Journal: Biomaterials
Article Title: Integrin α 6 and EGFR Signaling Converge at Mechanosensitive Calpain 2
doi: 10.1016/j.biomaterials.2018.05.056
Figure Lengend Snippet: (a) Clustering of PCR data on genes expressed after 24 hours of culture on gels functionalized with 10μg/cm2 collagen 1 (data in Fig. 1a). (b) Calpain 2 activity during adhesion was measured by evaluating the ratio of autolyzed/full length calpain 2 via western blot in both the adhered and suspended cell fractions during the first 60 minutes, or just the adhered fraction after 24 hours. N=2. (c-d) Cell area was quantified during adhesion in the presence of an ERK1/2 inhibitor or calpain 2 inhibitor on 1 kPa (c) and 41 kPa (d) gels. N=2, n=50. (e) Cell migration speeds quantified the presence of an ERK1/2 or calpain 2 inhibitor. N=2, n=48. (f) Representative western blot for full length and autolyzed calpain 2, with loading control GAPDH. In order of lanes from left to right: 1 kPa with collagen 1, 41 kPa with collagen 1, TCPS, 1 kPa with collagen 1 + 100ng/mL soluble EGF for 10 minutes, 41 kPa with collagen 1 + 100ng/mL soluble EGF for 10 minutes, TCPS + 100ng/mL soluble EGF for 10 minutes, 1 kPa with collagen and laminin, 41 kPa with collagen and laminin.
Article Snippet: For inhibitor conditions, cells were treated 2 hours prior to the start of imaging with 10 μg/mL calpain inhibitor IV, 20 μM ERK inhibitor {"type":"entrez-nucleotide","attrs":{"text":"FR180204","term_id":"258307209"}} FR180204 , 20 μM lapatinib, or 10 ng/mL EGF (R&D Systems).
Techniques: Activity Assay, Western Blot, Migration