Figure 4: The ALKBH4-Actomyosin interaction is mediated by actin.
From: ALKBH4-dependent demethylation of actin regulates actomyosin dynamics
(a) Seventy-two hours after induction with 0.5 μM 4-OHT Control Alkbh4+/+Cre (Alkbh4+/+) and Alkbh4L/LCre (Alkbh4Δ/Δ) MEF cells were lysed and subjected to NM II immunoprecipitation. Immunoprecipitates were analysed by immunoblotting with the indicated antibodies. (b) Forty-eight hours after transfection with the indicated siRNAs, 293T cells were lysed and subjected to NM II immunoprecipitation and subsequently immunoblotted with the indicated antibodies. (c) Forty-eight hours after transfection with the indicated siRNAs and DNA constructs, 293T cells were lysed and subjected to Flag immunoprecipitation. Immunoprecipitates were analysed by immunoblotting with the indicated antibodies. (d) Forty-eight hours after transfection with the indicated DNA constructs, 293T cells were treated with 5 μM Blebbistatin for 5 h, lysed and subjected to HA immunoprecipitation. Immunoprecipitates were analysed by immunoblotting with the indicated antibodies. (e) Forty-eight hours after transfection with the indicated DNA constructs, 293T cells were treated with 1 μg ml−1 cytochalasin D (CCD) for 1.5 h, lysed and subjected to Flag immunoprecipitation. Immunoprecipitates were analysed by immunoblotting with the indicated antibodies. Full-length blots in all panels are presented in Supplementary Fig. S7.
