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. 2020 May 7;12(5):1175. doi: 10.3390/cancers12051175

Figure 5.

Figure 5

Sens and CPR TYK-nu showed different behaviors under shaking conditions. (A,B) Sens TYK-nu and CPR TYK-nu were seeded onto HA and FN matrices and allowed to grow into confluent tight monolayers. After 6 h on an orbital shaker (130 RPM), CPR TYK-nu underwent the formation of spheroids (B), whereas Sens TYK-nu maintained their original culture features. (A) Cells were evaluated under a Leica DMIL inverted microscope (Leica Microsystem, Milan, Italy), and images were collected using a Canon Powershot A640 digital camera (Canon, Tokyo, Japan). Original magnification: 200×. (C–E) A cytofluorimetric characterization of Sens (dashed line) and CPR (black line) TYK-nu was performed in terms of β1-integrin (C), HA binding protein (D)—also known as gC1qR/p33—and CD44 (E) expression. The fluorescence intensity of the cells incubated with primary antibodies was compared with unrelated straining (gray line). The expression of CD44 by CPR TYK-nu was completely absent. These data were also confirmed by evaluating CD44 mRNA expression level through real-time qPCR experiments (F), which highlighted an increased level of CD44 when the cells were seeded onto FN as compared to on HA. ** p < 0.01.