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. 2011 Jan 7;6(1):e14500. doi: 10.1371/journal.pone.0014500

Figure 3. Gadd45a localization in nuclear speckles is RNase sensitive.

Figure 3

A, Scheme of detergent extraction. B, RKO cells expressing EGFP-Gadd45a were subjected to detergent extraction with or without RNaseA treatment followed be Western blot analysis of the indicated proteins. A representative experiment out of three performed is shown. C, D Immunofluorescence confocal microscopy of detergent-extracted RKO cells. Cells were transfected with N-EGFP-Gadd45a and stained with antibodies against SC35 (C) and p68 (D). E, cells were treated as in B, but subjected to RNase treatment after extraction and before fixation. F, Statistical analysis for localization of EGFP-Gadd45a and SC-35 in nuclear speckles in cells with and without RNaseA treatment (n = 35 cells; n = 3 experiments; a representative experiment is shown). G, H Immunofluorescence confocal microscopy of endogenous Gadd45a in UV irradiated detergent-extracted HEK293T cells. Cells were stained with antibodies against Gadd45a and SC35. In (H), cells were subjected to RNase treatment after extraction and before fixation. I, Statistical analysis for localization of UV inducible Gadd45a in nuclear speckles in HEK293T cells with and without RNaseA treatment (n = 50 cells).