Fig. 2: The androgen promotes the MICA/B shedding in melanoma cells, leading to the reduction of the NKG2D expression in NK cells.

A, B Frequency of MICA/B in untreated (white bars) or R1881-treated (black bars) AMM16 cells. B Data from three independent experiments are represented as mean ± SD. n = 3; **p-value < 0.01. C AMM16 were untreated or treated for 6 h with R1881, in absence or presence of bicalutamide (Bic). When indicated, cells were pretreated with MG132. WB using the indicated antibodies was done. D Quiescent AMM16 cells on coverslips were left untreated or treated for 6 h with R1881. They were stained for MICA/B. Images captured by confocal microscope show the staining of MICA/B (green) and nuclei (blu). Merged images are presented in the right panels. They are representative of three independent experiments. Bar, 10 μm. E Cells were unstimulated or stimulated with R1881, in the absence or presence of Bic for 6 h. sMICA was assayed and quantified by ELISA. Graph is representative of three independent experiments; *p-value < 0.05. F, G NK cells were untreated or treated with AMM16 cells derived CM, as indicated. F WB with the antibodies against the indicated proteins was done. G NK cells were stained as reported in Methods section. Images captured by confocal microscopy show the staining of NKG2D (red) and nuclei (blu). Merged images are presented in the right panels. They are representative of three independent experiments. Bar, 2.5 μm.