Fig. 6: DLK1 is a major regulator of ABCB1 and chemoresistance in adrenocortical carcinoma. | Nature Communications

Fig. 6: DLK1 is a major regulator of ABCB1 and chemoresistance in adrenocortical carcinoma.

From: Identification of the Notch ligand DLK1 as an immunotherapeutic target and regulator of tumor cell plasticity and chemoresistance in adrenocortical carcinoma

Fig. 6

A Immunoblot analysis of DLK1 and loading control (α-tubulin) proteins in CU-ACC1 cells with and without DLK1 KO. Four single-cell KO clones are shown. B Immunoblot analysis of NOTCH1 signaling, total NOTCH1 and NOTCH1 intracellular domain (ICD), NE marker synaptophysin (SYP), and loading control (α-tubulin) proteins with and without DLK1 KO in CU-ACC1 cells. Two single-cell KO clones are shown. C Correlation between NOTCH1 and DLK1 expression among TCGA ACC tumors. Pearson correlation coefficients with two-tailed p-values are shown. D DLK1 and NOTCH1 expression in TCGA ACC tumors and normal adrenals. Unpaired t tests were used to calculate two-tailed p-values. Error bar represents mean values ± 95% C.I. E SG3199 cytotoxicity in CU-ACC1 parental and DLK1 KO clones. Cells were treated with SG3199 for 3 days (data representative of n = 4 independent experiments). Error bars represent mean values ± S.E.M. F Flow cytometry histograms assessing ABCB1 in CU-ACC1 cells with and without DLK1 KO. G Immunoblot analysis of DLK1, total NOTCH1 and NOTCH1-ICD, SYP, and α-tubulin proteins in DLK1+ NCI-ACC40, DLK1+ NCI-ACC48 and DLK1- ACC49 PDOs. H Flow cytometry histograms assessing ABCB1 in DLK1 negative NCI-ACC49 PDOs. I Immunoblot analysis of total NOTCH1 (to detect the NOTCH1-ICD plasmid expression), SYP, and α-tubulin proteins in CU-ACC1 cells with and without NOTCH1-ICD overexpression. J Flow cytometry histograms assessing ABCB1 in CU-ACC1 cells with and without N1ICD overexpression. K Correlation between NOTCH1 and ABCB1 expression among TCGA ACC tumors and (L) among GTEx normal adrenal tissues. Pearson correlation coefficients with two-tailed p-values are shown. M Single cell RNA-seq data of ABCB1 expression comparing high NOTCH1 to low NOTCH1 expressing cells from 18 ACC metastatic tumors. Unpaired t tests were used to calculate two-tailed p-values. N Model summarizing the findings of the current study. For immunoblots, experiments were performed two times with similar results. Source data are provided as a Source Data file.

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