Fig. 1: The amount of FGFR2 in the lipid rafts is increased after its ligand-dependent activation. | Cell Death & Disease

Fig. 1: The amount of FGFR2 in the lipid rafts is increased after its ligand-dependent activation.

From: Role of lipid rafts in the FGFR2c-mediated oncogenic signaling by involvement of TRPA1 channel in pancreatic ductal adenocarcinoma cells

Fig. 1: The amount of FGFR2 in the lipid rafts is increased after its ligand-dependent activation.The alternative text for this image may have been generated using AI.

PANC-1 cells transiently transfected with pCI-neo FGFR2c expression plasmid (PANC-1 FGFR2c) were left untreated or stimulated with FGF2 for 1 h 4 °C, to trigger receptor activation, impairing its internalization. A Western blot analysis, performed using, shows that the 120 KDa band corresponding to the molecular weight of FGFR2 is more evident in cells transfected with the FGFR2c variant. The equal loading was assessed using anti-beta actin (ACTB) antibody. For densitometric analysis, the values from three different experiments (n = 3) were normalized, expressed as mean values ± SD and reported in a graph as fold increase with respect to the control value. Student t test was performed, and significance levels have been defined as p < 0.05: **p < 0.01. B Quantitative immunofluorescence analysis was performed using anti-FGFR2 bek H80 antibody, which recognizes the extracellular portion of the receptor, and CTxB-FITC, which binds the monosialic ganglioside type 1 (GM1). Nuclei were stained with DAPI. Colocalization analysis was performed by scanning cells with an ApoTome System (Zeiss) and analyzing the images from two independent experiments (n = 2) with the Axiovision software (Zeiss). The treatment with FGF2 increases the colocalization of FGFR2c with GM1 and changes the plasma membrane distribution of the receptor from continuous to dotted. The colocalization was expressed as mean percentage ± SD. Student t test was performed, and significance levels have been defined as p < 0.05: ***p < 0.001. Bars: 10 μm C PANC-1 FGFR2c cells were lysed to obtain TX100 soluble and insoluble fractions as reported in Materials and Methods. The efficiency of the fraction extraction was assessed by checking the distribution of the lipid raft marker flotillin. Western blot analysis shows a significant increase of FGFR2c amount in the insoluble fraction after FGF2 stimulation. Densitometric analysis was performed as reported: *p < 0.05.

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