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Figure 2. FGFR1 overexpression sustains Akt and Erk pathway activation in alpelisib-treated MCF-7/FGFR1 cells. (A) MCF7/C and MCF7/FGFR1 cells were treated with alpelisib (Alp) at the indicated concentrations for 24 h. Protein levels of p-Akt, Akt, p-Erk1/2, Erk1/2, p-S6K, S6K, p-Rb, Rb, and Cyclin D1 in each sample were detected by Western blotting, with GAPDH used as an internal control; (B) Densitometry analysis of phosphorylated/active signaling markers (p-Akt, p-Erk, p-S6K, p-Rb) and Cyclin D1 in MCF-7/C (M) and MCF-7/FGFR1 (MF) cells treated with alpelisib. Band intensities in (A) were quantified (ImageJ), normalized to loading controls (GAPDH) and total protein levels based on three repeats. ^*P < 0.05; ^**P < 0.01, based on comparisons of the marker between MCF-7/FGFR1 and MCF-7/C cells under the same treatment conditions.