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. 2021 Apr 22;13(9):2022. doi: 10.3390/cancers13092022

Figure 3.

Figure 3

Downregulation of EGFR in response to DLL4 binding in H1975 and LC31 parental cells and modulation of Sp1 and p53 transcription factors. (A) Dose-dependent decrease of EGFR levels in response to the DLL4 stimulation for 72 h of H1975 parental cells, and the concomitant activation of Notch1 signalling cascade and levels of p53 and Sp1. (B) Direct or indirect interaction between Sp1 and p53 in whole cell lysates from H1975 and H1975-TS assessed by immunoprecipitation with Sp1 or p53 antibodies. (C) Levels of EGFR, Notch1, RBPSUH, Sp1, p53, and EMT markers in parental LC31 and LC31 tumor spheres (TS). (D) Dose-dependent decrease of EGFR levels in response to DLL4 stimulation for 72 h of parental LC31 cells, concomitant activation of Notch1 signalling cascade and levels of p53 and Sp1. (E) H1975 and LC31 parental cells were transfected with siRNA scrambled (CTR) Sp1- and p53-targeted siRNAs, either alone or in combination for 72 h, and levels of EGFR and Notch1 were determined by western blot analysis. At least three independent experiments were performed. GAPDH was used to ensure equal loading.