Skip to main content
. 2023 Nov 29;14:7759. doi: 10.1038/s41467-023-43519-1

Fig. 3. HDAC8 increases invasion and an amoeboid-like state in melanoma cells.

Fig. 3

a HDAC8 expression increases invasion in a collagen-implanted spheroid assay. EV and HDAC8 expressing melanoma spheroids were plated in collagen for indicated time points. Scale bars = 500 µm. b Spheroid invasion area was quantified using ImageJ software. Significance was determined using a 2-tailed student’s t test with ***p < 0.001. (WM164: p = 0.0002 and SK-MEL-28: p < 0.0001). c HDAC8 expression increases invasion into Matrigel. EV and HDAC8 expressing cells were overlayed onto transwell inserts coated with Matrigel and allowed to invade for 48 h followed by imaging by confocal microscopy. Scale bars = 100 µm. d Invasion area was quantified using ImageJ software. Significance was determined using a 2-tailed student’s t test with **p < 0.01. (p = 0.0028). e HDAC8 increases melanoma survival in shear stress/flow assays. Cells were incubated for 24 h under continual shear stress conditions of 10 dyne/cm2. Cells were stained with Calcein AM/PI and imaged for cell viability. Scale bars = 250 µm. f Data shows numbers of dead cells (PI-positive cells) per image from (e). Significance was determined using a 2-tailed student’s t test with ***p < 0.001. (WM164: p < 0.0001 and SK-MEL-28: p < 0.0001). g HDAC8 leads to the adoption of an amoeboid-like morphology in melanoma cells. HDAC8 and EV expressing cells were plated on collagen overnight followed by brightfield image collection. Scale bars = 250 µm. h HDAC8 enhances transendothelial cell migration. DiI-stained HDAC8 and EV cells were allowed to migrate through a HUVEC coated transwell membrane for 1 h followed by imaging with fluorescence microscopy. Scale bars = 1000 µm. i Quantification of the DiI-stained cells that had migrated through the endothelial cell monolayer (cells per image). Significance was determined using a 2-tailed student’s t test with ***p < 0.001. (WM164: p < 0.0001 and SK-MEL-28: p < 0.0001). Experiments were run 3 independent times with an n of 3 in (d) and an n of 5 in (b), (f), and (i). All data are presented as mean values ± SD. Source data are provided as a Source Data file.