Figure 5
Figure 5. Dynamics of Liar expression and localization during Epo-induced erythroid differentiation. (A) Liar mRNA expression during erythroid differentiation. J2E cells were stimulated with Epo and analyzed for Liar mRNA levels by quantitative reverse transcription–polymerase chain reaction (RT-PCR) at 0, 24, and 48 hours after stimulation. (B) Liar and Lyn protein expression during erythroid differentiation. Cells prepared as for panel A were analyzed by Western blot analysis with anti-Liar, anti-Lyn, and anti-Erk2 antibodies. Liar and Lyn levels expressed relative to Erk2. (C) Liar translocates to the nucleus during erythroid differentiation. J2E cells stimulated with Epo were analyzed by confocal microscopy and immunofluorescence staining using anti-Liar and anti-Lyn antibodies. (D) Liar translocates to the nucleus shortly after Epo stimulation. J2E cells stimulated with Epo were analyzed by confocal microscopy and immunofluorescence staining using anti-Liar antibodies. The nucleus was identified by DNA counterstaining with Hoechst 33 258.

Dynamics of Liar expression and localization during Epo-induced erythroid differentiation. (A) Liar mRNA expression during erythroid differentiation. J2E cells were stimulated with Epo and analyzed for Liar mRNA levels by quantitative reverse transcription–polymerase chain reaction (RT-PCR) at 0, 24, and 48 hours after stimulation. (B) Liar and Lyn protein expression during erythroid differentiation. Cells prepared as for panel A were analyzed by Western blot analysis with anti-Liar, anti-Lyn, and anti-Erk2 antibodies. Liar and Lyn levels expressed relative to Erk2. (C) Liar translocates to the nucleus during erythroid differentiation. J2E cells stimulated with Epo were analyzed by confocal microscopy and immunofluorescence staining using anti-Liar and anti-Lyn antibodies. (D) Liar translocates to the nucleus shortly after Epo stimulation. J2E cells stimulated with Epo were analyzed by confocal microscopy and immunofluorescence staining using anti-Liar antibodies. The nucleus was identified by DNA counterstaining with Hoechst 33 258.

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