Figure 6. Nupr1-depletion favours expression of phagocytosis-related genes indispensable for HoCC in PDAC cells.

In the histograms, data are means ± SEM. *p ≤ 0.05, &p ≤ 0.01, £p < 0.005.

1. Heatmap show myeloid- and phagocytosis-related (right) genes expression in control or Nupr1-depleted Panc-1 cells after mock- or TGFβ-treatment for 7 and 24 h.
2. Histograms show the percentage of cannibal mock and TGFβ-treated Panc-1 cells that were previously transfected with the indicated siRNAs, and cultured with or without the CCR1/CCR2 antagonist repertaxin (RTX) or the neutralizing anti-CXCL1 and/or anti-CXCL6 antibodies, as indicated.
3. Panc-1 cells were transfected with a Flag-tagged Nupr1 or an empty vector (EV) control, mock- or TGFβ-treated for 5 h, and chromatin immunoprecipitation (ChIP) was performed using an anti-Flag-tag antibody. The upper panels show occupancy of Nupr1 on CDC42 and CXCL1 promoters, the bottom panels for each promoter show DNA input (10%) before immunoprecipitation of EV and Nupr1-transfected lysates.
4. CDC42 and CXCL1 gene reporter assay. Panc-1 cells were transfected with pGL3-CDC42 or pCXCL1 vectors along with prLucC2 vector and cultured with or without TGFβ. Histogram show normalized luciferase activity indicative of transcription activation.
5. (Left) vinculin (green) and actin (red) immunostaining of Nupr1-depleted BxPC3 cells; (right) histogram shows FACS-based HoCC quantification of BxPC3 Nupr1-depleted and control cells upon TGFβ treatment. Nuclei were counterstained with DAPI.
6. Histogram shows cytofluorometric quantification of HoCC in Nupr1-depleted and control cells that were treated with the indicated combination of TGFβ and the p38 inhibitor SB203580.