Journal: iScience

Article Title: Interleukin-6 promotes microtubule stability in axons via Stat3 protein–protein interactions

doi: 10.1016/j.isci.2021.103141

Figure Lengend Snippet: Axon transport deficits in IL-6−/− mice reflect a reduced rate of anterograde transport (A) Representative 40X images of dual-labeled βtubIII+/SMI-31+ RGCs from mid-central/mid-peripheral retina in WT (left) and IL-6−/− (right) mice. Quantification of RGC density showed no significant difference between WT and IL-6−/− mice (p = 0.284, n = 5–7 images/retina, 3 retina/genotype). Scale = 50 μm. Error bars = standard deviation. (B ) Representative images of ERRβ localization in the SC (left) and quantification of ERRβ signal density (right) suggests no alteration in the structure of RGC post-synaptic inputs to the SC between WT and IL-6−/− mice (p<0.05, n = 6 SC/genotype). Scale = 500 μm. Error bars = standard deviation. (C ) Representative 100X micrographs of CTB uptake (red) in WT (top) and IL-6−/− (bottom) retina immunolabeled with the RGC marker Brn3a (green). Inserts: Mander’s colocalization frequency. Scale = 50 μm. (D ) Top Panels: Localization of the caveolin-1 (Cav-1) to Brn3a+ RGC cell soma (red) in WT (left) and IL-6−/− (right) retina. Arrowheads indicate Cav-1+/Brn3a+ RGC soma. Scale = 20 μm. Bottom Panels: Immunoblotting (left) and densitometric quantification (right) of Cav-1 (22kDa) normalized to β-actin (42kDa) in retina from WT (white) and IL-6−/− (gray) mice. n = 3 retina/group. Error bars = standard deviation. (E) Left Panels: Representative coronal sections through the SC and respective retinotopic heat maps after 3 days of WGA transport in WT (left) and IL-6−/− mice (right). Arrows in coronal sections indicate areas of transport deficits. Density of the CTΒ signal for the heat maps range from 0% (blue) to 50% (green) to 70% (yellow) to 100% (red). Numbered dashed lines in retinotopic maps indicate the location of respective coronal section and white circles indicate position of the optic disk (OD). Medial (M) and rostral (R) orientations are indicated. Scale = 500 μm. Right Panel: Quantification of intact transport (≤70% Density). Dashed lines indicate median value and asterisks indicate p<0.01. n = 5–11 SC/genotype. Error bars = 95% confidence interval. (F ) Left Panels: Representative coronal sections through the SC and respective retinotopic heat maps after 5 days of CTB (left) or WGA (right) transport in IL-6−/− mice. OD= optic disk. Scale = 500 μm. Right Panel: Quantification of intact transport (≤70% Density). Dashed lines indicate median value. n = 5–11 SC/genotype. Error bars = 95% confidence interval.

Article Snippet: Porcine optic nerve (n = 12 from 7 individual pigs) was cut into small pieces with a scalpel and placed in IP solution containing: Pierce IP Lysis Buffer (10μl/mg tissue; Thermo Scientific, #87787), 1x Halt Protease and Phosphatase Inhibitor Cocktail (cat# 78440, Thermo Scientific), and 1x EDTA Solution (Thermo Scientific #78440, 100x stock of each).

Techniques: Labeling, Standard Deviation, Immunolabeling, Marker, Western Blot

Journal: iScience

Article Title: Interleukin-6 promotes microtubule stability in axons via Stat3 protein–protein interactions

doi: 10.1016/j.isci.2021.103141

Figure Lengend Snippet: IL-6 deficiency induces tubulin disorganization in RGC axons (A and B) Representative confocal micrographs (100X) of α-tubulin (A; red) and β-tubulin (βTubIII; B; green) immunolabeling in peripheral (left) and central (middle) regions of wholemount retina from WT (top) and IL-6−/− (bottom) mice. Arrowheads indicate regions with aggregated labeling. Boxed regions were zoomed to highlight aggregations (right). Scale = 20 μm. (C) Immunoblotting (top) and densitometric quantification (bottom) of α-tubulin (50 kDa) and β-tubulin (55 kDa) normalized to GAPDH (36 kDa) in retina and optic nerve from WT and IL-6−/− mice (n = 3). Error bars = standard deviation.

Article Snippet: Porcine optic nerve (n = 12 from 7 individual pigs) was cut into small pieces with a scalpel and placed in IP solution containing: Pierce IP Lysis Buffer (10μl/mg tissue; Thermo Scientific, #87787), 1x Halt Protease and Phosphatase Inhibitor Cocktail (cat# 78440, Thermo Scientific), and 1x EDTA Solution (Thermo Scientific #78440, 100x stock of each).

Techniques: Immunolabeling, Labeling, Western Blot, Standard Deviation

Journal: iScience

Article Title: Interleukin-6 promotes microtubule stability in axons via Stat3 protein–protein interactions

doi: 10.1016/j.isci.2021.103141

Figure Lengend Snippet: IL-6 directly promotes STAT3 and stathmin interaction in RGCs (A ) Representative confocal micrographs (100X) of STAT3:STMN PLA (red puncta) with DAPI counterstain (blue) in purified, primary RGCs following 4-day treatment with vehicle, recombinant IL-6 (rIL-6; 1ng/mL and 10ng/mL) or 10ng/mL rIL-6 + neutralizing antibody (nAb). Scale = 20 μm. (B) 3D plots of thermal luts for STAT3:STMN PLA signal in each micrograph pictured in (A). Thermal scale is 0 (purple) to 50 (red). (C) Quantification of PLA signal present in RGC neurites only. PLA signal is represented as integrated density/mm 2 (y-axis) for each treatment condition. Asterisks indicate p<0.05. Error bars represent standard deviation.

Article Snippet: Porcine optic nerve (n = 12 from 7 individual pigs) was cut into small pieces with a scalpel and placed in IP solution containing: Pierce IP Lysis Buffer (10μl/mg tissue; Thermo Scientific, #87787), 1x Halt Protease and Phosphatase Inhibitor Cocktail (cat# 78440, Thermo Scientific), and 1x EDTA Solution (Thermo Scientific #78440, 100x stock of each).

Techniques: Purification, Recombinant, Standard Deviation

Journal: Cell reports

Article Title: SoxD genes are required for adult neural stem cell activation

doi: 10.1016/j.celrep.2022.110313

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: After mechanical dissociation and washes with DMEM F12 (Thermo Fisher) to stop the reaction and washes with Hank’s, the disaggregated cell suspension was plated into MW12 plates with basal media [DMEM F12 + 1X N2 supplement (100X; Thermo Fisher) + 1X B27 supplement (50X; Thermo Fisher)], 20 ng/ml EGF (100 ng/ μl; PeproTech) and 20 ng/ml FGF-2 (100 ng/ μl; PeproTech).

Techniques: Virus, shRNA, Recombinant, Transfection, Luciferase, Reporter Assay, Plasmid Preparation, Software