Journal: bioRxiv

Article Title: YAP localization mediates mechanical adaptation of human cancer cells during extravasation in vivo

doi: 10.1101/2023.11.14.567015

Figure Lengend Snippet: (a) Scheme of in vivo cancer cell mechanical measurement at zebrafish brain by optical trap (OT) where cancer cells with internalized 1 µm beads are injected into the circulation of zebrafish at 2 days-post-fertilization (2dpf) for mechanical mapping at 3dpf and 1 day-post-injection (dpi). (b) Images of capillary arrested (intravascular) and extravasated cancers (blue) inside of zebrafish, Tg(flk:mCherry/MRC1a:EGFP), brain at 3dpf/1dpi along with 1 day-post-extravasated (dpe) from 4dpf/2dpi. (c) Tracking of migration of human breast cancer cell (MDA-MB-231) in extravasation mimicking microfluidics device (confined channel) (d) log-log plot of in vivo human breast cancer cell (MDA-MB-231) mechanics (elastic modulus, G’, and viscous modulus, G’’) and frequencies (7Hz to 15kHz) in the function of ‘Intravascular’ (n=22), ‘Extravasated’ (n=5), ‘1dpe’ (n=3). (e) log-log plot of in vivo human melanoma (A375SM) mechanics (elastic modulus, G’, and viscous modulus, G’’) and frequencies (7Hz to 15kHz) in the function of ‘Intravascular’ (n=6), ‘Extravasated’ (n=4), ‘1dpe’ (n=3). (f) log-log plot of in vitro human breast cancer cell (MDA-MB-231) mechanics (elastic modulus, G’, and viscous modulus, G’’) and frequencies (7Hz to 15kHz) in the function of ‘In Channel’ (n=38) and ‘Post Channel’ (n=10). Error bars in standard of error. Crossover frequency is assigned for each condition to point out the frequency where G’’ becomes more dominant than G’.

Article Snippet: To establish MDA-MB-231 and A375SM cell lines expressing H2B-mCerulean3, cells were transduced with lentivirus expressing pLentiPGK Hygro DEST H2B-mCerulean3 (Addgene: 90234) diluted 1:50 in growth medium supplemented with 8 μg/ml protamine sulfate and 10 μM HEPES.

Techniques: In Vivo, Injection, Migration, In Vitro

Journal: bioRxiv

Article Title: YAP localization mediates mechanical adaptation of human cancer cells during extravasation in vivo

doi: 10.1101/2023.11.14.567015

Figure Lengend Snippet: (a) Scheme of following up extravasated cancer cell from 3dpf or 0 day-post-extravasation (dpe) to 4dpf/1dpe inside of zebrafish and of brain mechanics where beads were directly injected into brain of Tg(flk:mCherry/MRC1a:EGFP) at 2dpf for measurement at 3dpf/1dpi and 4dpf/2dpi (b) Complex modulus (|G*|) of human breast cancer cells (MDA-MB-231) in the function of ‘Intravascular’ (n=22), ‘Extravasated (0dpe)’ (n=5), and ‘1dpe’ (n=3) and of brain parenchyma at 4dpf (Number of fish = 5, n=154) (c) Complex modulus (|G*|) of human melanoma cells (A375SM) in the function of ‘Intravascular’ (n=6), ‘Extravasated (0dpe)’ (n=4), and ‘1dpe’ (n=3) and of brain parenchyma at 4dpf (Number of fish = 5, n=154) (d) Normalized bar graph of complex modulus for ‘Breast (Intravascular)’, ‘Breast (1dpe)’, ‘Melanoma (Intravascular)’, ‘Melanoma (1dpe)’, and ‘Brain 4dpf’ in respect to ‘Brain 4dpf’ based on 19 different frequencies from 7Hz to 15kHz. **** p<0.0001, paired two-tailed t-tests.

Article Snippet: To establish MDA-MB-231 and A375SM cell lines expressing H2B-mCerulean3, cells were transduced with lentivirus expressing pLentiPGK Hygro DEST H2B-mCerulean3 (Addgene: 90234) diluted 1:50 in growth medium supplemented with 8 μg/ml protamine sulfate and 10 μM HEPES.

Techniques: Injection, Two Tailed Test

Journal: bioRxiv

Article Title: YAP localization mediates mechanical adaptation of human cancer cells during extravasation in vivo

doi: 10.1101/2023.11.14.567015

Figure Lengend Snippet: (a) Scheme of Real-Time Deformability Cytometry (RTDC) measuring physical characteristics of cells under various flow velocities. Examples of cells with various deformation shapes ranging from low to high deformation. (b) Physical characteristics of human breast cancer cell (MDA-MB-231) in terms of deformation and Young’s modulus (kPa) at 0.16 µl/s (N=3; per each replicate more than 1000 cells), 0.24 µl/s (N=3; per each replicate more than 1000 cells), and 0.32 µl/s (N=3; per each replicate more than 1000 cells). **** p<0.0001, paired two-tailed t-tests. (c) Physical characteristics of human melanoma (A375SM) in terms of deformation and Young’s modulus (kPa) at 0.16 µl/s (N=3; per each replicate more than 1000 cells), 0.24 µl/s (N=3; per each replicate more than 1000 cells), and 0.32 µl/s (N=3; per each replicate more than 1000 cells). * p<0.05, ** p<0.01, *** p<0.001, paired two-tailed t-tests. (d) Normalized histogram of human breast cancer cell for Young’s modulus (kPa) at different flow velocities, 0.16 µl/s (n=4906), 0.24 µl/s (n=7398), and 0.32 µl/s (n=9506), with standard deviation (σ, ±68%) (e) Normalized histogram of human melanoma for Young’s modulus (kPa) at different flow velocities, 0.16 µl/s (n= 3954), 0.24 µl/s (n= 6284), and 0.32 µl/s (n= 8442), with standard deviation (σ, ±68%)

Article Snippet: To establish MDA-MB-231 and A375SM cell lines expressing H2B-mCerulean3, cells were transduced with lentivirus expressing pLentiPGK Hygro DEST H2B-mCerulean3 (Addgene: 90234) diluted 1:50 in growth medium supplemented with 8 μg/ml protamine sulfate and 10 μM HEPES.

Techniques: Cytometry, Two Tailed Test, Standard Deviation

Journal: Biomolecules

Article Title: Attachable Hydrogel Containing Indocyanine Green for Selective Photothermal Therapy against Melanoma

doi: 10.3390/biom10081124

Figure Lengend Snippet: In vitro photothermal therapy (PTT) by PAD-ICG hydrogel against melanoma cells. B16 cells were treated with PAD-0.2ICG and irradiated with near-infrared (NIR) laser (1 W/cm 2 for 5 min): ( a ) the average temperature of pure PAD and PAD-0.2ICG after 808 nm laser irradiation at 1 W/cm 2 for 5 min; ( b ) the cell morphology was shown 24 h after the laser irradiation; ( c ) the cell apoptosis and necrosis were analyzed by annexin-V and 4′,6-diamidino-2-phenylindole (DAPI) staining; ( d ) the mean percentage of annexin-V - and DAPI - live cells was shown; ( e ) the intracellular active-caspase 3 levels measured by flow cytometry (left panel) and the mean fluorescence intensity (MFI) of active-caspase 3 are shown; and ( f ) B16, A375P and A375SM were treated with PAD-ICG and irradiated with NIR at 1 W/cm 2 for 5 min. The apoptotic and necrotic cells were analyzed by annexin-V and DAPI staining (left panel). Mean live cells were shown (right panel). Data are representative or show the average of four independent samples (two independent experiments performed with n = 2/group, two-way ANOVA, mean ± SEM), ** p < 0.01.

Article Snippet: The murine melanoma cell line B16 (from American Type Culture Collection, Manassas, VA, USA) and human melanoma cell line A375P and A375SM (both from Korean Cell Line Bank, Seoul, Korea) were cultured in DMEM (Sigma Aldrich) and supplemented with 10% FBS, 2 mM glutamine, 1 M N -(2-hydroxyethyl)piperazine- N ′-(2-ethanesulfonic acid), 100 μg/mL of streptomycin, 100 U/mL of penicillin, and 2 mM 2-mercaptoethanol.

Techniques: In Vitro, Irradiation, Staining, Flow Cytometry, Fluorescence