Fig. 1

GLI1 expression and primary cilia in RMS and EWS cell lines. a Expression of GLI1 protein by Western blot in RMS and EWS cell lines. Size markers in kilodaltons (kD) are included. GLI1-amplified RMS-13 rhabdomyosarcoma cells or GLI1- amplified Rh30 cells were used as positive controls. The GAPDH control was included to show comparability of protein loading among lanes. The blots have been cropped and full-length blots are presented in Supplementary Figure 1A. b EWS cell lines that were established following recurrence (CHLA258 and TC71) had significantly higher GLI1 expression compared with cell lines established at the time of diagnosis (CHLA9, CHLA10, and TC32) by qRT PCR. c Immunofluorescence microscopy for primary cilia in RMS and EWS cell lines. Cell line names are indicated above the images. Green = pericentrin or pericentrin2 (centriole component), red = acetylated alpha tubulin (primary cilia component), and blue = DAPI nuclear staining. Primary cilia, indicated by arrows, were seen in RD, Rh30, Rh41, CHLA9, CHLA10, CHLA258, and TC32 cells. NIH3T3 cells were used as a positive control. White dotted lines indicate cell borders. Scale bars are shown. Microsoft Exel and Adobe Photoshop were used to prepare Figure 1