JCS -- Jing et al. 120 (7): 1267 Figure IG3

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Fig. 3. Synemin mRNA and protein levels in passage 1 astrocyte cultures from the brain of 2-day-old wild-type, Gfap^-^/^-, Vim^-^/^- and Gfap^-^/^-Vim^-^/^- mice. (A) RT-PCR synemin products from wild type (wt) (lane 2), Gfap^-^/^-Vim^-^/^- (G-/V-) (lane 3), Gfap^-^/^- (G-/V+) (lane 4) and Vim^-^/^- (G+/V-) mice (lane 5). In all genotypes, PCR amplified two bands corresponding to ${alpha}$ - and $beta$ -synemin mRNA. Lane 1 shows molecular mass markers; numbers to the left indicate size in kb. (B) TaqMan real-time PCR amplification curves show no difference in synemin mRNA levels between wild type (full circles) and the different knockout genotypes (triangles). ${Delta}$ Rn, log fluorescence emission intensity of the reporter dye normalized to a passive reference dye. Values are mean ± s.e.m., n=4. Horizontal lines indicate a background fluorescence level. (C) Immunoblots showing synemin, GFAP, vimentin and actin protein levels in passage 1 astrocyte cultures. Synemin was present in astrocytes expressing vimentin, but was undetectable in the absence of vimentin, even when GFAP was present. Actin control blots demonstrate equal protein loading. (D) Immunoblots showing the distribution of synemin, GFAP and vimentin in the detergent-insoluble cytoskeletal fraction (P) and the cytosolic fraction (SN).