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    Feinstein Institute immersion mc1
    Cortical micrographs of transgenic rTg4510 (Tg) aged 12 (A-B; G-H; M-N), 16 (C-D; I-J; O-P) or 20 (E-F; K-L; Q-R) weeks-old, displaying the progression of tau pathology, showing no reactivity in control (Ctr) littermates (A, C, E) and age-related increase of tau immunoreactivity with <t>MC1</t> antibody starting at ~16 weeks (B, D, F). Cortical astrocyte reactivity and density also increased in Tg mice starting at ~12 weeks (H, J, L) in contrast to age-matched Ctr (G, I, K). Microglia reactivity and density in cortex of Tg mice also increased in an age-related manner in transgenic mice (N, P, R), in contrast to Ctr (M, O, Q). Scale bar 100 μm.
    Immersion Mc1, supplied by Feinstein Institute, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/immersion+mc1/bio_rxiv__500801-174-32-38?v=Feinstein+Institute
    Average 86 stars, based on 1 article reviews
    immersion mc1 - by Bioz Stars, 2026-06
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    Images

    1) Product Images from "Farnesyl Transferase Inhibition for the Treatment of Tauopathies"

    Article Title: Farnesyl Transferase Inhibition for the Treatment of Tauopathies

    Journal: bioRxiv

    doi: 10.1101/500801

    Cortical micrographs of transgenic rTg4510 (Tg) aged 12 (A-B; G-H; M-N), 16 (C-D; I-J; O-P) or 20 (E-F; K-L; Q-R) weeks-old, displaying the progression of tau pathology, showing no reactivity in control (Ctr) littermates (A, C, E) and age-related increase of tau immunoreactivity with MC1 antibody starting at ~16 weeks (B, D, F). Cortical astrocyte reactivity and density also increased in Tg mice starting at ~12 weeks (H, J, L) in contrast to age-matched Ctr (G, I, K). Microglia reactivity and density in cortex of Tg mice also increased in an age-related manner in transgenic mice (N, P, R), in contrast to Ctr (M, O, Q). Scale bar 100 μm.
    Figure Legend Snippet: Cortical micrographs of transgenic rTg4510 (Tg) aged 12 (A-B; G-H; M-N), 16 (C-D; I-J; O-P) or 20 (E-F; K-L; Q-R) weeks-old, displaying the progression of tau pathology, showing no reactivity in control (Ctr) littermates (A, C, E) and age-related increase of tau immunoreactivity with MC1 antibody starting at ~16 weeks (B, D, F). Cortical astrocyte reactivity and density also increased in Tg mice starting at ~12 weeks (H, J, L) in contrast to age-matched Ctr (G, I, K). Microglia reactivity and density in cortex of Tg mice also increased in an age-related manner in transgenic mice (N, P, R), in contrast to Ctr (M, O, Q). Scale bar 100 μm.

    Techniques Used: Transgenic Assay

    (A) Representative image of brain coronal section tiling 800 x 800 pixels Z-Stacks mosaic labeled with MC1 (Green), GFAP (Red) and Hoechst (cell nuclei, blue) is shown. (B) The coronal section area of the micrographs was computed. At five weeks, rTg4510 transgenic mice (Tg) did not differ in brain size from their non-transgenic littermates (p=0.942), however at 20 weeks, the area was significantly reduced in transgenic mice compared to both age-matched controls and younger transgenic (p=6.94×10 −4 ). (C) 20-week-old control littermates (Ctr) are non-reactive to MC1 staining; (D) whereas 20 week old transgenic mice have a high density of strongly labeled neurons in both cerebral cortex (CTX) and hippocampal formation (HPF). (E) Quantification of NFTs/mm 2 in CTX and HPF. Representative micrographs showing microglia (Iba1 + cells) in 20 week old Ctr (F) and Tg (G) mice, and (H) quantification of microglia/mm 2 density, revealing that microglia lacks an age-related decline in rTg4510 transgenic mice. At 20-weeks, in the Ctr CTX there were 178.45±66.19 microglia/mm 2 and in the Tg CTX there were 444.20±55.91 microglia/mm 2 (p=1.9×10 −3 ). In the Ctr HPF, there were 236.74±17.68 microglia/mm 2 and in the Tg HPF there were 470.99±41.97 microglia/mm 2 (p=5.11×10 −3 ). (I) Astrocytes labeled with GFAP antibody in non-transgenic (Ctr) and (J) 20 week-old transgenic (Tg) mice show cortical astrogliosis accompanying the MC1 immunoreactivity in the rTg4510 mouse model. Activated hypertrophied astrocytes are shown in the inset. (K) GFAP + percentage area in the coronal sections was quantified. The cortical GFAP signal (ANOVA p=9.14×10 −4 ) quantified in Tg at 20 weeks increased in comparison of age-matched Ctr (p=9.2×10 −3 ). The cortical GFAP signal also increased in 20-week-old transgenic (Tg) compared to five-week-old Tg (p=0.020). The hippocampal GFAP signal between Tg and Ctr mice at 5 and 20 weeks did not differ (ANOVA p=0.676). Statistics shown for post-hoc Tukey-HSD test at *p<0.05, **p<0.01, ***p<1×10 −3 . Scale bar scale 1 mm.
    Figure Legend Snippet: (A) Representative image of brain coronal section tiling 800 x 800 pixels Z-Stacks mosaic labeled with MC1 (Green), GFAP (Red) and Hoechst (cell nuclei, blue) is shown. (B) The coronal section area of the micrographs was computed. At five weeks, rTg4510 transgenic mice (Tg) did not differ in brain size from their non-transgenic littermates (p=0.942), however at 20 weeks, the area was significantly reduced in transgenic mice compared to both age-matched controls and younger transgenic (p=6.94×10 −4 ). (C) 20-week-old control littermates (Ctr) are non-reactive to MC1 staining; (D) whereas 20 week old transgenic mice have a high density of strongly labeled neurons in both cerebral cortex (CTX) and hippocampal formation (HPF). (E) Quantification of NFTs/mm 2 in CTX and HPF. Representative micrographs showing microglia (Iba1 + cells) in 20 week old Ctr (F) and Tg (G) mice, and (H) quantification of microglia/mm 2 density, revealing that microglia lacks an age-related decline in rTg4510 transgenic mice. At 20-weeks, in the Ctr CTX there were 178.45±66.19 microglia/mm 2 and in the Tg CTX there were 444.20±55.91 microglia/mm 2 (p=1.9×10 −3 ). In the Ctr HPF, there were 236.74±17.68 microglia/mm 2 and in the Tg HPF there were 470.99±41.97 microglia/mm 2 (p=5.11×10 −3 ). (I) Astrocytes labeled with GFAP antibody in non-transgenic (Ctr) and (J) 20 week-old transgenic (Tg) mice show cortical astrogliosis accompanying the MC1 immunoreactivity in the rTg4510 mouse model. Activated hypertrophied astrocytes are shown in the inset. (K) GFAP + percentage area in the coronal sections was quantified. The cortical GFAP signal (ANOVA p=9.14×10 −4 ) quantified in Tg at 20 weeks increased in comparison of age-matched Ctr (p=9.2×10 −3 ). The cortical GFAP signal also increased in 20-week-old transgenic (Tg) compared to five-week-old Tg (p=0.020). The hippocampal GFAP signal between Tg and Ctr mice at 5 and 20 weeks did not differ (ANOVA p=0.676). Statistics shown for post-hoc Tukey-HSD test at *p<0.05, **p<0.01, ***p<1×10 −3 . Scale bar scale 1 mm.

    Techniques Used: Labeling, Transgenic Assay, Staining

    (A) Brain coronal section area in 20-week old rTg4510 transgenic mice (Tg) that received chronic oral administration of Lonafarnib (L80) is larger than that of untreated (Unt) or vehicle alone (Veh) Tg mice. Untreated and vehicle-treated mice measured 40.43±0.83 mm 2 and 40.02±0.73 mm 2 , respectively. Coronal section areas from lonafarnib-treated Tg animals averaged 44.55±0.95 mm 2 (untreated p=5.6×10 −3 , vehicle-treated p=2.4×10 −3 ). (B-C) Reduction of the extent of MC1 immunoreactivity in lonafarnib (L80) treated transgenic mice compared to untreated mice (Unt). Scale bar 1 mm. (D-G) Detail of insets of panels B and C showing representative MC1 immunoreactivity on cortex (CTX) and hippocampus (HPF) of either untreated (Unt) or lonafarnib treated (L80) in 20 week-old Tg mice; (H) large-scale coronal section mosaics quantified for MC1 + /mm 2 indicates a significant reduction of tau pahology after lonafarnib treatment (L80) when compared to untreated animals (Unt) or animals treated with vehicle alone (Veh). (I-L) Density of microglia in the CTX and HPF of transgenic mice treated with lonafarnib (L80) or left untreated (Unt) is shown by Iba1 immunolabeling. Hippocampal microglial reactivity declined upon lonafarnib treatment. (M) Microglia quantification of coronal section mosaics in both CTX and HPF of transgenic mice treated accordingly. In the HPF (ANOVA p=0.040), lonafarnib-treated Tg mice at 20 weeks had 343.33±13.24 Iba1 + cells/mm 2 compared to vehicle treated animals (488.38±11.53 Iba1+ cells/mm 2 ) or untreated controls (526.95±12.02 Iba1 + cells/mm 2 , p=0.05) and represents a significant reduction in microgliosis. No statistically significant differences were observed in the CTX (ANOVA p=0.211) of lonafarnib-treated animals (442.51±10.73 Iba1 + cells/mm 2 ) when compared to vehicle (425.30±7.94 Iba1 + cells/mm 2 ) or untreated animals (515.45±11.03 Iba1 + cells/mm 2 ). (N-Q) Astrocytes immunostained with GFAP in CTX or HPF of untreated and lonafarnib treated Tg mice, and (R) quantification of GFAP signal in full coronal slices. Neither lonafarnib (L80) nor vehicle alone (Veh) altered astrocytes in 20 weeks-old Tg mice (cortex ANOVA p=0.411, hippocampus ANOVA p=0.111). Statistics shown for post-hoc Tukey-HSD test at *p<0.05, **p<0.01, ***p<1×10 −3 . (S) Lonafarnib treatment (L80) reduced sumoylated cells compared to rTg4510 mice (Tg; p=2.55×10 −3 ). (T) Lonafarnib reduced ubiquitin labeled cells compared to rTg4510 mice (Tg; p=0.05). Counts of double-labeled cells in the micrographs were used to compute Bayesian probabilities of any cell to be tau immunoreactive (MC1 + ) given sumo + (U) or ubiquitin + (V) immunoreactivity. Lonafarnib also decreased these probabilities of double-labeling, in comparison to age-matched rTg4510 mice (Tg). Statistics shown for post-hoc Tukey-HSD test at *p<0.05, **p<0.01, ***p<1×10 −3 .Scale bar 100 μm.
    Figure Legend Snippet: (A) Brain coronal section area in 20-week old rTg4510 transgenic mice (Tg) that received chronic oral administration of Lonafarnib (L80) is larger than that of untreated (Unt) or vehicle alone (Veh) Tg mice. Untreated and vehicle-treated mice measured 40.43±0.83 mm 2 and 40.02±0.73 mm 2 , respectively. Coronal section areas from lonafarnib-treated Tg animals averaged 44.55±0.95 mm 2 (untreated p=5.6×10 −3 , vehicle-treated p=2.4×10 −3 ). (B-C) Reduction of the extent of MC1 immunoreactivity in lonafarnib (L80) treated transgenic mice compared to untreated mice (Unt). Scale bar 1 mm. (D-G) Detail of insets of panels B and C showing representative MC1 immunoreactivity on cortex (CTX) and hippocampus (HPF) of either untreated (Unt) or lonafarnib treated (L80) in 20 week-old Tg mice; (H) large-scale coronal section mosaics quantified for MC1 + /mm 2 indicates a significant reduction of tau pahology after lonafarnib treatment (L80) when compared to untreated animals (Unt) or animals treated with vehicle alone (Veh). (I-L) Density of microglia in the CTX and HPF of transgenic mice treated with lonafarnib (L80) or left untreated (Unt) is shown by Iba1 immunolabeling. Hippocampal microglial reactivity declined upon lonafarnib treatment. (M) Microglia quantification of coronal section mosaics in both CTX and HPF of transgenic mice treated accordingly. In the HPF (ANOVA p=0.040), lonafarnib-treated Tg mice at 20 weeks had 343.33±13.24 Iba1 + cells/mm 2 compared to vehicle treated animals (488.38±11.53 Iba1+ cells/mm 2 ) or untreated controls (526.95±12.02 Iba1 + cells/mm 2 , p=0.05) and represents a significant reduction in microgliosis. No statistically significant differences were observed in the CTX (ANOVA p=0.211) of lonafarnib-treated animals (442.51±10.73 Iba1 + cells/mm 2 ) when compared to vehicle (425.30±7.94 Iba1 + cells/mm 2 ) or untreated animals (515.45±11.03 Iba1 + cells/mm 2 ). (N-Q) Astrocytes immunostained with GFAP in CTX or HPF of untreated and lonafarnib treated Tg mice, and (R) quantification of GFAP signal in full coronal slices. Neither lonafarnib (L80) nor vehicle alone (Veh) altered astrocytes in 20 weeks-old Tg mice (cortex ANOVA p=0.411, hippocampus ANOVA p=0.111). Statistics shown for post-hoc Tukey-HSD test at *p<0.05, **p<0.01, ***p<1×10 −3 . (S) Lonafarnib treatment (L80) reduced sumoylated cells compared to rTg4510 mice (Tg; p=2.55×10 −3 ). (T) Lonafarnib reduced ubiquitin labeled cells compared to rTg4510 mice (Tg; p=0.05). Counts of double-labeled cells in the micrographs were used to compute Bayesian probabilities of any cell to be tau immunoreactive (MC1 + ) given sumo + (U) or ubiquitin + (V) immunoreactivity. Lonafarnib also decreased these probabilities of double-labeling, in comparison to age-matched rTg4510 mice (Tg). Statistics shown for post-hoc Tukey-HSD test at *p<0.05, **p<0.01, ***p<1×10 −3 .Scale bar 100 μm.

    Techniques Used: Transgenic Assay, Immunolabeling, Labeling

    Cortical micrographs showing double immunostaining using anti-sumo and MC1, or anti-ubiquitin and MC1 were used to compute Bayesian probabilities of any cell to be (A) sumo + or (B) ubiquitin + given tau immunoreactivity (MC1 + ). Lonafarnib had no statistically significant effect on either of these probabilities, whereas Rhes-miR altered sumo double-labelling of MC1 + cells (sumo + given MC1 + ANOVA p=1.61×10 −3 , ubiquitin + given MC1 + ANOVA p=0.174). Statistics shown for post-hoc Tukey-HSD test at *p<0.05, **p<0.01.
    Figure Legend Snippet: Cortical micrographs showing double immunostaining using anti-sumo and MC1, or anti-ubiquitin and MC1 were used to compute Bayesian probabilities of any cell to be (A) sumo + or (B) ubiquitin + given tau immunoreactivity (MC1 + ). Lonafarnib had no statistically significant effect on either of these probabilities, whereas Rhes-miR altered sumo double-labelling of MC1 + cells (sumo + given MC1 + ANOVA p=1.61×10 −3 , ubiquitin + given MC1 + ANOVA p=0.174). Statistics shown for post-hoc Tukey-HSD test at *p<0.05, **p<0.01.

    Techniques Used: Double Immunostaining

    Ten-week old transgenic rTg4510 mice were intracranially infected with AAV 2/5 viral vectors containing either U6, Rhes-miR or Rhes-WT constructs and sacrificed at 20 weeks of age. (A-C) Full mosaic immunohistochemistry of MC1 immunoreactivity following Rhes suppression by Rhes-miR have fewer NFTs than transduction with U6 or Rhes. (D-F) Full-mosaic immunohistochemistry of Iba1 was also reduced in mice treated with Rhes-miR AAV. Scale bar 1 mm. Quantification of micrographs (N=3), indicates (G) significantly increased coronal section area, Rhes-miR (40.53±0.70 mm 2 ) compared to U6 (37.33±0.36 mm 2 ; p=5.0×10 −3 ) or Rhes injected transgenic mice (36.41±0.79 mm 2 ; p=3.98×10 −4 ). (H) reduced number of MC1 + cells/mm 2 both in cerebral cortex (CTX) and hippocampal formation (HPF). The cortical density of MC1 + cells/mm 2 in animals injected with Rhes-miR was 78.76±8.875 MC1 + cells/mm 2 compared to U6 (157.81±12.56 MC1 + cells/mm 2 ; p=1.7×10 −3 ) or Rhes overexpression (172.15±13.120 MC1+ cells/mm 2 ; p=2.86×10 −4 ). In the HPF, Rhes-miR injected mice had 49.15±7.01 MC1+ cells/mm 2 compared to U6 (86.09±9.28 MC1+ cells/mm 2 ; p=0.013) or Rhes overexpression (92.86±9.64 MC1 + cells/mm 2 ; p=3.3×10 −3 ). (I) Reduced number of microglia/mm2 in CTX and HPF in Rhes-miR injected mice. Rhes-miR injected transgenic mice were significantly reduced in the CTX (U6 control p= 5.85×10 −5 , Rhes overexpression, p=0.031) and in the HPF (U6 control p=9.48×10 −4 , Rhes over-expression, p=5.0×10 −3 ). (J) Astrocytic immunoreactivity: In CTX, Rhes-miR significantly reduced GFAP immunoreactivity compared to Rhes overexpression (p=6.9×10 −3 ), but not from U6 controls (p=0.357). In HPF, Rhes-miR significantly reduced GFAP immunostaining compared to U6 controls (p=0.024), but not when compared to Rhes overexpression (p=0.248); (K-M) Double immunostaining of CTX using MC1 and anti-sumo, or (O-Q) anti-ubiquitin in 20-week old rTg4510 mice and immunostained cell densities quantified as sumo + /mm 2 or ubiquitin + /mm 2 . (M) Rhes-miR reduced sumoylated cells compared to Rhes overexpression (p=5.4×10 −3 ). (Q) Rhes-miR injected mice did not show a significant reduction in ubiquitin labeled cells compared to either Rhes overexpression or U6 injected mice (p=0.615 or p=0.889, respectively). Counts of double-labeled cells in the micrographs were used to compute Bayesian probabilities of any cell to be tau immunoreactive (MC1 + ) given sumo + (N) or ubiquitin + (R) immunoreactivity. Rhes-miR treatments decreased these probabilities of double-labeling: (N) Rhes-miR (0.159±0.045 p=4.7×10 −3 ) significantly reduced the likelihood of sumoylated cells to double-label compared to Rhes overexpression. (R) Rhes-miR (0.0907±0.0112; p=2.5×10 −3 ) significantly reduced the likelihood of ubiquitinated cells to co-label compared to mice injected with U6. Statistics shown for post-hoc Tukey-HSD test at *p<0.05, **p<0.01, ***p<1×10 −3 . Scale bar 100 μm.
    Figure Legend Snippet: Ten-week old transgenic rTg4510 mice were intracranially infected with AAV 2/5 viral vectors containing either U6, Rhes-miR or Rhes-WT constructs and sacrificed at 20 weeks of age. (A-C) Full mosaic immunohistochemistry of MC1 immunoreactivity following Rhes suppression by Rhes-miR have fewer NFTs than transduction with U6 or Rhes. (D-F) Full-mosaic immunohistochemistry of Iba1 was also reduced in mice treated with Rhes-miR AAV. Scale bar 1 mm. Quantification of micrographs (N=3), indicates (G) significantly increased coronal section area, Rhes-miR (40.53±0.70 mm 2 ) compared to U6 (37.33±0.36 mm 2 ; p=5.0×10 −3 ) or Rhes injected transgenic mice (36.41±0.79 mm 2 ; p=3.98×10 −4 ). (H) reduced number of MC1 + cells/mm 2 both in cerebral cortex (CTX) and hippocampal formation (HPF). The cortical density of MC1 + cells/mm 2 in animals injected with Rhes-miR was 78.76±8.875 MC1 + cells/mm 2 compared to U6 (157.81±12.56 MC1 + cells/mm 2 ; p=1.7×10 −3 ) or Rhes overexpression (172.15±13.120 MC1+ cells/mm 2 ; p=2.86×10 −4 ). In the HPF, Rhes-miR injected mice had 49.15±7.01 MC1+ cells/mm 2 compared to U6 (86.09±9.28 MC1+ cells/mm 2 ; p=0.013) or Rhes overexpression (92.86±9.64 MC1 + cells/mm 2 ; p=3.3×10 −3 ). (I) Reduced number of microglia/mm2 in CTX and HPF in Rhes-miR injected mice. Rhes-miR injected transgenic mice were significantly reduced in the CTX (U6 control p= 5.85×10 −5 , Rhes overexpression, p=0.031) and in the HPF (U6 control p=9.48×10 −4 , Rhes over-expression, p=5.0×10 −3 ). (J) Astrocytic immunoreactivity: In CTX, Rhes-miR significantly reduced GFAP immunoreactivity compared to Rhes overexpression (p=6.9×10 −3 ), but not from U6 controls (p=0.357). In HPF, Rhes-miR significantly reduced GFAP immunostaining compared to U6 controls (p=0.024), but not when compared to Rhes overexpression (p=0.248); (K-M) Double immunostaining of CTX using MC1 and anti-sumo, or (O-Q) anti-ubiquitin in 20-week old rTg4510 mice and immunostained cell densities quantified as sumo + /mm 2 or ubiquitin + /mm 2 . (M) Rhes-miR reduced sumoylated cells compared to Rhes overexpression (p=5.4×10 −3 ). (Q) Rhes-miR injected mice did not show a significant reduction in ubiquitin labeled cells compared to either Rhes overexpression or U6 injected mice (p=0.615 or p=0.889, respectively). Counts of double-labeled cells in the micrographs were used to compute Bayesian probabilities of any cell to be tau immunoreactive (MC1 + ) given sumo + (N) or ubiquitin + (R) immunoreactivity. Rhes-miR treatments decreased these probabilities of double-labeling: (N) Rhes-miR (0.159±0.045 p=4.7×10 −3 ) significantly reduced the likelihood of sumoylated cells to double-label compared to Rhes overexpression. (R) Rhes-miR (0.0907±0.0112; p=2.5×10 −3 ) significantly reduced the likelihood of ubiquitinated cells to co-label compared to mice injected with U6. Statistics shown for post-hoc Tukey-HSD test at *p<0.05, **p<0.01, ***p<1×10 −3 . Scale bar 100 μm.

    Techniques Used: Transgenic Assay, Infection, Construct, Immunohistochemistry, Transduction, Injection, Over Expression, Immunostaining, Double Immunostaining, Labeling



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    immersion mc1  (Feinstein Institute)
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    Feinstein Institute immersion mc1
    Cortical micrographs of transgenic rTg4510 (Tg) aged 12 (A-B; G-H; M-N), 16 (C-D; I-J; O-P) or 20 (E-F; K-L; Q-R) weeks-old, displaying the progression of tau pathology, showing no reactivity in control (Ctr) littermates (A, C, E) and age-related increase of tau immunoreactivity with <t>MC1</t> antibody starting at ~16 weeks (B, D, F). Cortical astrocyte reactivity and density also increased in Tg mice starting at ~12 weeks (H, J, L) in contrast to age-matched Ctr (G, I, K). Microglia reactivity and density in cortex of Tg mice also increased in an age-related manner in transgenic mice (N, P, R), in contrast to Ctr (M, O, Q). Scale bar 100 μm.
    Immersion Mc1, supplied by Feinstein Institute, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/immersion+mc1/bio_rxiv__500801-174-32-38?v=Feinstein+Institute
    Average 86 stars, based on 1 article reviews
    immersion mc1 - by Bioz Stars, 2026-06
    86/100 stars
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    Image Search Results


    Cortical micrographs of transgenic rTg4510 (Tg) aged 12 (A-B; G-H; M-N), 16 (C-D; I-J; O-P) or 20 (E-F; K-L; Q-R) weeks-old, displaying the progression of tau pathology, showing no reactivity in control (Ctr) littermates (A, C, E) and age-related increase of tau immunoreactivity with MC1 antibody starting at ~16 weeks (B, D, F). Cortical astrocyte reactivity and density also increased in Tg mice starting at ~12 weeks (H, J, L) in contrast to age-matched Ctr (G, I, K). Microglia reactivity and density in cortex of Tg mice also increased in an age-related manner in transgenic mice (N, P, R), in contrast to Ctr (M, O, Q). Scale bar 100 μm.

    Journal: bioRxiv

    Article Title: Farnesyl Transferase Inhibition for the Treatment of Tauopathies

    doi: 10.1101/500801

    Figure Lengend Snippet: Cortical micrographs of transgenic rTg4510 (Tg) aged 12 (A-B; G-H; M-N), 16 (C-D; I-J; O-P) or 20 (E-F; K-L; Q-R) weeks-old, displaying the progression of tau pathology, showing no reactivity in control (Ctr) littermates (A, C, E) and age-related increase of tau immunoreactivity with MC1 antibody starting at ~16 weeks (B, D, F). Cortical astrocyte reactivity and density also increased in Tg mice starting at ~12 weeks (H, J, L) in contrast to age-matched Ctr (G, I, K). Microglia reactivity and density in cortex of Tg mice also increased in an age-related manner in transgenic mice (N, P, R), in contrast to Ctr (M, O, Q). Scale bar 100 μm.

    Article Snippet: Sections were immersed in normal donkey serum 1:20 in PBS containing 0.5% BSA, 0.1% Triton-X 100, and 0.1% sodium azide (PBTA) at 4°C on a rotator for continuous overnight agitation followed by immersion MC1 (provided by Peter Davies, Feinstein Institute for Medical Research, Manhasset, NY; mouse monoclonal, 1:200), anti-Iba1 (Wako laboratory chemicals, Richmond, VA; rabbit polyclonal, 1:200), anti-GFAP (abcam, San Francisco, CA; chicken polyclonal, 1:500), anti-ubiquitin (abcam, ab7780; rabbit polyclonal), and anti-sumo 1 (abcam, ab11672; rabbit monoclonal) diluted in PBTA.

    Techniques: Transgenic Assay

    (A) Representative image of brain coronal section tiling 800 x 800 pixels Z-Stacks mosaic labeled with MC1 (Green), GFAP (Red) and Hoechst (cell nuclei, blue) is shown. (B) The coronal section area of the micrographs was computed. At five weeks, rTg4510 transgenic mice (Tg) did not differ in brain size from their non-transgenic littermates (p=0.942), however at 20 weeks, the area was significantly reduced in transgenic mice compared to both age-matched controls and younger transgenic (p=6.94×10 −4 ). (C) 20-week-old control littermates (Ctr) are non-reactive to MC1 staining; (D) whereas 20 week old transgenic mice have a high density of strongly labeled neurons in both cerebral cortex (CTX) and hippocampal formation (HPF). (E) Quantification of NFTs/mm 2 in CTX and HPF. Representative micrographs showing microglia (Iba1 + cells) in 20 week old Ctr (F) and Tg (G) mice, and (H) quantification of microglia/mm 2 density, revealing that microglia lacks an age-related decline in rTg4510 transgenic mice. At 20-weeks, in the Ctr CTX there were 178.45±66.19 microglia/mm 2 and in the Tg CTX there were 444.20±55.91 microglia/mm 2 (p=1.9×10 −3 ). In the Ctr HPF, there were 236.74±17.68 microglia/mm 2 and in the Tg HPF there were 470.99±41.97 microglia/mm 2 (p=5.11×10 −3 ). (I) Astrocytes labeled with GFAP antibody in non-transgenic (Ctr) and (J) 20 week-old transgenic (Tg) mice show cortical astrogliosis accompanying the MC1 immunoreactivity in the rTg4510 mouse model. Activated hypertrophied astrocytes are shown in the inset. (K) GFAP + percentage area in the coronal sections was quantified. The cortical GFAP signal (ANOVA p=9.14×10 −4 ) quantified in Tg at 20 weeks increased in comparison of age-matched Ctr (p=9.2×10 −3 ). The cortical GFAP signal also increased in 20-week-old transgenic (Tg) compared to five-week-old Tg (p=0.020). The hippocampal GFAP signal between Tg and Ctr mice at 5 and 20 weeks did not differ (ANOVA p=0.676). Statistics shown for post-hoc Tukey-HSD test at *p<0.05, **p<0.01, ***p<1×10 −3 . Scale bar scale 1 mm.

    Journal: bioRxiv

    Article Title: Farnesyl Transferase Inhibition for the Treatment of Tauopathies

    doi: 10.1101/500801

    Figure Lengend Snippet: (A) Representative image of brain coronal section tiling 800 x 800 pixels Z-Stacks mosaic labeled with MC1 (Green), GFAP (Red) and Hoechst (cell nuclei, blue) is shown. (B) The coronal section area of the micrographs was computed. At five weeks, rTg4510 transgenic mice (Tg) did not differ in brain size from their non-transgenic littermates (p=0.942), however at 20 weeks, the area was significantly reduced in transgenic mice compared to both age-matched controls and younger transgenic (p=6.94×10 −4 ). (C) 20-week-old control littermates (Ctr) are non-reactive to MC1 staining; (D) whereas 20 week old transgenic mice have a high density of strongly labeled neurons in both cerebral cortex (CTX) and hippocampal formation (HPF). (E) Quantification of NFTs/mm 2 in CTX and HPF. Representative micrographs showing microglia (Iba1 + cells) in 20 week old Ctr (F) and Tg (G) mice, and (H) quantification of microglia/mm 2 density, revealing that microglia lacks an age-related decline in rTg4510 transgenic mice. At 20-weeks, in the Ctr CTX there were 178.45±66.19 microglia/mm 2 and in the Tg CTX there were 444.20±55.91 microglia/mm 2 (p=1.9×10 −3 ). In the Ctr HPF, there were 236.74±17.68 microglia/mm 2 and in the Tg HPF there were 470.99±41.97 microglia/mm 2 (p=5.11×10 −3 ). (I) Astrocytes labeled with GFAP antibody in non-transgenic (Ctr) and (J) 20 week-old transgenic (Tg) mice show cortical astrogliosis accompanying the MC1 immunoreactivity in the rTg4510 mouse model. Activated hypertrophied astrocytes are shown in the inset. (K) GFAP + percentage area in the coronal sections was quantified. The cortical GFAP signal (ANOVA p=9.14×10 −4 ) quantified in Tg at 20 weeks increased in comparison of age-matched Ctr (p=9.2×10 −3 ). The cortical GFAP signal also increased in 20-week-old transgenic (Tg) compared to five-week-old Tg (p=0.020). The hippocampal GFAP signal between Tg and Ctr mice at 5 and 20 weeks did not differ (ANOVA p=0.676). Statistics shown for post-hoc Tukey-HSD test at *p<0.05, **p<0.01, ***p<1×10 −3 . Scale bar scale 1 mm.

    Article Snippet: Sections were immersed in normal donkey serum 1:20 in PBS containing 0.5% BSA, 0.1% Triton-X 100, and 0.1% sodium azide (PBTA) at 4°C on a rotator for continuous overnight agitation followed by immersion MC1 (provided by Peter Davies, Feinstein Institute for Medical Research, Manhasset, NY; mouse monoclonal, 1:200), anti-Iba1 (Wako laboratory chemicals, Richmond, VA; rabbit polyclonal, 1:200), anti-GFAP (abcam, San Francisco, CA; chicken polyclonal, 1:500), anti-ubiquitin (abcam, ab7780; rabbit polyclonal), and anti-sumo 1 (abcam, ab11672; rabbit monoclonal) diluted in PBTA.

    Techniques: Labeling, Transgenic Assay, Staining

    (A) Brain coronal section area in 20-week old rTg4510 transgenic mice (Tg) that received chronic oral administration of Lonafarnib (L80) is larger than that of untreated (Unt) or vehicle alone (Veh) Tg mice. Untreated and vehicle-treated mice measured 40.43±0.83 mm 2 and 40.02±0.73 mm 2 , respectively. Coronal section areas from lonafarnib-treated Tg animals averaged 44.55±0.95 mm 2 (untreated p=5.6×10 −3 , vehicle-treated p=2.4×10 −3 ). (B-C) Reduction of the extent of MC1 immunoreactivity in lonafarnib (L80) treated transgenic mice compared to untreated mice (Unt). Scale bar 1 mm. (D-G) Detail of insets of panels B and C showing representative MC1 immunoreactivity on cortex (CTX) and hippocampus (HPF) of either untreated (Unt) or lonafarnib treated (L80) in 20 week-old Tg mice; (H) large-scale coronal section mosaics quantified for MC1 + /mm 2 indicates a significant reduction of tau pahology after lonafarnib treatment (L80) when compared to untreated animals (Unt) or animals treated with vehicle alone (Veh). (I-L) Density of microglia in the CTX and HPF of transgenic mice treated with lonafarnib (L80) or left untreated (Unt) is shown by Iba1 immunolabeling. Hippocampal microglial reactivity declined upon lonafarnib treatment. (M) Microglia quantification of coronal section mosaics in both CTX and HPF of transgenic mice treated accordingly. In the HPF (ANOVA p=0.040), lonafarnib-treated Tg mice at 20 weeks had 343.33±13.24 Iba1 + cells/mm 2 compared to vehicle treated animals (488.38±11.53 Iba1+ cells/mm 2 ) or untreated controls (526.95±12.02 Iba1 + cells/mm 2 , p=0.05) and represents a significant reduction in microgliosis. No statistically significant differences were observed in the CTX (ANOVA p=0.211) of lonafarnib-treated animals (442.51±10.73 Iba1 + cells/mm 2 ) when compared to vehicle (425.30±7.94 Iba1 + cells/mm 2 ) or untreated animals (515.45±11.03 Iba1 + cells/mm 2 ). (N-Q) Astrocytes immunostained with GFAP in CTX or HPF of untreated and lonafarnib treated Tg mice, and (R) quantification of GFAP signal in full coronal slices. Neither lonafarnib (L80) nor vehicle alone (Veh) altered astrocytes in 20 weeks-old Tg mice (cortex ANOVA p=0.411, hippocampus ANOVA p=0.111). Statistics shown for post-hoc Tukey-HSD test at *p<0.05, **p<0.01, ***p<1×10 −3 . (S) Lonafarnib treatment (L80) reduced sumoylated cells compared to rTg4510 mice (Tg; p=2.55×10 −3 ). (T) Lonafarnib reduced ubiquitin labeled cells compared to rTg4510 mice (Tg; p=0.05). Counts of double-labeled cells in the micrographs were used to compute Bayesian probabilities of any cell to be tau immunoreactive (MC1 + ) given sumo + (U) or ubiquitin + (V) immunoreactivity. Lonafarnib also decreased these probabilities of double-labeling, in comparison to age-matched rTg4510 mice (Tg). Statistics shown for post-hoc Tukey-HSD test at *p<0.05, **p<0.01, ***p<1×10 −3 .Scale bar 100 μm.

    Journal: bioRxiv

    Article Title: Farnesyl Transferase Inhibition for the Treatment of Tauopathies

    doi: 10.1101/500801

    Figure Lengend Snippet: (A) Brain coronal section area in 20-week old rTg4510 transgenic mice (Tg) that received chronic oral administration of Lonafarnib (L80) is larger than that of untreated (Unt) or vehicle alone (Veh) Tg mice. Untreated and vehicle-treated mice measured 40.43±0.83 mm 2 and 40.02±0.73 mm 2 , respectively. Coronal section areas from lonafarnib-treated Tg animals averaged 44.55±0.95 mm 2 (untreated p=5.6×10 −3 , vehicle-treated p=2.4×10 −3 ). (B-C) Reduction of the extent of MC1 immunoreactivity in lonafarnib (L80) treated transgenic mice compared to untreated mice (Unt). Scale bar 1 mm. (D-G) Detail of insets of panels B and C showing representative MC1 immunoreactivity on cortex (CTX) and hippocampus (HPF) of either untreated (Unt) or lonafarnib treated (L80) in 20 week-old Tg mice; (H) large-scale coronal section mosaics quantified for MC1 + /mm 2 indicates a significant reduction of tau pahology after lonafarnib treatment (L80) when compared to untreated animals (Unt) or animals treated with vehicle alone (Veh). (I-L) Density of microglia in the CTX and HPF of transgenic mice treated with lonafarnib (L80) or left untreated (Unt) is shown by Iba1 immunolabeling. Hippocampal microglial reactivity declined upon lonafarnib treatment. (M) Microglia quantification of coronal section mosaics in both CTX and HPF of transgenic mice treated accordingly. In the HPF (ANOVA p=0.040), lonafarnib-treated Tg mice at 20 weeks had 343.33±13.24 Iba1 + cells/mm 2 compared to vehicle treated animals (488.38±11.53 Iba1+ cells/mm 2 ) or untreated controls (526.95±12.02 Iba1 + cells/mm 2 , p=0.05) and represents a significant reduction in microgliosis. No statistically significant differences were observed in the CTX (ANOVA p=0.211) of lonafarnib-treated animals (442.51±10.73 Iba1 + cells/mm 2 ) when compared to vehicle (425.30±7.94 Iba1 + cells/mm 2 ) or untreated animals (515.45±11.03 Iba1 + cells/mm 2 ). (N-Q) Astrocytes immunostained with GFAP in CTX or HPF of untreated and lonafarnib treated Tg mice, and (R) quantification of GFAP signal in full coronal slices. Neither lonafarnib (L80) nor vehicle alone (Veh) altered astrocytes in 20 weeks-old Tg mice (cortex ANOVA p=0.411, hippocampus ANOVA p=0.111). Statistics shown for post-hoc Tukey-HSD test at *p<0.05, **p<0.01, ***p<1×10 −3 . (S) Lonafarnib treatment (L80) reduced sumoylated cells compared to rTg4510 mice (Tg; p=2.55×10 −3 ). (T) Lonafarnib reduced ubiquitin labeled cells compared to rTg4510 mice (Tg; p=0.05). Counts of double-labeled cells in the micrographs were used to compute Bayesian probabilities of any cell to be tau immunoreactive (MC1 + ) given sumo + (U) or ubiquitin + (V) immunoreactivity. Lonafarnib also decreased these probabilities of double-labeling, in comparison to age-matched rTg4510 mice (Tg). Statistics shown for post-hoc Tukey-HSD test at *p<0.05, **p<0.01, ***p<1×10 −3 .Scale bar 100 μm.

    Article Snippet: Sections were immersed in normal donkey serum 1:20 in PBS containing 0.5% BSA, 0.1% Triton-X 100, and 0.1% sodium azide (PBTA) at 4°C on a rotator for continuous overnight agitation followed by immersion MC1 (provided by Peter Davies, Feinstein Institute for Medical Research, Manhasset, NY; mouse monoclonal, 1:200), anti-Iba1 (Wako laboratory chemicals, Richmond, VA; rabbit polyclonal, 1:200), anti-GFAP (abcam, San Francisco, CA; chicken polyclonal, 1:500), anti-ubiquitin (abcam, ab7780; rabbit polyclonal), and anti-sumo 1 (abcam, ab11672; rabbit monoclonal) diluted in PBTA.

    Techniques: Transgenic Assay, Immunolabeling, Labeling

    Cortical micrographs showing double immunostaining using anti-sumo and MC1, or anti-ubiquitin and MC1 were used to compute Bayesian probabilities of any cell to be (A) sumo + or (B) ubiquitin + given tau immunoreactivity (MC1 + ). Lonafarnib had no statistically significant effect on either of these probabilities, whereas Rhes-miR altered sumo double-labelling of MC1 + cells (sumo + given MC1 + ANOVA p=1.61×10 −3 , ubiquitin + given MC1 + ANOVA p=0.174). Statistics shown for post-hoc Tukey-HSD test at *p<0.05, **p<0.01.

    Journal: bioRxiv

    Article Title: Farnesyl Transferase Inhibition for the Treatment of Tauopathies

    doi: 10.1101/500801

    Figure Lengend Snippet: Cortical micrographs showing double immunostaining using anti-sumo and MC1, or anti-ubiquitin and MC1 were used to compute Bayesian probabilities of any cell to be (A) sumo + or (B) ubiquitin + given tau immunoreactivity (MC1 + ). Lonafarnib had no statistically significant effect on either of these probabilities, whereas Rhes-miR altered sumo double-labelling of MC1 + cells (sumo + given MC1 + ANOVA p=1.61×10 −3 , ubiquitin + given MC1 + ANOVA p=0.174). Statistics shown for post-hoc Tukey-HSD test at *p<0.05, **p<0.01.

    Article Snippet: Sections were immersed in normal donkey serum 1:20 in PBS containing 0.5% BSA, 0.1% Triton-X 100, and 0.1% sodium azide (PBTA) at 4°C on a rotator for continuous overnight agitation followed by immersion MC1 (provided by Peter Davies, Feinstein Institute for Medical Research, Manhasset, NY; mouse monoclonal, 1:200), anti-Iba1 (Wako laboratory chemicals, Richmond, VA; rabbit polyclonal, 1:200), anti-GFAP (abcam, San Francisco, CA; chicken polyclonal, 1:500), anti-ubiquitin (abcam, ab7780; rabbit polyclonal), and anti-sumo 1 (abcam, ab11672; rabbit monoclonal) diluted in PBTA.

    Techniques: Double Immunostaining

    Ten-week old transgenic rTg4510 mice were intracranially infected with AAV 2/5 viral vectors containing either U6, Rhes-miR or Rhes-WT constructs and sacrificed at 20 weeks of age. (A-C) Full mosaic immunohistochemistry of MC1 immunoreactivity following Rhes suppression by Rhes-miR have fewer NFTs than transduction with U6 or Rhes. (D-F) Full-mosaic immunohistochemistry of Iba1 was also reduced in mice treated with Rhes-miR AAV. Scale bar 1 mm. Quantification of micrographs (N=3), indicates (G) significantly increased coronal section area, Rhes-miR (40.53±0.70 mm 2 ) compared to U6 (37.33±0.36 mm 2 ; p=5.0×10 −3 ) or Rhes injected transgenic mice (36.41±0.79 mm 2 ; p=3.98×10 −4 ). (H) reduced number of MC1 + cells/mm 2 both in cerebral cortex (CTX) and hippocampal formation (HPF). The cortical density of MC1 + cells/mm 2 in animals injected with Rhes-miR was 78.76±8.875 MC1 + cells/mm 2 compared to U6 (157.81±12.56 MC1 + cells/mm 2 ; p=1.7×10 −3 ) or Rhes overexpression (172.15±13.120 MC1+ cells/mm 2 ; p=2.86×10 −4 ). In the HPF, Rhes-miR injected mice had 49.15±7.01 MC1+ cells/mm 2 compared to U6 (86.09±9.28 MC1+ cells/mm 2 ; p=0.013) or Rhes overexpression (92.86±9.64 MC1 + cells/mm 2 ; p=3.3×10 −3 ). (I) Reduced number of microglia/mm2 in CTX and HPF in Rhes-miR injected mice. Rhes-miR injected transgenic mice were significantly reduced in the CTX (U6 control p= 5.85×10 −5 , Rhes overexpression, p=0.031) and in the HPF (U6 control p=9.48×10 −4 , Rhes over-expression, p=5.0×10 −3 ). (J) Astrocytic immunoreactivity: In CTX, Rhes-miR significantly reduced GFAP immunoreactivity compared to Rhes overexpression (p=6.9×10 −3 ), but not from U6 controls (p=0.357). In HPF, Rhes-miR significantly reduced GFAP immunostaining compared to U6 controls (p=0.024), but not when compared to Rhes overexpression (p=0.248); (K-M) Double immunostaining of CTX using MC1 and anti-sumo, or (O-Q) anti-ubiquitin in 20-week old rTg4510 mice and immunostained cell densities quantified as sumo + /mm 2 or ubiquitin + /mm 2 . (M) Rhes-miR reduced sumoylated cells compared to Rhes overexpression (p=5.4×10 −3 ). (Q) Rhes-miR injected mice did not show a significant reduction in ubiquitin labeled cells compared to either Rhes overexpression or U6 injected mice (p=0.615 or p=0.889, respectively). Counts of double-labeled cells in the micrographs were used to compute Bayesian probabilities of any cell to be tau immunoreactive (MC1 + ) given sumo + (N) or ubiquitin + (R) immunoreactivity. Rhes-miR treatments decreased these probabilities of double-labeling: (N) Rhes-miR (0.159±0.045 p=4.7×10 −3 ) significantly reduced the likelihood of sumoylated cells to double-label compared to Rhes overexpression. (R) Rhes-miR (0.0907±0.0112; p=2.5×10 −3 ) significantly reduced the likelihood of ubiquitinated cells to co-label compared to mice injected with U6. Statistics shown for post-hoc Tukey-HSD test at *p<0.05, **p<0.01, ***p<1×10 −3 . Scale bar 100 μm.

    Journal: bioRxiv

    Article Title: Farnesyl Transferase Inhibition for the Treatment of Tauopathies

    doi: 10.1101/500801

    Figure Lengend Snippet: Ten-week old transgenic rTg4510 mice were intracranially infected with AAV 2/5 viral vectors containing either U6, Rhes-miR or Rhes-WT constructs and sacrificed at 20 weeks of age. (A-C) Full mosaic immunohistochemistry of MC1 immunoreactivity following Rhes suppression by Rhes-miR have fewer NFTs than transduction with U6 or Rhes. (D-F) Full-mosaic immunohistochemistry of Iba1 was also reduced in mice treated with Rhes-miR AAV. Scale bar 1 mm. Quantification of micrographs (N=3), indicates (G) significantly increased coronal section area, Rhes-miR (40.53±0.70 mm 2 ) compared to U6 (37.33±0.36 mm 2 ; p=5.0×10 −3 ) or Rhes injected transgenic mice (36.41±0.79 mm 2 ; p=3.98×10 −4 ). (H) reduced number of MC1 + cells/mm 2 both in cerebral cortex (CTX) and hippocampal formation (HPF). The cortical density of MC1 + cells/mm 2 in animals injected with Rhes-miR was 78.76±8.875 MC1 + cells/mm 2 compared to U6 (157.81±12.56 MC1 + cells/mm 2 ; p=1.7×10 −3 ) or Rhes overexpression (172.15±13.120 MC1+ cells/mm 2 ; p=2.86×10 −4 ). In the HPF, Rhes-miR injected mice had 49.15±7.01 MC1+ cells/mm 2 compared to U6 (86.09±9.28 MC1+ cells/mm 2 ; p=0.013) or Rhes overexpression (92.86±9.64 MC1 + cells/mm 2 ; p=3.3×10 −3 ). (I) Reduced number of microglia/mm2 in CTX and HPF in Rhes-miR injected mice. Rhes-miR injected transgenic mice were significantly reduced in the CTX (U6 control p= 5.85×10 −5 , Rhes overexpression, p=0.031) and in the HPF (U6 control p=9.48×10 −4 , Rhes over-expression, p=5.0×10 −3 ). (J) Astrocytic immunoreactivity: In CTX, Rhes-miR significantly reduced GFAP immunoreactivity compared to Rhes overexpression (p=6.9×10 −3 ), but not from U6 controls (p=0.357). In HPF, Rhes-miR significantly reduced GFAP immunostaining compared to U6 controls (p=0.024), but not when compared to Rhes overexpression (p=0.248); (K-M) Double immunostaining of CTX using MC1 and anti-sumo, or (O-Q) anti-ubiquitin in 20-week old rTg4510 mice and immunostained cell densities quantified as sumo + /mm 2 or ubiquitin + /mm 2 . (M) Rhes-miR reduced sumoylated cells compared to Rhes overexpression (p=5.4×10 −3 ). (Q) Rhes-miR injected mice did not show a significant reduction in ubiquitin labeled cells compared to either Rhes overexpression or U6 injected mice (p=0.615 or p=0.889, respectively). Counts of double-labeled cells in the micrographs were used to compute Bayesian probabilities of any cell to be tau immunoreactive (MC1 + ) given sumo + (N) or ubiquitin + (R) immunoreactivity. Rhes-miR treatments decreased these probabilities of double-labeling: (N) Rhes-miR (0.159±0.045 p=4.7×10 −3 ) significantly reduced the likelihood of sumoylated cells to double-label compared to Rhes overexpression. (R) Rhes-miR (0.0907±0.0112; p=2.5×10 −3 ) significantly reduced the likelihood of ubiquitinated cells to co-label compared to mice injected with U6. Statistics shown for post-hoc Tukey-HSD test at *p<0.05, **p<0.01, ***p<1×10 −3 . Scale bar 100 μm.

    Article Snippet: Sections were immersed in normal donkey serum 1:20 in PBS containing 0.5% BSA, 0.1% Triton-X 100, and 0.1% sodium azide (PBTA) at 4°C on a rotator for continuous overnight agitation followed by immersion MC1 (provided by Peter Davies, Feinstein Institute for Medical Research, Manhasset, NY; mouse monoclonal, 1:200), anti-Iba1 (Wako laboratory chemicals, Richmond, VA; rabbit polyclonal, 1:200), anti-GFAP (abcam, San Francisco, CA; chicken polyclonal, 1:500), anti-ubiquitin (abcam, ab7780; rabbit polyclonal), and anti-sumo 1 (abcam, ab11672; rabbit monoclonal) diluted in PBTA.

    Techniques: Transgenic Assay, Infection, Construct, Immunohistochemistry, Transduction, Injection, Over Expression, Immunostaining, Double Immunostaining, Labeling