Extended Data Figure 8: Investigations into why GLI1 levels are elevated in FRII cells.
From: The sonic hedgehog factor GLI1 imparts drug resistance through inducible glucuronidation
a, Analysis of expression of a subset of hedgehog signalling pathway proteins. Western blots are probed as indicated and are representative of three independent experiments. Tubulin and Hsp90 provided loading controls. Patched 1 (PTCH1) was the most significant change. PTCH1 is 210 kDa, with an often observed degradation product at 170 kDa. IHH, Indian hedgehog; SHH, sonic hedgehog, Smo, smoothened. b, GLI1 DNA methylation. CpG methylation was interrogated on bisulphite-converted DNA from GLI1 promoter region and first exon. The amplicon covered 29 CpGs, 25 of them located within a CpG island. DNA CpG methylation is shown as per cent methylation for FaDu (top) and FRII (bottom) cells. There was no difference observed between the cell lines. c, Antibody controls. Analysis of GLI1 and pan-UGT1A antibodies as a function of RNAi-mediated knockdown of these proteins as indicated. Note that UGT1A family members have approximately the same molecular weight. Results are representative of at least three independent experiments.
