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. 2022 May 26;17(5):e0268300. doi: 10.1371/journal.pone.0268300

Fig 2. Matrix stiffness modulates chemotherapeutic resistance to gemcitabine but this is not attributed to classical EMT.

Fig 2

(A) MCF-7 cells were cultured on 500 Pa and 4 kPa hydrogels for 24 hrs prior to the addition of chemotherapeutic agents as indicated; 5-fluroruracil (5-FU), hydroxyurea (HU), and gemcitabine. After 72 hrs cell viability was measured by alamarBlue assay. The means and SEMs are plotted for >3 independent repeats. Significance was calculated using a Student’s paired t-test. (B) E-cadherin and β-catenin visualised by immunofluorescence in MCF-7 cells cultured on 500 Pa and 4 kPa stiffness hydrogels for 24 hrs. (C) Western blot for E-cadherin and a GAPDH control of whole cell extracts from MCF-7 cells cultured on 500 Pa and 4 kPa stiffness hydrogels for 72 hrs. (D) Immunofluorescent visualisation of YAP in MCF-7 cells cultured on 500 Pa and 4 kPa stiffness hydrogels for 24 hrs. (E) Nuclear YAP intensity (integrated density) measured in approximately 100 nuclei for each condition for each of 3 independent repeats. The median and individual cells from pooled data are plotted. Significance was calculated using a Mann-Whitney U-test.