Fig. 2

Immune response is preserved in xCT^null mice. A Immunization schedule of xCT^wt (WT) and xCT^null (null) BALB/c mice with the empty pVAX1 or the RHuT plasmids. B Percentage of immune cell populations in the blood of WT and null healthy mice. C  Percent of lysis of splenocytes stained with CFSE, pulsed with the target antigen, and injected in mice immunized with pVAX1 or RHuT, as assessed by flow cytometry. D ELISA assay of pre-vax (Pre) and post-vax (Post) sera from vaccinated mice, recognizing Human or Rat ERBB2 extracellular (EC) portion. E Experimental scheme of BM-derived cells culturing and conditioning. F Brightfield microscopy images (10X) of WT or null BM-derived cells cultured for 7 days with CM of 4T1 cells, with or without the addition of 100 µM β-ME. Percentage of G B lymphocytes, H polarized macrophages, I monocytes/mMDSC, J neutrophils/PMN-MDSC and K mature dendritic cells on their respective parent populations indicated in the y-axis legend. L Histogram representation of spleen weight at sacrifice of mice challenged subcutaneously with 10.000 4T1 cells. M Percentage of CD45+ tumor infiltrating cells on total single cells of mice described in panel L. N Growth curves of tumors deriving from 10.000 4T1 cells injected subcutaneously in WT or null mice. O Number of superficial metastases in lungs from mice described in panel N. Number of replicates: Each dot represents a mouse. Statistical analysis: unpaired t test. In panels G-K, t test was performed between xCT^null(null) cells grown in CM without β-ME versus all the other groups. Unpaired t test performed between pre-vax and post-vax values, or between pVAX and RHuT values. * p<0.05; ** p<0.01. Where not indicated, p value is not significant. In some instances, p values are represented in number when not significant. Histograms represent mean values. Error bars (SD) are shown only when n ≥ 5