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. 2011 Aug 16;30(20):4211–4222. doi: 10.1038/emboj.2011.303

Figure 4.

Figure 4

Proline 33 of ADAR2 is required for the Pin1 effect on ADAR2 nuclear localization. (A) Normal localization of ADAR2 and Pin1. Immunofluorescence of HeLa cells cotransfected with GluR2 B13 minigene, FLAG–ADAR2 and HA–Pin1 stained with (i) DAPI, (ii) anti-HA–Pin1 (green), (iii) anti-FLAG–ADAR2 (red), (iv) Merge of DAP1 and FLAG and (v) merge of all three images. (B) Nuclear localization of ADAR2 depends on Proline 33. Immunofluorescence of HeLa cells cotransfected with GluR2 B13 minigene, FLAG–ADAR2P33A and HA–Pin1 stained with (i) DAPI, (ii) anti-HA–Pin1 (green), (iii) anti-FLAG–ADAR2P33A (red), (iv) Merge of DAP1 and FLAG and (v) merge of all three images. All photographs were taken at the same exposure. Scale bar, 10 μm. (C) Nucleo-cytoplasmic fractionation of wild-type and ADARP27A and ADAR2P33A mutants. Immunoblot analysis with anti-FLAG antibody of nuclear and cytoplasmic fractions of HeLa cells transfected with FLAG–ADAR2 (lanes 1 and 2), ADARP27A (lanes 3 and 4), ADAR2P33A (lanes 5 and 6). (Lower panels) Immunoblot of fractionated HeLa cells with tubulin as a cytoplasmic marker and HP1α as a nuclear marker.