ABCA12 Antibody NB100-93466 from Novus Biologicals, a Bio-Techne Brand

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Science advances
December 01 2023

Fig 1: Lipidomic analyses reveal that ABCA12 reduces intracellular ceramide abundance to maintain cancer stemness.(A) Lipid species significantly down-regulated in SOX9high cells (fold change > 2) in C3/TAg;Sox9-GFP tumor (n = 5). (B) Level of ceramide lipids in Sox9neg, Sox9low, and Sox9high C3/TAg tumor cells. (C) Lipid pathway enrichment analyses of the lipids identified in (A). AGE, advanced glycation end products; RAGE, receptor for AGE. (D) Flow cytometry measuring the abundance of ceramide in the SOX9neg, Sox9low, and Sox9high C3/TAg tumor cells (n = 7). (E) Abundance of ceramide in the SOX9neg, Sox9low, and Sox9high PyMT tumor cells (n = 5). (F) Abundance of ceramide species in sgNT and sgAbca12 C3/TAg organoids (n = 3). (G) Ceramide levels in C3/TAg organoids treated with dimethyl sulfoxide (DMSO) or D-NMAPPD for 5 days as measured by flow cytometry (n = 5). (H) Effect of C6-ceramide or D-NMPAAD on C3/TAg organoid formation (n = 6). (I) Ceramide levels in PyMT organoids treated with DMSO or D-NMAPPD (10 μM) for 5 days (n = 5). (J) Effect of C6-ceramide or D-NMPAAD on PyMT organoid formation (n = 3). (K and L) Organoid-forming efficiency of C3/TAg [(K), n = 5] or PyMT [(L), n = 3] tumor cells pretreated with D-NMAPPD (20 μM) for 3 days. (M) Ceramide levels in C3/TAg organoids treated with DMSO or Fumonisin B1 (FB1; 10 μM) for 5 days (n = 4). (N and O) Effect of FB1 on organoid formation by SOX9neg/low C3/TAg [(N), n = 2] or SOX9neg/low PyMT [(O), n = 3] tumor cells. All data are represented as means ± SEM. P values were determined by one-way ANOVA [(D) and (E)], or with Tukey’s test [(H), (J), (N), and (O)], or paired two-tailed t test [(G), (I), and (K) to (M)]. ***P < 0.001, **P < 0.01, and *P < 0.05.
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Science advances
December 01 2023

Fig 2: ABCA12 lipid transporter is up-regulated in SOX9high cells and required for CSC activity.(A) Volcano plot of RNA-seq comparing SOX9low and SOX9high cells from C3/TAg;Sox9-GFP tumors (n = 4). (B) Sox9 and Abca12 mRNA levels in pLVX-ctrl and pLVX-SOX9 transduced C3/TAg organoids (n = 4). (C) UMAP showing SOX9 and ABCA12 levels in tumor cells in patients with breast cancer (BRCA) (GSE176078). (D) ABCA12 expression in PDX tumors treated with vehicle or cisplatin as in Fig. 4T (H39: n = 8 each group; H17: vehicle, n = 8; cisplatin, n = 9). (E) Organoid-forming efficiency of sgNT and sgAbca12 C3/TAg;Sox9-GFP tumor cells (n = 4). (F) Tumor-initiating cell frequency of sgNT and sgAbca12 C3/TAg;Sox9-GFP organoid cells, as determined by ELDA (n = 2). (G) Growth of sgNT and sgAbca12 C3/TA;Sox9-GFP tumor (sgAbca12/cisplatin, n = 3; other groups, n = 4). (H and I) Sox9-GFP MFI (H) and percentage of Sox9-GFPhigh cells (I) in sgNT and sgAbca12 C3/TAg;Sox9-GFP tumors treated in (G). (J) SOX9 Western blot of the indicated cells. (K) Organoid-forming efficiency of control (sgNT) or Abca12-overexpressing (CRISPRa-Abca12) C3/TAg;Sox9-GFP tumor cells. (L) Effect of cisplatin on organoid formation in control (sgNT) or Abca12-overexpressing (CRISPRa-Abca12 #b) C3/TAg;Sox9-GFP tumor cells. IC50, median inhibitory concentration. (M) Organoid-forming efficiency of the indicated C3/TAg;Sox9-GFP tumor cells. (N) Organoid-forming efficiency of sgNT or sgABCA12-transduced human cell lines. (O) Doxorubicin dose response of sgNT and sgABCA12-transduced breast cancer cells in organoid culture. All data are represented as means ± SEM. P values were determined by unpaired two-tailed t test [(B), (D), (E), and (N)], one-way ANOVA with Dunnett’s (K) or Tukey’s test [(H) to (I) and (M)], Chi-square test in ELDA (F), ordinary two-way ANOVA [(L) and (O)], or two-way RM ANOVA (G). ****P < 0.0001, ***P < 0.001, **P < 0.01, and *P < 0.05.