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. 2018 Sep 11;10(7):1003–1017. doi: 10.1080/19420862.2018.1503904

Figure 2.

Figure 2.

HER2-dependent T-DM1 internalization is blocked in T-DM1R cells. (A) The levels of endogenous HER2 in the WCL of JIMT1 parental and T-DM1R cells were analyzed by Western blot. (B) Fluorescent immunostaining images showing HER2 in JIMT1 parental and T-DM1R cells. Nucleus was stained with DAPI. Scale bar, 20 µm. (C and D) JIMT1 parental cells were incubated with 100 µg/ml T-DM1 at 37°C for 1 hr and 6 hrs, and subsequently fixed for fluorescent immunostaining of T-DM1 using DyLight 488-conjugated anti-human IgG antibody (green in C and D), HER2 (red in C) and LAMP-1 (red in D). Nucleus was stained with DAPI (blue in C and D). Scale bar, 20 µm. (E and F) JIMT1 T-DM1R cells were incubated with 100 µg/ml T-DM1 at 37°C for 1 hr and 6 hrs, and subsequently fixed for fluorescent immunostaining of T-DM1 using DyLight 488-conjugated anti-human IgG antibody (green in E and F), HER2 (red in E) and LAMP-1 (red in F). Nucleus was stained with DAPI (blue). T-DM1 were not co-localized with either HER2 or LAMP-1. Scale bar, 20 µm.