Supplementary Figure 3: Membrane surface trafficking of Magneto2.0 is impaired in barrel cortical cells in vivo. | Nature Neuroscience

Supplementary Figure 3: Membrane surface trafficking of Magneto2.0 is impaired in barrel cortical cells in vivo.

From: Revaluation of magnetic properties of Magneto

Supplementary Figure 3

(a) Schematic drawing outlines the design of biochemistry analysis of Magneto2.0 and TRPV4 (both of which are FLAG tagged) expressed in the intact mouse barrel cortex with lentivirus. The inset image shows the expressing barrel cortical area identifiable by mCherry fluorescence. Note the majority of barrel cortical neurons nearby the viral injected site expressing mCherry. (b) Western blots of total and membrane surface-biotinylated recombinant Magneto2.0 and TRPV4, and endogenous GluA1 and GluA2 in barrel cortical neurons after 7-10 days of expression. Each lane loaded with 40 μg proteins. (c) Relative levels of membrane surface-biotinylated vs. total Magneto2.0 (Magneto2.0: 9.9±2.5%; GluA1: 212.2±18.7%, n=10, Z=2.803, p<0.01; GluA2: 260.3±14.2%; n=12, Z=2. 803, p<0.01) and TRPV4 (TRPV4: 293.2±25.4%; GluA1: 209.9±14.6%, n=11, Z=-2.401, p<0.05; GluA2: 276.2±24.3%, n=11, Z=-1.607, p=0.29) compared to GluA1 and GluA2. Asterisks indicate p<0.05 (Wilcoxon tests).

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