Fig. 5: c-MYC is upregulated by KTN1 knockdown and directly binds to the promoter region to transactivate CCDC40. | Cell Death & Disease

Fig. 5: c-MYC is upregulated by KTN1 knockdown and directly binds to the promoter region to transactivate CCDC40.

From: Kinectin1 depletion promotes EGFR degradation via the ubiquitin-proteosome system in cutaneous squamous cell carcinoma

Fig. 5

A Schematic diagram for the positions of two predicted promoters in the CCDC40 gene. B, C A431 cells were transfected with siKTN1. Total cell lysates were subjected to ChIP-qPCR assay and immunoblotting. TSS: transcription start site. qPCR results represent the means ± SD. Representative of results from three biological replicates. D Luciferase reporter assay of the predicted promoter #2. Data represent the means ± SD. Representative of results from three biological replicates. E, F A431 cells were transfected with siKTN1 or co-transfected with siKTN1 and sic-Myc. Total cell lysates were subjected to ChIP-qPCR assay and immunoblotting. qPCR results represent the means ± SD. Representative of results from three biological replicates. G Schematic diagram for the predicted interaction site (red underlined bases) of c-Myc (c-Myc binding motif) in CCDC 40 promoter #2. H Electrophoretic mobility shift assay (EMSA) to detect the direct interaction between the c-Myc binding motif and purified c-MYC protein. Representative of results from three biological replicates. I Nuclear lysates from A431 and HSC-5 cells were subjected to EMSA to detect the direct interaction between the c-Myc binding motif and nuclear protein. Anti-c-Myc antibody was added for supershift of the binding complex. Representative of results from three biological replicates.

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