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. 2014 Apr;1843(4):703–714. doi: 10.1016/j.bbamcr.2014.01.003

Supplementary Fig. 1.

Supplementary Fig. 1

(A) Expression of Gli2 was analyzed in C3H10T1/2, MDA-MB-231 and peripheral blood mononuclear cells (PBMCs) of mouse or human origin using RT-PCR. Expression of SHH, SMO, PTCH1, ULK3 and GLI1 was detected using RT-PCR in PBMCs isolated from three different donors. (B) Mouse Gli2 and human GLI2 proteins were analyzed in C3H10T1/2, PBMCs and MDA-MB-231 cell lysates by immuno-blotting using GN2 antibodies G-20 (sc-20291, Santa Cruz Biotechnology) and AF3635 (R&D Systems). Approximately 60 μg of total protein of whole cell extracts (WCE) or 70 μg of nuclear extract (NE) were loaded per line; Gli2FL — full-length Gli2, * — unspecific bands. (C) C3H10T1/2 cell were treated with Shh or Tgf-βi for 48 h. Expression levels of Gli2 mRNA and protein were analyzed using RT-PCR and WB; *** — p < 0.001; Gli2Rep? — putative repressor form of Gli2. (D) C3H10T1/2 and 3 T3-L1 cells were treated with Shh for 24 h. Proliferation of the cells were tested using ViaLightTM plus kit and normalized with amount of total protein. The data are presented as an average mean of three independent experiments ± S.D., * — p < 0.05.