Fig. 1: Effects of AGX51 on ID proteins.

a Western blot for ID1 on whole-cell lysates from 4T1 cells treated with 0–80 µM AGX51 for 24 h. b Western blot for ID1, ID3, and ID4 on whole-cell lysates from 4T1 cells treated with 40 µM AGX51 for 0–72 h. Quantification of western blot bands is shown in Supplementary Fig. 1a. c Western blot for ID1 on whole-cell lysates from the PDX cell line IBT treated with 40 µM AGX51 for 0–24 h. d Western blot for ID1 and ID3 on whole-cell lysates from 4T1 cells treated with 40 µM AGX51 for 24 h, after which compound was washed off and cells were collected for 0–48 h post compound removal. e qRT-PCR analysis for Id1 in 4T1 cells treated with 40 µM AGX51 for 2, 4, or 48 h. Mean fold difference with error bars showing standard error of the mean (SEM) of technical triplicates. The experiment was performed at least three times. f Electrophoretic mobility shift assay (EMSA) on whole-cell lysates from 4T1 cells treated with 40 µM AGX51 for 24 h. Arrow indicates binding to DNA. g EMSA on whole-cell lysates from 4T1 cells treated with 40 µM AGX51 for 1 h, with corresponding western blot for Id1 shown to the right of the EMSA blot. Arrow indicates binding to DNA. Tubulin/actin are used as protein loading controls. See also Supplementary Fig. 1.