The E218Q mutant of basigin, but not the E218R mutant, supports lactate transport by MCT1 in Xenopus oocytes

In panel A, oocytes were injected with water (0), or cRNA for MCT1 (1) in the absence or presence (A) of antisense cRNA against basigin and cRNA for rat basigin (B). Western blots are shown for the crude plasma membrane fraction using both MCT1 and basigin antibodies. In panel B rates of L-lactate (30 mM) transport into oocytes measured using BCECF fluorescence are shown as means±SEM of 5–8 separate oocytes. Where indicated, antisense (AS) against basigin as well as the cRNA for WT-, E218Q- or E218R-basigin was co-injected with the MCT1 cRNA.<p><b>Copyright information:</b></p><p>Taken from "The role of charged residues in the transmembrane helices of monocarboxylate transporter 1 and its ancillary protein basigin in determining plasma membrane expression and catalytic activity"</p><p></p><p>Molecular Membrane Biology 2006;23(6):486-498.</p><p>Published online 13 Dec 2006</p><p>PMCID:PMC2409183.</p><p></p

Mutation of Arg or Arg to glutamine or glutamate does not prevent MCT1 from being correctly targeted to the plasma membrane of COS cells

COS cells were co-transfected with MCT1-c-CFP and basigin-c-YFP constructs containing the mutations indicated and live cell imaging performed as described under ‘Methods’. This Figure is reproduced in colour in online.<p><b>Copyright information:</b></p><p>Taken from "The role of charged residues in the transmembrane helices of monocarboxylate transporter 1 and its ancillary protein basigin in determining plasma membrane expression and catalytic activity"</p><p></p><p>Molecular Membrane Biology 2006;23(6):486-498.</p><p>Published online 13 Dec 2006</p><p>PMCID:PMC2409183.</p><p></p

R306K-MCT1 is expressed at the plasma membrane of Xenopus oocytes but is inactive

Details are as given for . Transport measurements are not shown because R306K-MCT1 failed to elicit any lactate transport whether or not WT- or E218R-basigin cRNA was co-injected. This Figure is reproduced in colour in online.<p><b>Copyright information:</b></p><p>Taken from "The role of charged residues in the transmembrane helices of monocarboxylate transporter 1 and its ancillary protein basigin in determining plasma membrane expression and catalytic activity"</p><p></p><p>Molecular Membrane Biology 2006;23(6):486-498.</p><p>Published online 13 Dec 2006</p><p>PMCID:PMC2409183.</p><p></p

R306E-MCT1 and E218R-basigin are not expressed at the plasma membrane of Xenopus oocytes

Oocytes were micro-injected with the cRNA shown and after 72 hours some oocytes were used for immunofluorescence microscopy with the antibody shown (panel A) and others used for membrane preparation followed by SDS-PAGE (20 mg protein) and western blotting with anti-rat MCT1 antibody (panel B). For the western blot, kidney plasma membranes were used as a positive control. The faint band in the water-injected controls represents very slight sample contamination and is only visible because of the over-exposure of the blot to ensure any expressed MCT was detected. Further details are given under ‘Methods’. This Figure is reproduced in colour in online.<p><b>Copyright information:</b></p><p>Taken from "The role of charged residues in the transmembrane helices of monocarboxylate transporter 1 and its ancillary protein basigin in determining plasma membrane expression and catalytic activity"</p><p></p><p>Molecular Membrane Biology 2006;23(6):486-498.</p><p>Published online 13 Dec 2006</p><p>PMCID:PMC2409183.</p><p></p

FRET measurements suggest that mutation of Arg perturbs the interaction of MCT1 with basigin

COS cells were cotransfected with MCT1-c-CFP and basigin-c-YFP constructs containing the mutations indicated and live cell imaging with determination of FRET performed as described under ‘Methods’. Data are presented as means±SEM for the number of observations shown.<p><b>Copyright information:</b></p><p>Taken from "The role of charged residues in the transmembrane helices of monocarboxylate transporter 1 and its ancillary protein basigin in determining plasma membrane expression and catalytic activity"</p><p></p><p>Molecular Membrane Biology 2006;23(6):486-498.</p><p>Published online 13 Dec 2006</p><p>PMCID:PMC2409183.</p><p></p

D302R/R306E-MCT1 is expressed at the plasma membrane of Xenopus oocytes but is inactive

Details are as given for . Transport measurements are not shown because D302R/R306E-MCT1 failed to elicit any lactate transport whether or not WT- or E218R-basigin cRNA was co-injected. This Figure is reproduced in colour in online.<p><b>Copyright information:</b></p><p>Taken from "The role of charged residues in the transmembrane helices of monocarboxylate transporter 1 and its ancillary protein basigin in determining plasma membrane expression and catalytic activity"</p><p></p><p>Molecular Membrane Biology 2006;23(6):486-498.</p><p>Published online 13 Dec 2006</p><p>PMCID:PMC2409183.</p><p></p

MCT2 immunoreactivity in 40 µm sagittal sections through the adult rat cerebellar cortex reveal Purkinje cell immunoreactivity.

<p><b>A:</b> MCT2 immunoreactivity visualised by peroxidase reaction product is deposited in the Purkinje cell dendrites in the molecular layer (ml), Purkinje cell somata in the Purkinje cell layer (pcl), and weakly in the granular layer (gl). <b>B:</b> Anti-MCT2 immunofluorescence staining is prominent in the granular layer (gl). <b>C:</b> Anti-MCT2 immunofluorescence staining outlines Purkinje cell somata in the Purkinje cell layer (pcl) and their proximal dendrites in the molecular layer (ml): the distal dendritic arbour is either weakly stained or unreactive. Scale bar = 50 µm.</p

Echocardiography measurements taken from mice anesthetized with isoflurane.

<p>Data was obtained from M-mode echocardiographic images taken in parasternal short axis mode at the level of the papillary muscles. Data are presented as mean±SEM. There was no statistical significance between the data.</p

I/R injury in isolated hearts with and without the addition of 0.2 µM CsA.

<p>The CsA was added to the buffer 10) Infarct volume quantified from the heart slices stained with TTC. B) Representative heart slices from the normal diet and high-fat diet groups. C) The change in flow rate from pre-ischemia to the end of reperfusion. Data are presented as mean±SEM (n = 5-6 hearts). Data were analyzed using two-way ANOVA with the Bonferroni post-hoc test. *** = P<0.001, ** = P<0.01 vs. normal diet, $$$= P<0.001 and$ = P<0.05 vs. CsA.</p

Characteristics of mice fed normal or high-fat diet.

<p>IPITT = intra-peritoneal insulin tolerance test. Data are presented as mean±SEM. *** = P<0.001 and * = P<0.05 vs. normal diet. Numbers shown in parenthesis indicate number of mice used except for cholesterol, triglycerides and glucose where n refers to number of measurements each containing a pool of 3 samples from 3 separate mice.</p