DOI: 10.5061/dryad.jq2bvq8hd
title: Data from: Tape-assisted fabrication method for constructing PDMS membrane-containing culture devices with cyclic radial stretching stimulation

description: Advanced in vitro culture systems have emerged as alternatives to animal
 testing and traditional cell culture methods in biomedical research.
 Polydimethylsiloxane (PDMS) is frequently used in creating sophisticated
 culture devices due to its elastomeric properties, which allow mechanical
 stretching to simulate physiological movements in cell experiments. We
 introduce a straightforward method that utilizes three types of commercial
 tape—generic, magic, and masking—to fabricate PDMS membranes with
 microscale thicknesses (47.2 µm for generic, 58.1 µm for magic, and 89.37
 µm for masking) in these devices. These membranes are shaped as the bases
 of culture wells and can perform cyclic radial movements controlled via a
 vacuum system. In experiments with A549 cells under three mechanical
 stimulation conditions, we analyzed transcriptional regulators responsive
 to external mechanical stimuli. Results indicated increased nuclear
 YAP/TAZ activity in both confluent and densely packed cells under
 cyclically mechanical strains (Pearson’s coefficient (PC) of 0.59 in
 confluent and 0.24 in dense cells) compared to static (PC=0.47 in
 confluent and 0.13 in dense) and stretched conditions (PC=0.55 in
 confluent and 0.20 in dense). This technique offers laboratories without
 microfabrication capabilities a viable option for exploring cellular
 behavior under dynamic mechanical stimulation using PDMS membrane-equipped
 devices.
description: # Data from: Tape-assisted fabrication method for constructing PDMS
 membrane-containing culture devices with cyclic radial stretching
 stimulation
 [https://doi.org/10.5061/dryad.jq2bvq8hd](https://doi.org/10.5061/dryad.jq2bvq8hd) We have submitted our datasets: measured tape thicknesses (01Commercial tape thickness.xlsx), measured PDMS membrane thicknesses (02PDMS membrane thickness.xlsx), analyzed stains (03Strain.xlsx), analyzed curvature (04Curvature.xlsx), analyzed Pearson's coefficient of different cell densities(05Different cell density_Pearson's coefficient.xlsx), analyzed Pearson's coefficient of confluent cells (06Confluent cells_Pearson's coefficient.xlsx), analyzed Pearson's coefficient of dense cells (07Dense cells_Pearson's coefficient.xlsx), analysis of YAP/TAZ nuclear/cytoplasmic fluorescence intensity (08YAPTAZnuclei&cytoplasm_FluorescenceAnalysis.xlsx) ## Descriptions 01Commercial tape thickness The thicknesses of three different commercial tapes (generic, magic, and masking) were measured under a microscope with image analysis software. Five measured thicknesses were averaged for comparison. 02PDMS membrane thickness PDMS membrane thicknesses made by different tapes were measured under microscope with image analysis software, five samples were measured at three different location (left, right and center) and averaged for comparison. 03Strain Stain values of strained PDMS membranes under four vacuum strengths (-20kPa, -40kPa, -60kPa, and -80kPa) and on different hole diameters of circular frames (250µm, 500µm, 1000µm, and 2000µm) were measured using ImageJ analysis software by calculating the strained line and the unstrained bottom line of the sectional view of the molding PDMS replicas. Five samples of each parameter were averaged for comparison. 04Curvature Curvature values were measured using ImageJ with a plug-in app Kappa by calculating the mean value of all the curvature points. Experimental parameters are four vacuum strengths (-20kPa, -40kPa, -60kPa, and -80kPa) and on different hole diameters of circular frames (250µm, 500µm, 1000µm, and 2000µm). Five samples of each parameter were averaged for comparison. 05Different cell density_Pearson's coefficient Pearson's coefficient of fluorescent images of different cell densities (25000, 50000, 75000,100000,125000,150000 cells per well) were calculated using ImageJ with a plug-in app JACoP BIOP version to quantify the subcellular colocalization of nuclear and YAP/TAZ staining. Three sample images were calculated for each group. 06Confluent cells_Pearson's coefficient Pearson's coefficient of fluorescent images of concluent cell density (25000 cells per well) in three mechanical conditions (static, stretched, and dynamic) were calculated using ImageJ with a plug-in app JACoP BIOP version to quantify the subcellular colocalization of nuclear and YAP/TAZ staining. Seven sample images were calculated for each group. 07Dense cells_Pearson's coefficient Pearson's coefficient of fluorescent images of dense cell density (100000 cells per well) in three mechanical conditions (static, stretched, and dynamic) were calculated using ImageJ with a plug-in app JACoP BIOP version to quantify the subcellular colocalization of nuclear and YAP/TAZ staining. Seven sample images were calculated for each group. 08YAPTAZnuclei&cytoplasm_FluorescenceAnalysis The YAP/TAZ fluorescence intensity of nuclear/cytoplasmic ratio was analyzed using ImageJ. The mean intensity of nuclear YAP/TAZ and non-nuclear YAP/TAZ staining sites were calculated for fluorescence analysis. Three datasets for YAP/TAZ nuclear/cytoplasmic ratio were shown: confluent cells in three mechanical conditions, dense cells in three mechanical conditions, and different cell densities.