Fig 1: PDGFR-ß interacts with FAK and TLN1 in ECs. a Strategy for identifying PDGFR-ß binding proteins using LC-MS/MS analysis. Bone morrow endothelial cells (BMECs) treated with PDGF-BB (300 ng·mL-1) for 24 h were subjected to immunoprecipitation (IP) with an antibody against PDGFR-ß followed by LC-MS/MS analysis. b The PDGFR-ß binding proteins FAK and TLN1 were identified with LC-MS/MS analysis. c PDGFR-ß was immunoprecipitated from BMECs with an anti-PDGFR-ß antibody. The presence of FAK, TLN1, and PDGFR-ß in these immunoprecipitates was evaluated with immunoblotting. Western blot analysis of FAK, TLN1, and VEGF in the BMECs treated with different PDGF-BB doses (0-300 ng·mL-1) (d) or for different times at 0 h–36 h (e). Western blot analysis of FAK, TLN1, and VEGF in the BMECs treated with FAK inhibitors (f) or TLN1 siRNA (g)
Fig 2: Mechanical cues promoted PKCa recruitment by SDC4. (A) Immunofluorescence of endogenous PKCa and SDC4 in TNBC cells treated with the hypotonic buffer for 300 s. Cells were transfected with on-targeting siRNA or talin 1 siRNA. PI3K inhibitor (LY-294002, 30 µM) or Cytochalasin D (2 µM) was pre-incubated 30 min before the experiment. (B) and (C) Membrane translocation of PKCa in hypo-osmotic-treated TNBC cells, tested by western blot. ATP1A1 as membrane protein loading control, ß-actin as cytoplasm protein loading control. Quantification of PKCa/ATP1A1 was showed (mean ± SD, n = 3. One-way analysis of variance was used for single-factor sample comparisons. *P ATP1A1 as membrane protein loading control, ß-actin as cytoplasm protein loading control. (D) MDA-MB-231 cells expressing srGAP2-M-cpstFRET probes in stiff-directed matrix. CFP/FERT of the forty parts represented srGAP2 tension along with the moving direction.
Supplier Page from Abcam for Anti-Talin 1 antibody [97H6]