Fig. 6
p53 plays a critical role in TXN1 mediated effects. A EML cells were transduced with Cas9 control, TXN1 specific CRISPR/Cas9, p53 specific CRISPR/cas9 or both. p53 and P53 downstream signaling pathway were assessed by immunoblot. β-actin was used to assess equal loading. B Cells proliferation rate was assessed using BrdU proliferation assay. C JC-I mitochondrial membrane depolarization was measured D ROS measurement by carboxy-H2DCFDA staining. EML/TXN1KO cells were treated with 250uM NAC and ROS levels were measured. E Cell proliferation. EML/TXN1KO cells were treated with 250uM NAC and cells proliferation rate was assessed using BrdU proliferation assay as described. F In vitro cultured colony forming unit in EML control cells, EML Cas9 control cells, EML TXN1 KO cells and EML TXN1 KO/NAC cells. EML/TXN1KO cells were treated with 250uM NAC, and plated at 2X104/dish in Meth3434 semi-solid medium and incubated for 10-12 days. G JC-1 mitochondrial membrane depolarization was not affected by NAC treatment. EML/TXN1KO cells were treated with 250uM NAC and JC-1 levels were measured. (n = 4 Mean ± SEM or SD)