Fig. 1. CALCB is an EWSR1-FLI1 target gene specifically expressed in Ewing sarcoma (EwS).

a Analysis of CALCB expression in EwS (n = 50) and normal tissues (71 tissue types, n = 929 samples). Data are represented as box plots in log₂ scale. Horizontal bars indicate median expression levels, boxes the interquartile range. Whiskers indicate the 2.5th and 97.5th percentiles, respectively. Unpaired two-tailed Student’s t test. b Analysis of CALCB expression in EwS (n = 50) and different (pediatric) tumors (49 types, n = 1 699 samples). Publicly available microarray data are represented as dot plots in log₂ scale with mean and SEM. Each dot represents one sample. The number of samples is given in parentheses. EwS highlighted in blue color. c Time-course analysis of CALCB and EWSR1-FLI1 expression in A673/TR/shEF1 EwS cell harboring a dox-inducible shRNA against EWSR1-FLI1 by qRT-PCR in vitro after dox application. Given are mean expression levels and SEM (n = 7). d Analysis of CALCB expression in xenografts derived from A673/TR/shEF1 cells with/without dox treatment for 96 h in vivo. Gene expression levels were determined by Affymetrix Clariom D microarrays as previously described²⁹. Expression levels are shown in log₂ scale; horizontal bars indicate mean expression level (n = 3); unpaired two-tailed Student’s t test. e Analysis of CALCB expression in a published dataset (GSE64686)⁴⁰ with ectopic dox-inducible EWSR1-FLI1 expression in human embryoid bodies. Data were generated on Affymetrix HG-U133Plus2.0 microarrays and normalized simultaneously by RMA and brainarray CDF (ENTREZg; v21). Horizontal bars indicate mean expression levels; unpaired two-tailed Student’s t test. **P ≤ 0.01; ***P ≤ 0.001