer cells.

<p><b>a)</b> Low density/ compliant collagen matrices permit modest cooperation between E2 and PRL to growth of T47D cells and inhibit E2-induced growth in MCF-7 cells. E2-PRL induced growth is inhibited by 4-OHT, but not SFK inhibitors in both cell lines. Moreover, MCF-7 cells do not invade in response to hormones. These results indicate low density matrices do not allow estrogen and PRL crosstalk to stimulate tumor progression. <b>b)</b> High density/ stiff collagen matrices permit E2-PRL to enhance growth, reduce 4-OHT sensitivity, and drive invasion of MCF7 cells. SFKs are key regulators of the growth response in both cell lines only in stiff collagen matrices. Together, these results indicate that stiff matrices cooperate with E2-PRL crosstalk to fuel processes leading to tumor progression. V = vehicle, ND = not determined; ↑, + indicate relative strength of response.</p

E2+PRL induces invasion only in high density/ stiff collagen matrices.

<p><b>a)</b> MCF-7 cells were cultured in LD or HD collagen on 8µm transwell inserts, treated for 24hr with EtOH vehicle ± 2nM E2, ± 8nM PRL as indicated, and transversed cells were counted. n = 3. Asterisks indicate significant differences between the same treatments in different densities, *p<0.05, ***p<0.001. Different letters represent significant differences (p<0.05) between treatments within each matrix density condition. <b>b)</b> MCF-7 cells were cultured in LD or HD collagen and treated +/- 1nM E2, ± 4nM PRL for 72h. SHG images were collected and analyzed as described in the Materials and Methods. n = 7–8, *p<0.05. c) Representative heat maps of angles of collagen fibers relative to boundaries of cells cultured in low and high matrix densities co-treated with estrogen and PRL. Clear (black) areas represent 0–15 degrees; green, 15–45 degrees; yellow, 45–60 degrees; red, 60–90 degrees.</p

SFKs mediate PRL-augmented growth only in high density/ stiff collagen matrices.

<p><b>a)</b> MCF-7 cells were cultured in LD or HD collagen-1 gels, pretreated for 1h with vehicle or 500nM of the SFK inhibitor, PP1, and then cultured for an additional 72h with the indicated hormones, when cells were counted. n = 3. <b>b)</b> MCF-7 cells were cultured in LD or HD collagen gels, treated for 10min as indicated, and lysates were immunoblotted with the indicated antibodies. Representative blots shown. <b>c)</b> T47D cells were cultured in LD or HD collagen gels, pretreated for 1h with vehicle or 500nM of the SFK inhibitor, PP1, and then cultured for an additional 7d with the indicated hormones, when cells were counted. n = 3. <b>d)</b> T47D cells were cultured in LD or HD collagen gels, treated for 10min as indicated, and lysates were immunoblotted with the indicated antibodies. Representative blots shown. Different letters represent significant differences (p<0.05) within vehicle or PP1 treatments. Asterisks indicate significant differences between the same treatments in different densities, ***p<0.001.</p