Journal: Oncogene
Article Title: Partial truncation of the C-terminal domain of PTCH1 in cancer promotes tumourigenesis by non-canonical activation of a GLI-PI3K loop
doi: 10.1038/s41388-026-03698-9
Figure Lengend Snippet: A Top: EREG , AREG , and EGFR mRNA expression in SCR, C9, and C15 cells by qPCR. Expression levels were normalized to GAPDH mRNA and represented as fold change relative to control SCR cells ( n = 3). Bottom: expression of the same genes in pooled RNA from xenografts derived from SCR and C9 cells. B Representative western blot of EGFR, phospho-Akt, phospho-ERK1/2, total Akt, total ERK1/2, and GADPH in SCR, C9, and C15 cells. C Effect of 10 μM Erlotinib or vehicle control on cell proliferation of SCR, C9 and C15 cells over time ( n = 3). D . Effect of 10 μM Erlotinib or vehicle control (DMSO) on the colony formation ability of SCR, C9 and C15 cells upon culture for 10 days. Left: representative images at the end of the experiment. Right: number of colonies per 1,000 plated cells at day 10 ( n = 3). E Effect of 10 μg/ml Cetuximab or a control IgG on the cell number of SCR, C9 and C15 cells after 72 h treatment ( n = 3). F Effect of 30 nM Trametinib or DMSO vehicle on phospho-ERK1/2 levels in SCR, C9 and C15 cells. G Dose dependent reduction of cell proliferation in SCR, C9, and C15 cells treated with Trametinib for 72 h ( n = 3). All quantitative data represent the mean ± SEM of independent biological repeats (indicated n), performed in technical replicates. ns = not significant; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.
Article Snippet: The following antibodies were used: EGFR (Cell Signaling Technology #2256, WB 1:1,000), AKT (Cell Signaling Technology #4691, WB 1:1,000), phospho- AKT (Ser473) (D9E) XP (Cell Signaling Technology #5012, WB 1:1,000), p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (Cell Signaling Technology #4695, WB 1:1,000), phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (Cell Signaling Technology #4376, WB 1:1,000), Gli1 (Cell Signaling Technology #2643, WB 1:1,000), Phospho-PKA substrate (Cell Signaling Technology #9624, WB 1:1,000), GAPDH-HRP (Proteintech #HRP-60004, WB 1:3,000), Vinculin (Santa Cruz #sc-73614, WB 1:10,000), Poly-ADP ribose Polymerase (Cell Signaling Technology #9542, WB 1:2,000), anti VP16 (Santa Cruz #sc7545, WB 1:1,000),HA-Tag (Invitrogen #26183, WB 1:10,000), goat anti-rabbit IgG HRP (BioRad #172-1019, WB 1:3,000), goat anti-mouse IgG (H L)-HRP conjugate (Bethyl #A90-116P, WB 1:3,000).
Techniques: Expressing, Control, Derivative Assay, Western Blot
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