Journal: Neurobiology of disease

Article Title: Protective effects of NAMPT or MAPK inhibitors and NaR on Wallerian degeneration of mammalian axons

doi: 10.1016/j.nbd.2022.105808

Figure Lengend Snippet: Western blot and Degeneration Index analysis reveal protective effects of DLK inhibition in transected axons. A-B. Semi-quantitative Western blot analysis of distal axons at 4 and 8 h after transection and treatment with the DLK inhibitor GNE3511 or vehicle (DMSO). A shows representative immunoblot and B quantification of signals normalized to actin and with reference to uninjured/untreated axon samples. GNE3511 suppresses phosphorylation of JNK at 4 h, t 5 = 6.17, p = 0.0005. NMNAT2 is significantly degraded between 4 and 8 h post-axotomy (t 5 = 3.5, p = 0.016) in vehicle treated axons but not in the presence of GNE (t 5 = 2.8, p = 0.046). Similarly, GNE suppresses the injury associated degradation of SCG10 at both time points (t 5 = 3.06, p = 0.031; and t 5 = 4.02, p = 0.008 respectively), and the injury associated degradation of spectrin at 8 h (calculated as the index of the p150 fragment to total spectrin, p284; t 5 = 3.08, p = 0.03). Signals were analyzed by 2-way ANOVAs with Holm-šídák’s multiple comparisons. C. Axons were treated with GNE 2 h before or at the time of transection, or 2 and 4 h later, or with vehicle (DMSO) at the time of injury. Statistical analysis of DI was performed by two-way ANOVA for the effect of time (F 7,35 = 164.7, p < 0.0001), treatment (F 4,5 = 47.78, p = 0.0004) and their interactions (F 28,35 = 16.22, p < 0.0001) with Holm-šídák’s multiple comparisons. GNE treatment at any time point offers protection against fragmentation early after axotomy (12 h ) compared to vehicle treated axons (t 40 = 5.7–8.8, p < 0.0001). However, pre-treatment or treatment at the time of injury shows significantly less degeneration compared to delayed treatment at 24 h (t 40 = 2.71–5.8, p < 0.03–0.0001). D. Representative photomicrographs from C. Scale bar, 50 μm. Error bars indicate ±1 SEM.

Article Snippet: Membranes were air dried, rehydrated and then blocked with 50% Odyssey blocking solution-tris buffered saline (Li-Cor, Germany) and incubated overnight at 4 °C with the target antibodies: mouse alpha-spectrin (Millipore Cat# MAB1622, RRID:AB_11214057) mouse actin (Santa Cruz Biotechnology Cat# sc-47,778 HRP, RRID:AB_2714189), rabbit SCG10 (Proteintech Cat# 10586–1-AP, RRID:AB_2197283), rabbit p Thr183/Tyr185 -JNK (Cell Signaling Technology Cat# 9251, RRID:AB_33165).

Techniques: Western Blot, Inhibition, Phospho-proteomics

Journal: PLoS ONE

Article Title: Increased Stathmin1 Expression in the Dentate Gyrus of Mice Causes Abnormal Axonal Arborizations

doi: 10.1371/journal.pone.0008596

Figure Lengend Snippet: (A, B) Comparison of stathmin1 expression of wild-type mice to that of Adcyap −/− mice. (A) Expression level of stathmin1 mRNA in DG was measured by real-time RT-PCR and normalized to the expression of GAPDH. Data represents means±SEM of independent experiments (wild type n = 10, Adcyap −/− n = 10, ***P = 0.0000496 compared with wild type). (B) Expression of stathmin1 protein in the DG of wild type and Adcyap −/− mice measured by western blot analysis (upper panel). GAPDH was used as internal control (middle panel). Lower panel shows the ratio of stathmin1 protein level in DG of Adcyap −/− mice (closed column) to that of wild-type mice (open column). Error bars represent ±SEM. (wild-type n = 4, Adcyap −/− n = 4, **P = 0.00277 compared with wild-type). (C, D) Kinetic studies of the effect of PACAP signaling on stahmin1 expression levels in PC12 cells. (C) Alteration of stathmin1 mRNA levels by the indicated period of PACAP (100 nM) stimulation was quantified by real-time PCR. Data are expressed as mean percentages ±SEM relative to control values at 0 h. Open circle indicates vehicle treatment. Closed circle represents PACAP treatment. (D) Alteration of stathmin1 protein levels under the indicated period of PACAP (100 nM) stimulation was measured by western blot analysis using an anti-stathmin1 antibody.

Article Snippet: The following antibodies were employed: rabbit polyclonal anti-stathmin1 (GeneTex).

Techniques: Comparison, Expressing, Quantitative RT-PCR, Western Blot, Control, Real-time Polymerase Chain Reaction

Journal: PLoS ONE

Article Title: Increased Stathmin1 Expression in the Dentate Gyrus of Mice Causes Abnormal Axonal Arborizations

doi: 10.1371/journal.pone.0008596

Figure Lengend Snippet: (A) Alteration of stathmin1 mRNA levels 24 hours after PACAP treatment, at indicated concentrations, was quantified by real-time PCR. Data are expressed as mean percentages ±SEM relative to control values (n = 3, PACAP 1 nM **P = 0.0044, 10 nM **P = 0.0033, 100 nM ***P = 0.0003 compared with control). (B, C) Effect of PACAP signaling pathway inhibitors on the PACAP-induced down-regulation of stathmin1 expression. (B) PC12 cells were treated with 10 nM PACAP for 6 h and incubated with or without the indicated concentration of PAC 1 /VPAC 2 receptor antagonist, PACAP 6-38 (n = 3, PACAP6-38 5 µM **P = 0.0027, 10 µM ***P = 0.00045 compared with PACAP stimulation alone) and (C) pretreatment of either ERK or p38 inhibitor (n = 3, PACAP stimulation alone **P = 0.0012, SB202190 ***P = 0.0008, ##P = 0.005, PD98059 *P = 0.018, #P = 0.0018, SB202190 & PD98059 #P = 0.013, *compared with each control, #compared with PACAP stimulation alone). Then stathmin1 expression was quantified by real-time PCR. Data are expressed as mean ratios ±SEM relative to control values. (D) Alteration of stathmin1 mRNA levels in PC12 cells, 6 hours after PACAP or VIP treatment at the indicated concentrations, was quantified by real-time PCR. Data are expressed as mean percentages ±SEM relative to control values (n = 3, PACAP 100 nM ***P = 7.08E −07 compared with control).

Article Snippet: The following antibodies were employed: rabbit polyclonal anti-stathmin1 (GeneTex).

Techniques: Real-time Polymerase Chain Reaction, Control, Expressing, Incubation, Concentration Assay

Journal: PLoS ONE

Article Title: Increased Stathmin1 Expression in the Dentate Gyrus of Mice Causes Abnormal Axonal Arborizations

doi: 10.1371/journal.pone.0008596

Figure Lengend Snippet: (A–B) Immunohistochemical analysis of stathmin1 in the DG. Immunohistochemistry of stathmin1 using wild-type mice (A) and Adcyap −/− mice (B). The small photograph is at high magnification. Bars = 50 µm. (C) The number of stathmin1 positive cells in the SGZ in wild-type (open column) and Adcyap −/− mice (closed column) were counted in a set of four coronal sections along the rostral-caudal range of each hippocampus/DG (n = 3). (D–G) Immunoelectron microscopic analysis of stathmin1-expressing processes in the SGZ of Adcyap −/− mice. (D, E) Stathmin1 positive neurites originating from SGZ neurons extended to the polymorphic layer (D, bar = 5 µm) and the neurites had the morphology of axon fibers (E, bar = 1 µm). (F) Stathmin1 positive dot-like fibers in the polymorphic layer identified as the axon fibers. Bar = 1 µm (G) Thick processes found in the granular cell layer could often be traced to the stathmin1 positive cell soma. Bar = 5 µm (H, K) Localization of stathmin1 in the rat hippocampal primary neurons. The same cells as in (H) and (K) stained with MAP2 antibody (I) or anti-Tau antibody (L). (J) and (M) represent the merged image of (H) with (I), and (K) with (L), respectively. Bars = 50 µm.

Article Snippet: The following antibodies were employed: rabbit polyclonal anti-stathmin1 (GeneTex).

Techniques: Immunohistochemical staining, Immunohistochemistry, Expressing, Staining

Journal: PLoS ONE

Article Title: Increased Stathmin1 Expression in the Dentate Gyrus of Mice Causes Abnormal Axonal Arborizations

doi: 10.1371/journal.pone.0008596

Figure Lengend Snippet: (A, B) Immunohistochemical analysis of stathmin1 in the polymorphic layer. Immunohistochemistry of stathmin1 using wild-type mice (A) and Adcyap −/− mice (B). White arrows indicate varicosities. Bars = 50 µm. (C) The number of stathmin1 positive dot-like fibers in the polymorphic layer were counted (n = 3, ***P = 3.4E −12 compared with wild-type). Error bars represent ±SEM. (D–F) Morphology of hippocampal primary neurons transfected with stathmin1. Over-expression of GFP (D), GFP-stathmin1 (E) or stathmin1 and GFP (F). The neurons over-expressing stathmin1 have abnormally pronounced sprouting of axon fibers. Bars = 50 µm. (G) The number of secondary neurites from axons was increased by over-expression of stathmin1 (GFP; n = 10, GFP-stathmin1; n = 10, ***P = 2.5E −5 , GFP and stathmin1; n = 6, ***P = 0.00022 compared with GFP). Error bars represent ±SEM.

Article Snippet: The following antibodies were employed: rabbit polyclonal anti-stathmin1 (GeneTex).

Techniques: Immunohistochemical staining, Immunohistochemistry, Transfection, Over Expression, Expressing

Journal: PLoS ONE

Article Title: Increased Stathmin1 Expression in the Dentate Gyrus of Mice Causes Abnormal Axonal Arborizations

doi: 10.1371/journal.pone.0008596

Figure Lengend Snippet: (A) Localization of stathmin1 and PAC 1 expressing neurons in the DG of wild-type mice. (B) Hippocampal section stained with hematoxylin and eosin. Bars = 100 µm. (C) shows the localization of stathmin1 and PAC 1 expressing neurons in the same section (arrows). Stathmin1 is visualized by immunohistochemistry (brown) and PAC 1 mRNA by in situ hybridization (black grains). Bars = 50 µm. (D–J) Involvement of Ascl1 in the promoter activity of stathmin1. (D) Schematic diagram of the 1.8 kb rat stathmin1 5′ genomic DNA fragments for the luciferase activity assay. These vectors posses the 5′ regulatory region of stathmin1, the first exon and partial sequence of the first intron. The different promoter constructs used in this study are shown. The relative location (not drawn to scale) of putative E-boxes, and the transcription start site (arrow at +1) are shown. (E) PC12 cells were transfected in 6-well plates with the indicated stathmin1 promoter-luciferase plasmids and the Renilla luciferase plasmid. Firefly and Renilla luciferase activities were measured using a Dual Luciferase Reporter Assay System (n = 3). Luciferase activity was measured and data (means±SEM) are shown as relative luciferase activity (RLA). (F) Activation of the STMN1-1 promoter by Ascl1. STMN1-1 plasmid and 1 µg of expression vector, which are mixed with empty vector pCI and the indicated amounts of pCI-Ascl1, were transfected in triplicate, with 0.2 µg. 48 h after PC12 cell transfection, the cells were collected and the RLA was determined. Data (mean±SEM) show representative results of least three experiments, *P = 0.0165. (G) Sheared chromatin from PC12 cells was immunoprecipitated with the appropriate antibody (anti-Ascl1, control rat anti-IgG). Then DNA sequences bound to precipitated proteins were isolated. Immunoprecipitated DNA was used as template in PCR using E10 primers, E10–E11 primers or negative control primers. (H–J) Localization of stathmin1 (H) and Ascl1 (I) were demonstrated on the same DG section of wild-type mice. Arrows indicate SGZ neurons containing both stathmin1 and Ascl1. Arrow head represents a neuron expressing stathmin1 alone. Bar = 10µm.

Article Snippet: The following antibodies were employed: rabbit polyclonal anti-stathmin1 (GeneTex).

Techniques: Expressing, Staining, Immunohistochemistry, In Situ Hybridization, Activity Assay, Luciferase, Sequencing, Construct, Transfection, Plasmid Preparation, Reporter Assay, Activation Assay, Immunoprecipitation, Control, Isolation, Negative Control

Journal: PLoS ONE

Article Title: Increased Stathmin1 Expression in the Dentate Gyrus of Mice Causes Abnormal Axonal Arborizations

doi: 10.1371/journal.pone.0008596

Figure Lengend Snippet: (A, B) Kinetic study of the effect of PACAP on the expression of Id3 (A) and Hes1 (B) mRNA. PC12 cells were treated with 10 nM PACAP for the indicated times and Id3 and Hes1 expression was quantified by real-time PCR. Data are expressed as mean percentages ±SEM relative to control values at 0 h. Open circle indicates vehicle treatment. Closed circle represents PACAP treatment. (C, D) Up-regulation of Id3 expression was inhibited by treatment with a p38 inhibitor (SB202190) (n = 3, SB202190 *P = 0.0202, SB202190 & PD98059 **P = 0.0032 compared with control) (C), and expression of Hes1 mRNA was inhibited by treatment with p38 (SB202190) and ERK (PD98059) inhibitors (n = 3, SB202190 ***P = 1.68×10 −5 , PD98059 *** P = 0.000252, SB202190 & PD98059 *** P = 8.98×10 −5 compared with control) (D). PC12 cells were incubated with each inhibitor for 1 h, then treated or not with 10 nM PACAP for 0.5 h. Id3 and Hes1 expression were quantified by real-time PCR. Data are expressed as mean ratios ±SEM relative to control values for inhibitor alone. (E) Effect of over-expression of Id3 or Hes1 on the promoter activity of stathmin1. PC12 cells were transfected with STMN1-1 promoter-luciferase plasmid alone or with Id3 expression plasmid, pCI-Id3 or Hes1 expression plasmid, pCI-Hes1. Co-transfection of Id3 or Hes1 expression plasmids inhibited the activity of STMN1-1 promoter plasmid (n = 3, +pCI- Id3 P = 0.0743, +pCI-Hes1 *P = 0.0305 compared with +control vector). (F) Id3, but not Hes1, inhibited the activity of STMN1-1 promoter increased by co-transfection of Ascl1 expression plasmid, pCI-Ascl1 (n = 3, +pCI-neo +Ascl1 **P = 0.00385, +pCI-Hes1 +Ascl1 *P = 0.025 compared with +pCI-neo, +pCI- Id3 +Ascl1 # P = 0.0454 compared with +pCI-neo +Ascl1). (G) The effect of PACAP on the expression of Ascl1 mRNA in PC12 cells. Ascl1 expression was quantified by real-time PCR (n = 3, *P = 0.048 compared with control). Data are expressed as mean percentages ±SEM relative to control. (H, I) Effects of down-regulation of Hes1 on the expression level of Ascl1 mRNA. The effect of siRNAs for Hes1 are shown in (H) (n = 3, siRNA1 *P = 0.0159, siRNA2 **P = 0.00636 compared with control). The expression level of Ascl1 was quantified by real-time PCR (n = 3, siRNA1 P = 0.076, siRNA2 *P = 0.0162 compared with control). Data are expressed as mean ratio ±SEM relative to control values.

Article Snippet: The following antibodies were employed: rabbit polyclonal anti-stathmin1 (GeneTex).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Control, Incubation, Over Expression, Activity Assay, Transfection, Luciferase, Plasmid Preparation, Cotransfection

Journal: PLoS ONE

Article Title: Increased Stathmin1 Expression in the Dentate Gyrus of Mice Causes Abnormal Axonal Arborizations

doi: 10.1371/journal.pone.0008596

Figure Lengend Snippet: Comparison of mRNA levels of stathmin1 in the postmortem brain of control subjects (open column) and schizophrenia subjects (closed column) (A), or bipolar disorder subjects (B). The relative expression levels of the gene, normalized to GAPDH or PGK1, are presented. The expression of stathmin1 normalized to GAPDH (Mann-Whitney test, p = 0.0082) and PGK1 (Mann-Whitney test, p = 0.0047) were significantly increased in the schizophrenia group, compared with the control group.

Article Snippet: The following antibodies were employed: rabbit polyclonal anti-stathmin1 (GeneTex).

Techniques: Comparison, Control, Expressing, MANN-WHITNEY

Journal: bioRxiv

Article Title: Vector analysis of steerable mechanical tension across nuclear lamina

doi: 10.1101/462275

Figure Lengend Snippet: (A) MCF-7 cells were treated with ADM, H 2 O 2 , and CHX separately. The samples were stained for both β-actin and α-tubulin. The cells were also stained with DAPI to localize the nuclei. The images were captured through confocal laser microscopy after immunofluorescence staining (n = 10). (B) Intracellular nanoparticle size distribution in MCF-7 cells separately treated with ADM, H 2 O 2 , and CHX (n = 10). (C) MCF-7 cell intracellular osmotic pressure was measured via osmometery after the cells were exposed to ADM, H 2 O 2 , and CHX. (**: 0.001 < p < 0.05, ns: p > 0.05, Tukey-b test, n = 10). (D) P-cofilin, cofilin, actin, p-stathmin, stathmin, and tubulin levels in MCF-7 cells that had been treated with ADM, H 2 O 2 , or CHX (n = 6). (E) In the H 2 O 2 group, the ΔpNOP could be divided into two parts by comparing the total intensity of the protein nanoparticles, generated from MF and MT depolymerization, respectively (versus , line 1, row 2). Scale bar, 10 μm. All error bars represent SEM.

Article Snippet: Antibodies were purchased from commercial sources: rabbit anti-lamin B1 antibody (13435S, Cell Signaling Technology, Temecula, USA), rabbit anti-β-actin antibody (4970P, Cell Signaling Technology, Temecula, USA), mouse anti-tubulin-α antibody (T5168, Boster, Wuhan, China), rabbit anti-vimentin antibody (5741S, Cell Signaling Technology, Temecula, USA), rabbit anti-GFP antibody (2956S, Cell Signaling Technology, Temecula, USA), mouse anti-phospho-cofilin (Ser24) (bs-10252R, Bioss, Woburn, USA), mouse anti-cofilin antibody (PB0035, Boster, Wuhan, China), rabbit anti-phospho-stathmin 1 (Ser38) (bs-3432R, Bioss, Woburn, USA), and mouse anti-stathmin 1 antibody (BA1417, Boster, Wuhan, China).

Techniques: Staining, Microscopy, Immunofluorescence, Generated

Journal: bioRxiv

Article Title: Promoting axon regeneration by enhancing the non-coding function of the injury-responsive coding gene Gpr151

doi: 10.1101/2021.02.19.431965

Figure Lengend Snippet: (a) Schematic structure of Gpr151 gene in wild type control (black) or targeted knockout (KO) mice (blue). Gpr151 gene was disrupted by the insertion of LacZ -cassette in the CDS region. (b) Schematic diagram of timeline illustrating the pre-conditioning injury and sample preparation. Sciatic nerves of wild type control or KO mice were injured by crushing the nerves with forceps and dissected at 3 days after injury. The sciatic nerves were prepared as cryosections and subjected to immunohistochemistry with anti-SCG10 and βIII tubulin antibodies. L4,5 DRGs were subjected to RT-qPCR analysis (ÄÄCt) or adult DRG neuron cultures. (c) RT- qPCR analysis of mouse L4,5 DRGs. Average of normalized levels of wild type control and KO mRNA levels at 3 days after introducing sciatic nerve crush injury. The averages of relative Gpr151 mRNA levels from uninjured KO, injured control and KO samples was compared to the average level from control uninjured samples (set to 1). (n=6 for each condition; *** p <0.001, ANOVA followed by Tukey test). (d) Representative longitudinal sections of immunohistochemistry of sciatic nerves from in vivo axon regeneration assay. The longitudinal cryosections were immunostained with anti-SCG10 (white; top and yellow converted; bottom), a marker protein specifically labelling regenerating axon. βIII tubulin antibody was used for counterstaining to visualize the nerve sections (blue; bottom). The red dotted arrow line indicates the crush site. Scale bar, 500 μm. (e) Average of normalized SCG10 intensity from immunostained sections in (d). Immunostained SCG10 fluorescence intensity was acquired by measurement windows with 100 pixel-width (equivalent to 87.7 μm) from crush sites to distal parts of nerves sections at an every 100-pixel distance from ImageJ . The acquired intensity from every measurement window was normalized to the intensity of the crush site (n=7 for control mice and n=5 for KO mice; * p <0.05 by t -test; mean ± SEM). (f) Regeneration index calculated from (e). Regeneration index was defined as a distance of an indicated SCG10 intensity. Average distance of normalized SCG10 intensity 0.5 or 0.2 was calculated from (e) and presented as regeneration index (mm). ( # p = 0.59, ## p = 0.05 by t -test). (g) Mouse L4,5 DRG tissues were dissected at 3 days with (+SNI) or without (-SNI) sciatic nerve crush injury (SNI). DRG neurons were plated and cultured to monitor pre-conditioning effect-induced neurite outgrowth. Scale bar, 100 μm. (h) Average of the longest axon length from (g) (three independent biological replicates; total 6 mice; total 304, 313, 291, 283 cells for each condition, -SNI Control, -SNI KO, +SNI Control, +SNI KO; *** p <0.001 by ANOVA followed by Tukey tests). (i) Cumulative frequency of the longest axon length from (g). (j) Percentage of neurons in three categories of the longest axon length.

Article Snippet: Anti-HA antibody (Abcam, ab9110; RRID: AB_307019), anti-GPR151 antibody (Aviva Systems Biology, OAAF06441), anti-α tubulin antibody (Santa Cruz Biotechnology, sc-53030; RRID: AB_2272440), anti-c-Jun antibody (Cell Signaling Technology, 9165; RRID: AB_2130165), anti-SCG10 antibody (Novus Biologicals, NBP1-49461; RRID: AB_10011569), anti-βIII tubulin antibody (TUJ1, Abcam, ab41489, RRID: AB_727049), anti-GAPDH antibody (Santa Cruz Biotechnology, sc-32233; RRID: AB_627679), anti-FLAG antibody (Cell Signaling Technology, 14793; RRID: AB_2572291).

Techniques: Control, Knock-Out, Sample Prep, Immunohistochemistry, Quantitative RT-PCR, In Vivo, Marker, Fluorescence, Cell Culture

Journal: bioRxiv

Article Title: Promoting axon regeneration by enhancing the non-coding function of the injury-responsive coding gene Gpr151

doi: 10.1101/2021.02.19.431965

Figure Lengend Snippet: (a) Schematic diagram of AAV-control and AAV-5’UTRm adeno-associated virus vector and experimental timeline of in vivo axon regeneration assay in sciatic nerves (wk, weeks). U6, U6 promoter; Scrambled shRNA sequence 48-mer; T, transcription termination sequence; CMV, human cytomegalovirus immediate early enhancer/promoter; GFP, enhanced green fluorescence protein CDS. (b) Representative image of adult DRG neurons. L4,5 DRGs were dissected from AAV-injected mice and cultured for 12 hours and immunostained with βIII tubulin antibody. GFP fluorescence signal indicated AAV-mediated in vivo gene delivery and expression of target genes. Scale bar, 100 μm. (c) In vivo axon regeneration assay from crushed sciatic nerves. Representative longitudinal sections of sciatic nerves from AAV-control or AAV-5’UTRm- injected mice. Red dotted arrows indicate the injury site. Scale bar, 500μm. (d) Normalized SCG10 intensity measured from SCG10 immunostained sections in (c). Measurement window (100-pixel width, equivalent to 87.7 μm) was placed at the crush site and intensities were acquired at every 100-pixel to the distal part of the sections. (e) Regeneration index calculated from (d). (n=8 for AAV-control, 10 for AAV-5’UTRm; ** p <0.01, **** p <0.0001 by t -test; mean±SEM). (f) Experimental timeline of in vivo axon regeneration assay in optic nerves (wk, weeks). (g) Representative longitudinal sections of optic nerves from AAV-control or AAV-5’UTRm-injected mice. Regenerating axons were labeled by Alexa594-conjugated CTB injection. Red dotted arrows indicate the injury site. Red bars indicated the locations analyzed the estimation of regenerating axon numbers in (h). Scale bar, 200μm. (h) Estimated numbers of regenerating axons (n=5 for AAV-control, 6 for AAV-5’UTRm; * p <0.05, ** p <0.01, *** p <0.001 by one-way AVONA with Bonferroni test; mean±SEM). (i) Relative expression level of Gpr151 and Atf3 after injury in mouse retina tissues (injured / uninjured). RT-qPCR analysis of Gpr151 and Atf3 from mouse retina tissues dissected at 3 days with optic nerve crush injury (injured, purple) or without injury (uninjured, black) (n=3 for each condition; *** p <0.001, ns, not significant by t-test; mean±SEM).

Article Snippet: Anti-HA antibody (Abcam, ab9110; RRID: AB_307019), anti-GPR151 antibody (Aviva Systems Biology, OAAF06441), anti-α tubulin antibody (Santa Cruz Biotechnology, sc-53030; RRID: AB_2272440), anti-c-Jun antibody (Cell Signaling Technology, 9165; RRID: AB_2130165), anti-SCG10 antibody (Novus Biologicals, NBP1-49461; RRID: AB_10011569), anti-βIII tubulin antibody (TUJ1, Abcam, ab41489, RRID: AB_727049), anti-GAPDH antibody (Santa Cruz Biotechnology, sc-32233; RRID: AB_627679), anti-FLAG antibody (Cell Signaling Technology, 14793; RRID: AB_2572291).

Techniques: Control, Virus, Plasmid Preparation, In Vivo, shRNA, Sequencing, Fluorescence, Injection, Cell Culture, Expressing, Labeling, Quantitative RT-PCR

Journal: eLife

Article Title: Translatome analysis reveals cellular network in DLK-dependent hippocampal glutamatergic neuron degeneration

doi: 10.7554/eLife.101173

Figure Lengend Snippet:

Article Snippet: STMN2 antibodies were a mouse monoclonal anti-STMN2 (R&D Systems, MAB6930) and a rabbit polyclonal anti-STMN2 (Proteintech, 10586–1-AP).

Techniques: Knock-Out, Over Expression, Sequencing, RNAscope, TUNEL Assay, SYBR Green Assay, Bicinchoninic Acid Protein Assay, Multiplex Assay, Software