mouse α tuj1  (R&D Systems)

Journal: bioRxiv

Article Title: Activation of pro-survival autophagy by a small molecule promoting p62 oligomerisation

doi: 10.1101/2025.05.27.656309

Figure Lengend Snippet: A, Fluorescence microscopy images and quantification of mitophagy in Npc1 +/+ and Npc1 -/- MEFs expressing mt-mKeima cultured for 24 h in galactose medium supplemented with 50 nM rapamycin or 10 µM SQ-1. B , Fluorescence microscopy images and quantification of MitoSOX staining of Npc1 +/+ and Npc1 -/- MEFs cultured for 24 h in galactose medium supplemented with 50 nM rapamycin or 10 µM SQ-1. C , Phase-contrast images and cytotoxicity assay in Npc1 +/+ and Npc1 -/- MEFs cultured in galactose medium supplemented with 50 nM rapamycin or 10 µM SQ-1 for 72 h (phase contrast image) or 144 h (cytotoxicity assay). D , Cytotoxicity assay in control and NPC1 patient iPSC-derived cortical neurons after 4 weeks of neuronal differentiation, where neurons were treated with 50 nM rapamycin or 1-100 µM SQ-1 for 6 days. E , Fluorescence images and quantification of TUNEL + apoptotic nuclei in TUJ1 + cells treated as in D. F , Fluorescence images and quantification of LC3B and p62 puncta in TUJ1 + treated as in D. G , Schematic representation of the proposed mechanism by which SQ-1 sensitises p62 to mitochondria-generated ROS, leading to p62 oligomer formation, selective autophagy/mitophagy rescue, reduction in mitochondria with elevated ROS levels, and rescue of cell death. Data are mean ± SEM of n = 3 biological replicates. P values were calculated by one-way ANOVA followed by multiple comparisons with Dunnett’s test (A) or two-way ANOVA followed by multiple comparisons with Dunnett’s test (B, C, D). *P< 0.05; **P<0.01; ***P<0.001; ns (non-significant).

Article Snippet: The following primary antibodies were used: rabbit α-LC3B (NB100-2220, Novus Biologicals, 1:200), mouse α-p62 (610832, BD Biosciences, 1:200), mouse α-TUJ1 (4466, CST, 1:200).

Techniques: Fluorescence, Microscopy, Expressing, Cell Culture, Staining, Cytotoxicity Assay, Control, Derivative Assay, TUNEL Assay, Generated

Journal: eNeuro

Article Title: Oral Sensory Neurons of the Geniculate Ganglion That Express Tyrosine Hydroxylase Comprise a Subpopulation That Contacts Type II and Type III Taste Bud Cells

doi: 10.1523/ENEURO.0523-21.2022

Figure Lengend Snippet: TH expression in the GG. A , Fluorescence in situ hybridization labeling of GG sections from adult C57Bl/6 mice using probes to Phox2b (green) and Th (blue) revealed neurons that express high levels of Th (yellow arrow) and low levels of Th (white arrow). B , Immunofluorescence labeling of adult GG from Th -CreER; Rosa RFP mice using antibodies to TUJ1 (green), PHOX2B (blue), and RFP (magenta) identifies a population of PHOX2B+ neurons that also express RFP ( Th ). C , Immunofluorescence labeling of Th -WT; Rosa RFP mice, as in B , confirms the specificity of the RFP immunoreactions. Immunofluorescence labeling of nodose/petrosal/jugular ganglion complexes from Th -CreER; Rosa RFP mice using the same antibodies in B demonstrated the presence of Th +/PHOX2B+ oral sensory neurons in petrosal ganglia as well, shown in Extended Data . D , Serial sections of GG evaluated with both methods were counted and quantified to determine what percentage of Phox2b + neurons coexpressed Th . Only high Th -expressing neurons are graphed in D for the in situ hybridization. The p value is p = 0.0027. E , Serial sections of GG were quantified to determine the number of Th -expressing neurons that did not express Phox2b . Error bars represent mean ± SEM, and n = 3–4 mice were analyzed by each method. The p value is p = 0.2797. Scale bar in A is 100 μm, and the scale bar in B is 100 μm and applies to all panels in C . ** denotes p < 0.01, and ns is not statistically significant.

Article Snippet: The slides were then incubated with the following primary antibodies, which were diluted in PBS containing 0.3% Triton X-100 with 1% BSA in a humidified chamber overnight, or for 5 d (free floating tissues) at 4°C: rat α–cytokeratin-8 (TROMA-I, supernatant; 1:100 dilution, DSHB], rabbit α-RFP (1:200 dilution; Rockland, catalog #600-401-379), chicken α-RFP (1:200 dilution; Rockland, catalog #600-401-901), goat α-PHOX2B (1:200 dilution; R&D Systems, catalog #AF4940,), mouse α-TUJ1 (βIII-tubulin; 1:200 dilution; Sigma, catalog #T8578), goat α-CAR4 (1:200 dilution; R&D Systems, catalog #AF2414), and rabbit α-TRPM5 (1:200 dilution; Emily Liman’s Lab, University of Southern California).

Techniques: Expressing, Fluorescence, In Situ Hybridization, Labeling, Immunofluorescence

Journal: eNeuro

Article Title: Oral Sensory Neurons of the Geniculate Ganglion That Express Tyrosine Hydroxylase Comprise a Subpopulation That Contacts Type II and Type III Taste Bud Cells

doi: 10.1523/ENEURO.0523-21.2022

Figure Lengend Snippet: Th -expressing taste bud cells are a rare population of TRPM5+ Type II receptor cells. A , Fungiform (top row) or circumvallate (second row) taste buds from Th -CreER; Rosa RFP mice were immunolabeled for K8 (green), TUJ1 (blue), and RFP (magenta). Tissues were collected 3–4 d after the last tamoxifen injection. B , Fungiform taste buds from tamoxifen-treated Th -CreER; Rosa RFP mice were immunofluorescently labeled with antibodies to CAR4 (green) and RFP (magenta) or with antibodies to K8 (blue), RFP (magenta), and TRPM5 (blue). RFP+ cells did not overlap with CAR4 labeling (top row in B ) but did overlap with TRPM5 labeling (bottom row). C , Quantifications of the number of fungiform and circumvallate taste buds containing a RFP+ taste receptor cell. For fungiform taste buds, 42–49 taste buds were analyzed in each of three mice; for circumvallate taste buds 96–118 taste buds were evaluated each from three mice. Data are graphed as the mean ± SEM, n = 3, and p = 0.0012. Scale bars are 20 μm and ** indicates p < 0.01.

Article Snippet: The slides were then incubated with the following primary antibodies, which were diluted in PBS containing 0.3% Triton X-100 with 1% BSA in a humidified chamber overnight, or for 5 d (free floating tissues) at 4°C: rat α–cytokeratin-8 (TROMA-I, supernatant; 1:100 dilution, DSHB], rabbit α-RFP (1:200 dilution; Rockland, catalog #600-401-379), chicken α-RFP (1:200 dilution; Rockland, catalog #600-401-901), goat α-PHOX2B (1:200 dilution; R&D Systems, catalog #AF4940,), mouse α-TUJ1 (βIII-tubulin; 1:200 dilution; Sigma, catalog #T8578), goat α-CAR4 (1:200 dilution; R&D Systems, catalog #AF2414), and rabbit α-TRPM5 (1:200 dilution; Emily Liman’s Lab, University of Southern California).

Techniques: Expressing, Immunolabeling, Injection, Labeling

Journal: eNeuro

Article Title: Oral Sensory Neurons of the Geniculate Ganglion That Express Tyrosine Hydroxylase Comprise a Subpopulation That Contacts Type II and Type III Taste Bud Cells

doi: 10.1523/ENEURO.0523-21.2022

Figure Lengend Snippet: Fibers from Th -expressing neurons preferentially innervate taste buds in the soft palate and circumvallate papillae. A , Fungiform (top two rows), circumvallate (third row), anterior foliate (fourth row), soft palate (fifth row), and nasoincisor duct (sixth row) taste buds were immunolabeled with antibodies to K8 (green), TUJ1 (blue), and RFP (magenta). RFP+ fibers innervated half of fungiform, anterior foliate and nasoincisor duct taste buds, and innervated all circumvallate and soft palate taste buds. For fungiform taste buds, images are displayed in two separate rows depicting an example of innervated (top) and noninnervated (second row) taste buds. B , Chorda tympani and greater superficial petrosal (GSP) nerves were immunofluorescently labeled with antibodies to TUJ1 (green) and RFP (magenta), showing the presence of RFP+ fibers projecting through each nerve. Three mice were imaged in these experiments. Scale bars in A are 20 μm, and the scale bar in the top row of fungiform taste bud images applies to the second row as well. The scale bar in B is 100 μm and applies to all panels in B .

Article Snippet: The slides were then incubated with the following primary antibodies, which were diluted in PBS containing 0.3% Triton X-100 with 1% BSA in a humidified chamber overnight, or for 5 d (free floating tissues) at 4°C: rat α–cytokeratin-8 (TROMA-I, supernatant; 1:100 dilution, DSHB], rabbit α-RFP (1:200 dilution; Rockland, catalog #600-401-379), chicken α-RFP (1:200 dilution; Rockland, catalog #600-401-901), goat α-PHOX2B (1:200 dilution; R&D Systems, catalog #AF4940,), mouse α-TUJ1 (βIII-tubulin; 1:200 dilution; Sigma, catalog #T8578), goat α-CAR4 (1:200 dilution; R&D Systems, catalog #AF2414), and rabbit α-TRPM5 (1:200 dilution; Emily Liman’s Lab, University of Southern California).

Techniques: Expressing, Immunolabeling, Labeling

Journal: eNeuro

Article Title: Oral Sensory Neurons of the Geniculate Ganglion That Express Tyrosine Hydroxylase Comprise a Subpopulation That Contacts Type II and Type III Taste Bud Cells

doi: 10.1523/ENEURO.0523-21.2022

Figure Lengend Snippet: Fibers from Th + oral sensory neurons enter the rostral portion of the NTS. A , Th -CreER; Rosa RFP mice were administered tamoxifen and allowed to recover for three weeks before the brainstem was isolated. Coronal serial sections through the NTS were immunolabeled with antibodies to TUJ1 (green) and RFP (magenta). The white arrows indicate the solitary tract that is lateral to the NTS (indicated with yellow arrows). Successive sections are displayed progressing from top row to bottom row so that the progression of the solitary tract to the NTS can be appreciated. Scale bar: 100 μm. B , Th -CreER; Phox2b -FLPO; RC::FLTG mice were administered tamoxifen and allowed to recover for three weeks. Coronal serial sections were immunolabeled with antibodies to TUJ1 (blue) and eGFP (green). The white arrows indicate the solitary tract and yellow arrows denote the NTS. The bottom row is a higher power image of the row above it. The scale bars are 100 μm. The scale bar in the first row in A applies to all panels in A , and the scale bar in B applies to all panels in B . Four independent mice of each genotype were analyzed in A and B .

Article Snippet: The slides were then incubated with the following primary antibodies, which were diluted in PBS containing 0.3% Triton X-100 with 1% BSA in a humidified chamber overnight, or for 5 d (free floating tissues) at 4°C: rat α–cytokeratin-8 (TROMA-I, supernatant; 1:100 dilution, DSHB], rabbit α-RFP (1:200 dilution; Rockland, catalog #600-401-379), chicken α-RFP (1:200 dilution; Rockland, catalog #600-401-901), goat α-PHOX2B (1:200 dilution; R&D Systems, catalog #AF4940,), mouse α-TUJ1 (βIII-tubulin; 1:200 dilution; Sigma, catalog #T8578), goat α-CAR4 (1:200 dilution; R&D Systems, catalog #AF2414), and rabbit α-TRPM5 (1:200 dilution; Emily Liman’s Lab, University of Southern California).

Techniques: Isolation, Immunolabeling

Journal: bioRxiv

Article Title: Ectopic histone clipping in the mouse model of progressive myoclonus epilepsy

doi: 10.1101/2020.08.21.261180

Figure Lengend Snippet: H3T22 cleavage is enriched in TUJ1-expressing neurons. Representative immunofluorescence images of differentiating wt and Cstb −/− NPCs stained for TUJ1, H3T22cl and DNA (Hoechst). Scale bar = 20 μm. Bar plot depicts the proportion of TUJ1/H3T22cl -positive cells over total H3T22cl-positive cell count (n = 6).

Article Snippet: Primary antibodies used: rabbit α-H3 C-terminus (Abcam, Western blot (WB) 1:15000, RRID: AB_302613), rabbit α-H3T22cl (Cell Signaling Technologies, WB 1:1000, immunocytochemistry (ICC) 1:500, RRID: AB_2797961), rabbit α-H3K4me3 (Abcam, WB 1:2000, RRID: AB_306649), rabbit α-H3K9me3 (Abcam, WB 1:3000, RRID: AB_2797591), rabbit α-H3K27me2 (Abcam, WB 1:4000, RRID: AB_448222), mouse α-Nestin (Millipore, ICC 1:100, RRID: AB_94911), rabbit α-GFAP (Agilent Technologies, ICC 1:200, RRID: AB_10013382), rat α-GFAP (Thermo Fisher Scientific, ICC 1:500, RRID: AB_2532994), mouse α-DCX (Santa Cruz Biotechnology, ICC 1:150, RRID: AB_10610966), mouse α-βIII-tubulin (TuJ1) (Millipore, ICC 1:500, RRID: AB_2210524), mouse α-O4 (Millipore, ICC 1:200, RRID: AB_11213138), rat α-Ki-67 (Thermo Fisher Scientific, ICC 1:200, RRID: AB_10853185).

Techniques: Expressing, Immunofluorescence, Staining, Cell Counting

Journal: bioRxiv

Article Title: Ectopic histone clipping in the mouse model of progressive myoclonus epilepsy

doi: 10.1101/2020.08.21.261180

Figure Lengend Snippet: Changes in the relative production of neurons and astrocytes upon NPC differentiation in “astrocyte media” and “neuron media”. Representative immunofluorescence images of differentiating wt NPCs cultured for 12 days under neurogenesis-promoting conditions (neuron media) and astrogliogenesis-promoting conditions (astrocyte media) and stained for GFAP and TUJ1. Scale bar = 50 μm. Bar plot depicts the proportion of GFAP-positive and TUJ1-positive cells over total cell count at day 12 pD (n = 4-5). * p < 0.05, *** p < 0.001.

Article Snippet: Primary antibodies used: rabbit α-H3 C-terminus (Abcam, Western blot (WB) 1:15000, RRID: AB_302613), rabbit α-H3T22cl (Cell Signaling Technologies, WB 1:1000, immunocytochemistry (ICC) 1:500, RRID: AB_2797961), rabbit α-H3K4me3 (Abcam, WB 1:2000, RRID: AB_306649), rabbit α-H3K9me3 (Abcam, WB 1:3000, RRID: AB_2797591), rabbit α-H3K27me2 (Abcam, WB 1:4000, RRID: AB_448222), mouse α-Nestin (Millipore, ICC 1:100, RRID: AB_94911), rabbit α-GFAP (Agilent Technologies, ICC 1:200, RRID: AB_10013382), rat α-GFAP (Thermo Fisher Scientific, ICC 1:500, RRID: AB_2532994), mouse α-DCX (Santa Cruz Biotechnology, ICC 1:150, RRID: AB_10610966), mouse α-βIII-tubulin (TuJ1) (Millipore, ICC 1:500, RRID: AB_2210524), mouse α-O4 (Millipore, ICC 1:200, RRID: AB_11213138), rat α-Ki-67 (Thermo Fisher Scientific, ICC 1:200, RRID: AB_10853185).

Techniques: Immunofluorescence, Cell Culture, Staining, Cell Counting

Journal: Development (Cambridge, England)

Article Title: SOX2 is required for inner ear growth and cochlear nonsensory formation before sensory development

doi: 10.1242/dev.170522

Figure Lengend Snippet: Analysis of NEUROG1-deficient ears indicates that the nonsensory defects are not caused secondarily from reductions in the otic ganglion. (A-C) Low-power midmodiolar sections from control (A), Neurog1−/− (B) and E8.5 Sox2-deleted cochleae (C). The nonsensory phenotype was more severe in the absence of E8.5 SOX2 than of NEUROG1, and some neuronal formation still occurred [arrows mark neuronal expression (TUJ1; white) in C,F,I]. (D-K) Higher magnification views of the boxed regions in A-C highlight that sensory formation occurred fairly normally in the Neurog1-deficient mutant. (L-T) Examples of cross-sections through each vestibular organ in control (L-O), Neurog1−/− mutant (P-S) and E8.5 SOX2-deleted inner ears (T). The vestibule forms relatively normally in Neurog1-deficient mutants, with the exception of a smaller saccule devoid of sensory markers (arrow, P). By contrast, the only vestibular structure seen in E8.5 SOX2-deficient inner ears was an underdeveloped saccule (T). Scale bars: 50 µm.

Article Snippet: The primary antibodies used were goat polyclonal α-SOX2 (1:700, Santa Cruz Biotechnology, sc-17320), rat monoclonal α-RFP (1:1000, ChromoTek, 5F8), rabbit polyclonal α-MYO6 (1:700, Proteus BioSciences, 25-6791), mouse monoclonal α-TUJ1 (1:1000, Covance, MMS-435P), rabbit polyclonal α-pHH3 [1:400, Santa Cruz Biotechnology, Ser10(6G3)], mouse monoclonal α-pHH3 (1:400, Cell Signaling Technology, sc-8656-R), rabbit polyclonal α-cleaved caspase3 (1:1000, R&D Systems: AF835), goat polyclonal α-JAG1 c-20 (1:200, Santa Cruz Biotechnology, SC-6011) and mouse polyclonal α-CDKN1B/p27 Kip1 (1:100, Thermo Fisher Scientific, MS-256-P).

Techniques: Expressing, Mutagenesis