Journal: Molecular neurobiology

Article Title: Intracerebral expression of AAV-APOE4 is not sufficient to alter tau burden in two distinct models of tauopathy

doi: 10.1007/s12035-019-01859-4

Figure Lengend Snippet: Neonatal PS19 mice were injected with AAV-APOE4 or AAV-EGFP and aged to 3 months of age. APOE4 expression does not alter human tau transgene, as shown by CP27-immunoreactive total human tau (A). Quantification of tau protein band normalized to GAPDH loading control is shown (B). n=5 mice/group. Unpaired t test. Representative images of CP13 immunoreactivity from 3 month old mice are shown from different regions of the cortex and hippocampal CA1 (C). Scale bar: 100 μm. Quantitation of the immunostaining is presented as % immunoreactivity in the cortex and hippocampus (D). n= 5–11 mice/group. Two way ANOVA. Western blotting with PBS extracted brain lysate (denoted as S1) was done using ptau antibody (CP13, E-F) and total mouse tau antibody (T49, G-H) with protein band normalized to Gapdh loading control (F, H). n=5 mice/group. Unpaired t test. Pretangle tau levels were assessed by MC1 immunostaining in different areas of the cortex and hippocampus CA1 area (I) and data quantified as % immunoreactivity (J). Scale bar: 100 μm. n=5–11 mice/group. Two way ANOVA. Sarkosyl-insoluble cellular fraction (denoted as S4) was assessed for the presence of total human tau (K-L) and total mouse tau (M). n=5 mice/group. Unpaired t test. Black symbols, male mice; grey symbols, female mice.

Article Snippet: Statistical comparisons were conducted using unpaired t tests or 2 Way ANOVA with a Sidak post hoc test, if necessary (GraphPad Prism 7).

Techniques: Injection, Expressing, Control, Quantitation Assay, Immunostaining, Western Blot

Journal: Molecular neurobiology

Article Title: Intracerebral expression of AAV-APOE4 is not sufficient to alter tau burden in two distinct models of tauopathy

doi: 10.1007/s12035-019-01859-4

Figure Lengend Snippet: Morphometric analysis of the forebrain was done by counting hippocampal CA1–3 neurons and by measuring cortical thickness (frontal cortex, motor/sensory cortex, and auditory cortex) from specific brain areas of 4 month old rTg4510 mice (A-C) or 6 month old rTg4510 mice (J-L). This specific area was bounded by Bregma coordinates: 0.84mm and 1.08mm. Scale bar: 300 μm (larger panels on the left); 125μm (smaller panels on the right). n=4–6 mice/group. Unpaired t test of 2 way ANOVA. Synaptic protein profile was analyzed by immunoblotting for synaptogyrin 3, synaptophysin, spinophilin, PSD95, vGlut1 and GluR1 in the 4 month rTg4510 mice (D-I) and in the 6 month rTg4510 mice (M-R). Indicated protein bands were quantified following normalization to either GAPDH or Actin as shown. n=6/group (4 month rTg4510) and n=4 (6 month rTg4510). Black symbols, male mice; grey symbols, female mice.

Article Snippet: Statistical comparisons were conducted using unpaired t tests or 2 Way ANOVA with a Sidak post hoc test, if necessary (GraphPad Prism 7).

Techniques: Western Blot

Journal: Wellcome Open Research

Article Title: Single-transcript multiplex in situ hybridisation reveals unique patterns of dystrophin isoform expression in the developing mammalian embryo

doi: 10.12688/wellcomeopenres.15762.2

Figure Lengend Snippet: RNAscope probes to mouse dystrophin 5’ and 3’ regions recognise canine dystrophin in both healthy (WT) and dystrophic (deltaE50-MD) vastus lateralis muscle from 3-month-old ( A ) and 15-month-old ( C ) male animals. Nuclei (likely individual cells) associated with many 3’ probe foci are found within dystrophic muscle (arrowheads). Note dystrophin expression in blood vessel smooth muscle ( C , deltaE50-MD, lower panels). Apparent size distributions of probe foci ( B and D , upper panels) and subcellular localisations (lower panels) reveals 5’ foci fall into two populations: one small (~1.5 μm 2 ) and predominately sarcoplasmic; one large (>10 μm 2 ) and exclusively nuclear. 3’ foci are uniformly small (both sarcoplasmic and nuclear). DeltaE50-MD muscle shows no reduction in counts of sarcoplasmic 5’/3’ foci: instead nuclear counts of 3’ foci (commensurate with dp71 expression) tend to be higher. ( E ) qPCR for 5’ (exons 1-2) and 3’ (exons 62-64) regions of dp427m and for dp71 in 3- and 15-month-old WT and deltaE50-MD vastus lateralis muscles, confirms no significant dystrophy-associated reduction in dp427 transcriptional initiation and a modest (~2-fold) reduction in mature transcript number (P=0.006). A concomitant ~2-fold increase in dp71 expression (P=0.0005) is observed. All expression values are arbitrary units, normalised to HPRT1, RPL13a and SDHA (see methods) and adjusted relative to exon 1-2 values for comparative purposes. RNAscope ISH N=2-3 per genotype; qPCR N=6-7 per genotype, repeated measures 2-way ANOVA with Sidak’s multiple comparisons test (WT vs deltaE50-MD). Full-size figure can be found in the Underlying data .

Article Snippet: Statistical analysis of qPCR data ( vastus lateralis only) was performed with repeated measures 2-way ANOVA with Sidak’s multiple comparisons test (GraphPad Prism 8), with significance set at P<0.05.

Techniques: RNAscope, Expressing, Muscles