Exploratory experiment designed to discover new patterns targeting TFEB, Thbs1 in primary cardiac fibroblasts from R26-LSL-TFEB+/+; Acta2-cre and R26-LSL-TFEB+/+ mice. Primary outcome: protein expression levels and cell migration capacity
Cardiac fibroblasts were extracted from adult R26-LSL-TFEB+/+; Acta2-cre and control R26-LSL-TFEB+/+ mice to examine the effects of TFEB overexpression on protein expression and cellular behavior. Western blot analysis was performed to detect protein levels of integrin, CD47, CD36, Thbs1, p-paxillin, vinculin, P-FAK, and α-SMA. Cell migration capacity was assessed using wound healing and Transwell migration assays. The results showed elevated Thbs1 protein levels in TFEB-overexpressing cardiac fibroblasts, along with modulation of focal adhesion complex proteins including reduced activation of p-paxillin, vinculin, and P-FAK. These changes were associated with altered cellular adhesion dynamics and migration properties of cardiac fibroblasts.
Cardiac fibroblast isolation, Western blot analysis for multiple proteins, wound healing assay, Transwell migration assay
TFEB overexpression would modulate focal adhesion proteins and reduce fibroblast migration
Significant changes in protein expression and migration capacity between TFEB overexpressing and control cells
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