ZFIN Image: Iwase et al., 2007, Fig. S6

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Figure Caption

Fig. S6 Comparable Expression of RNAi-Resistant SMCX in Rat Cerebellar Granule Neurons
(A) Expression of RNAi-resistant SMCX in 293T cells. Equal amount of pcDNA3.0 vector or expression plasmid carrying RNAi-resistant WT, H514A, or F642L SMCX was transfected into 293T cells, and resulting cell lysates were analyzed by Western blotting using anti-HA monoclonal antibody. β-actin was used as a loading control. All constructs gave comparable expression level.
(B) In parallel to the rescue experiment in Figure 7E, the transfected neurons were subjected to immunocytochemistry with a rabbit anti-GFP polyclonal antibody and mouse anti-HA monoclonal antibody. GFP and HA-tagged SMCX were visualized using Alexa 488 anti-rabbit IgG (a, d, g, and j) and Alexa 594 anti-mouse IgG (c, f, I, and j), respectively. DAPI was used for all counterstaining of nuclei (b, e, h, and k). Three rescue constructs, which carry HA-tagged WT, H514A, or F642L SMCX, exhibit comparable expression level in the nuclei of GFP-positive granule neurons (f, i, and l).

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Reprinted from Cell, 128(6), Iwase, S., Lan, F., Bayliss, P., de la Torre-Ubieta, L., Huarte, M., Qi, H.H., Whetstine, J.R., Bonni, A., Roberts, T.M., and Shi, Y., The X-linked mental retardation gene SMCX/JARID1C defines a family of histone H3 lysine 4 demethylases, 1077-1088, Copyright (2007) with permission from Elsevier. Full text @ Cell