Journal:

Article Title: THE RNA BINDING AND TRANSPORT PROTEINS STAUFEN AND FRAGILE X MENTAL RETARDATION PROTEIN ARE EXPRESSED BY RAT PRIMARY AFFERENT NEURONS AND LOCALIZE TO PERIPHERAL AND CENTRAL AXONS

doi: 10.1016/j.neuroscience.2006.05.047

Figure Lengend Snippet: Assessment of stau2 and FMRP protein and mRNA expression in the dorsal horn (DH), dorsal root (DR) and DRG by Western blot and RT-PCR. (A) Stau2 protein Western blot illustrating that the 62, 59 and 52 kDa isoforms of the protein are present in the DH and DRG, whereas only the 59 and 52 kDa isoforms are present in the DR. βIII-Tubulin (tub) was used as a loading control. (B) Stau1 mRNA was present in the DH, DR and DRG by RT-PCR, whereas stau2 mRNA was only present in the DH and DRG. GAPDH was used as a loading and positive control. (C) FMRP protein, detected as an ~80 kDa protein band, was present in DH, DR and DRG by Western blot. Cortex (Cor) protein was used as a positive control for the known size of FMRP. Tub was used as a loading control. (D) FMRP mRNA was detected by RT-PCR in the DH, DR and DRG. GAPDH was used as a loading and positive control. All Western blot and RT-PCR experiments are representative of samples run from three different animals per tissue.

Article Snippet: As a loading control, blots were then stripped using Restore reagent (Pierce, Rockford, IL, USA) and reblotted with a 1:1000 dilution of anti- β -III tubulin antibody (Tuj-1 clone, Promega).

Techniques: Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction, Positive Control

Journal: International Journal of Ophthalmology

Article Title: Promotion of axon regeneration and inhibition of astrocyte activation by alpha A-crystallin on crushed optic nerve

doi: 10.18240/ijo.2016.07.04

Figure Lengend Snippet: A: Immunofluorescence staining showed the GFAP-positive fibers (green) and TUJ1-positive processes (red) in the crush site following ONC. The sections were obtained from the ONC injury group (injury only) (a-d), PBS-treated ONC injury group (PBS injection) (e-h) and αA-crystallin-treated ONC injury group (α-A injection) (i-l). B, C: WB analysis showed that the GFAP levels in the optic nerve were significantly decreased in the αA-crystallin-treated group compared to the injury only (P<0.01) and PBS-treated groups (P<0.01). However, the TUJ1 levels were higher in the αA-crystallin-treated group compared to the injury only group (P<0.05). cr: The middle of crush site; ONC: Optic nerve crush. The quantitative data represent the means±SD (n=5). aP<0.05, bP<0.01. Scale bar=100 µm.

Article Snippet: The sections were then incubated with the following primary antibodies in 3% goat serum overnight at 4°C: rabbit anti-glial fibrillary acidic protein (GFAP) (Abcam, Cambrige, MA, USA, 1:500), mouse anti-beta Tubulin (TUJ1) monoclonal antibody (Earthox, San Francisco, USA, 1:300), mouse anti-feline CSPG Brain core protein (US Biological, US, 1:400), and mouse anti-neurocan antibody (EMD Millipore, 1:500).

Techniques: Immunofluorescence, Staining, Injection

Journal: Frontiers in Aging Neuroscience

Article Title: Age-Dependent Activation and Neuronal Differentiation of Lgr5+ Basal Cells in Injured Olfactory Epithelium via Notch Signaling Pathway

doi: 10.3389/fnagi.2020.602688

Figure Lengend Snippet: Aging altered neuronal generation in the olfactory epithelium (OE). (A) Immunostaining against mature and immature sensory neuronal markers olfactory marker protein (OMP) and TUJ1 in the OE of young and aged mice at 3- and 25-month old. The positive signals against OMP were labeled by dash lines. (B) Immunostaining against sensory neuronal marker PGP9.5 (noted by asterisks) and columnar-ciliated respiratory epithelial cell marker Krt19 (noted by dash lines). (C–L) Confocal images of staining against OMP and TUJ1 (C,D) , PGP9.5 (E,F) , Krt19 ( G,H , for respiratory epithelial cells, labeled by arrows), Sox2 ( I,J , for apical supporting cells), and Krt14 ( K,L , for HBCs) in the OE of young and aged mice. Nuclei were counterstained with DAPI, as shown in blue. (M) Statistical analysis of OMP+, TUJ1+, Krt19+, Krt14+, and apical Sox2+ cell density in the OE of young and aged mice. In young and aged OE, 1,406 and 723 OMP+, 262 and 179 TUJ1+, 162 and 133 Krt19+, 98 and 139 Krt14+, and 182 and 222 apical Sox2+ cells were counted. (N) Statistical analysis of OE thickness from young and aged mice. Each OE thickness was calculated from 18 sections of three mice. The statistical significance was determined by unpaired t -test with Mann–Whitney correction. ** p = 0.0072, *** p < 0.001 in (M) , * p = 0.3151 in (N) . Scale bars: 0.5 mm in (B) and 10 μm in (L) .

Article Snippet: The primary antibodies used in the immunostaining and their marked cellular subtypes, dilutions, vendors, and category numbers were as follows: rabbit anti-OMP (#ab87338, Abcam, 1:200, mature olfactory sensory neurons), mouse anti-TUJ1 (#ab78078, Abcan, 1:200, immature olfactory sensory neurons), rabbit anti-PGP9.5 (#14730-1-AP, Proteintech, 1:400, olfactory sensory neurons), mouse anti-Krt14 (#10143-1-AP, Proteintech, 1:200, horizontal basal cells), mouse anti-Krt19 (TROMA-III-c, DSHB, 1:300, ciliated respiratory epithelial cells), rabbit anti-Sox2 (#ab92494, Abcam, 1:200, supporting and basal cells), chicken anti-GFP (#ab13970, Abcam, 1:1,000, Lgr5-EGFP+ cells), rabbit anti-GFP (#A11122, Thermo Fisher Scientific, 1:500, Lgr5-EGFP+ cells), goat anti-Sox2 (#sc-17320, Santa Cruz Biotechnology, 1:200, supporting and basal cells), mouse anti-Ki67 (#550609, BD Biosciences, 1:100, proliferative basal cells), goat anti-ICAM1(#AF796, R&D, 1:500, horizontal basal cells), and mouse anti-Mash1 (#556604, BD Biosciences, 1:100, globose basal cells).

Techniques: Immunostaining, Marker, Labeling, Staining, MANN-WHITNEY