Fig. 1. Arhgef4 knockout by gene disruption. (A) Chromosomal region of Arhgef4 gene and diagram of vector for knockout (modified information provided by KOMP). FRT: flippase recognition target, lacZ: β-galactosidase, neo: neomycin, loxP: locus of X-over P1. (B) RT-PCR analysis. Five micrograms of total RNA was transferred to cDNAs of Arhgef4 and β-actin by reverse transcription. The 3’untranslated regions of their cDNAs near the poly (A) tail or coding regions were amplified by PCR. (C) Forty micrograms of brain lysates from WT mice (Arhgef4+/+), Homo mice (Arhgef4-/-), or rats, and 10 μg of human embryonic kidney (HEK) cells were separated by 8% acrylamide gel. The western blotting assay was performed using rabbit polyclonal anti-Arhgef4 antibodies (specific for N-terminal regions of mouse Arhgef4; see Materials and Methods). After the detection of Arhgef4, the blots were stripped and subsequently reprobed with anti-α-tubulin antibody.
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