Journal: BMC Cancer

Article Title: Tubulin binding cofactor C (TBCC) suppresses tumor growth and enhances chemosensitivity in human breast cancer cells

doi: 10.1186/1471-2407-10-135

Figure Lengend Snippet: Effect of tubulin binding cofactor C (TBCC) on the expression level of related proteins . Western blot analysis of total cell extracts of the clones showing the effect of overexpression of TBCC on the expression level of TBCC, TBCD, Arl2 and p53. β-actin was used as a reference protein. MC+1, MC+2 and MC+3 represent MCF7 cells stably transfected with pcDNA6/C+. MP6.1, MP6.2 and MP6.3 represent MCF7 cells stably transfected with empty pcDNA6.

Article Snippet: The antibodies used were anti β III-tubulin (clone Tuj1, 1/2500; Covalab, Lyon, France), anti p53 (clone DO7, 1/1000; Dako, Denmark), anti β-actin (clone AC-15, 1/5000), anti α-tubulin (clone DM1A, 1/1000), anti β-tubulin (clone 2.1, 1/1000), anti tyrosinated α-tubulin (clone TUB-1A2, 1/1000) and acetylated α-tubulin (clone 6-11B-1, 1/1000) from Sigma Aldrich (St Quentin Fallavier, France).

Techniques: Binding Assay, Expressing, Western Blot, Clone Assay, Over Expression, Stable Transfection, Transfection

Journal: BMC Cancer

Article Title: Tubulin binding cofactor C (TBCC) suppresses tumor growth and enhances chemosensitivity in human breast cancer cells

doi: 10.1186/1471-2407-10-135

Figure Lengend Snippet: Localization of TBCC in MC+1 and MP6.1 cells . (A) Representative images of MC+1 and MP6.1 cells after DNA (DAPI, blue) and β-tubulin (FITC, green) staining (B) Representative images of MC+1 and MP6.1 cells after DNA (DAPI, blue) and TBCC (FITC, green) staining (C) Representative images of DNA (DAPI, blue) and TBCC (FITC, green) staining in MC+1 cells exposed 24 hours to 10 nM of paclitaxel.

Article Snippet: The antibodies used were anti β III-tubulin (clone Tuj1, 1/2500; Covalab, Lyon, France), anti p53 (clone DO7, 1/1000; Dako, Denmark), anti β-actin (clone AC-15, 1/5000), anti α-tubulin (clone DM1A, 1/1000), anti β-tubulin (clone 2.1, 1/1000), anti tyrosinated α-tubulin (clone TUB-1A2, 1/1000) and acetylated α-tubulin (clone 6-11B-1, 1/1000) from Sigma Aldrich (St Quentin Fallavier, France).

Techniques: Staining

Journal: BMC Cancer

Article Title: Tubulin binding cofactor C (TBCC) suppresses tumor growth and enhances chemosensitivity in human breast cancer cells

doi: 10.1186/1471-2407-10-135

Figure Lengend Snippet: Effect of TBCC on different tubulins content and on subcellular tubulin fractions . (A) Western blot analysis of total cell extracts of the MC+1, MC+2, MC+3, MP6.1, MP6.2 and MP6.3 clones showing the effect of overexpression of TBCC on the expression level of α-tubulin, β-tubulin, tyrosinated α-tubulin, detyrosinated (glu) α-tubulin, β III-tubulin and acetylated α-tubulin. β-actin was used as a reference protein. (B) Representative blots corresponding to the expression levels of α-tubulin and β-tubulin in nonpolymerizable tubulin heterodimers (NPT), polymerizable tubulin (PT) heterodimers, microtubule heterodimers (MT) and total pool of tubulins in MC+1 and MP6.1 cells. These fractions were obtained after series of ultracentrifugations. (C) Protein ratios (MC+1 vs. MP6.1) of both α-tubulins and β-tubulins levels in NPT, PT and MT fractions. Results presented are the average values of three experiments. Bars represent standard deviation.

Article Snippet: The antibodies used were anti β III-tubulin (clone Tuj1, 1/2500; Covalab, Lyon, France), anti p53 (clone DO7, 1/1000; Dako, Denmark), anti β-actin (clone AC-15, 1/5000), anti α-tubulin (clone DM1A, 1/1000), anti β-tubulin (clone 2.1, 1/1000), anti tyrosinated α-tubulin (clone TUB-1A2, 1/1000) and acetylated α-tubulin (clone 6-11B-1, 1/1000) from Sigma Aldrich (St Quentin Fallavier, France).

Techniques: Western Blot, Clone Assay, Over Expression, Expressing, Standard Deviation

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

Journal: STAR Protocols

Article Title: Protocol for culturing neurospheres from progenitor cells in the dentate gyrus of aged mouse hippocampus

doi: 10.1016/j.xpro.2025.103692

Figure Lengend Snippet:

Article Snippet: Polyclonal rabbit IgG anti-β-Tubulin III/Tuj1 (dilution 1:500) , GenScript , Cat#A01627; RRID: AB_2622164.

Techniques: Recombinant, Microscopy, Electron Microscopy, Sterility, Cell Culture, Software

Journal: bioRxiv

Article Title: Oncogenic mutations disrupt lineage differentiation via epigenetic perturbations

doi: 10.1101/2025.04.22.649974

Figure Lengend Snippet: a, Experimental scheme for generating induced neuronal (iN) cells differentiated from human embryonic stem cells (hESCs) or neural progenitor cells (NPCs) by overexpression of the neuronal gene, Ngn2. b, Brightfield and fluorescent images of iN cells stained for the neural cell marker Tuj1 (red), pluripotency marker Sox2 (red), cell proliferation marker Ki67 (green) from wt hESC-derived iN cells and mutant hESC-derived iN cells after 2 weeks of differentiation. Data are from at least three independent experiments. Scale bars, 50 μm. c, Neurite length in cells were measured at 12 days of iN differentiation (*p < 0.01). d, Quantitative RT-PCR analysis of neural cell marker Map2 expression level at 7 days of hESC differentiation. Two-tailed t-test, Mean ± SD (n=3, *p < 0.01). e, Brightfield and fluorescent images of NPC-derived iN cells stained for the neural cells marker Tuj1 (green) from IDH1-wt NPC-derived iN cells and IDH1-R132H mutation NPC-derived iN cells after 2 weeks of differentiation. f, Quantitative RT-PCR analysis of neural cell marker Map2 expression at 7 days of NPC differentiation. Values represent mean ± SD (n=3, *p < 0.01). g, Cell number counting of Ki67 positive wt hESC-derived iN cells and mutant hESC-derived iN cells at 10 days of differentiation (*p < 0.01). h, Cell number counting of Tuj1 positive wt hESC-derived iN cells and mutant hESC-derived iN cells at 10 days of differentiation (*p < 0.01). i, Cell number counting of Sox2 positive wt hESC-derived iN cells and mutant hESC-derived iN cells at 10 days of differentiation. Values represent the number of cells showing positive in each situation ± SD (n=3, *p < 0.01).

Article Snippet: The following primary antibodies with indicated dilution in blocking buffer were used: Rabbit anti-Tuj1 (Covance, BioLegend, MRB-435P, 1:1,000), Mouse anti-Tuj1 (Covance, BioLegend, MMS-435P, 1:1,000), Rabbit anti-human Sox1 (Millipore, AB15766, 1:1000), Mouse anti-Tra-1-60 (Millipore, MAB4360, 1:1000), Rabbit anti-Ki67 (Abcam, ab15580, 1:1000), Goat anti-Sox2 (Santa Cruz, Y-17, sc-17320, 1:1000), Mouse anti-Sox2 (R&D, MAB2018, 1:200), Goat anti-Sox2 (R&D, AF2018, 1:200), Mouse anti-Oct-3/4 (Sata Cruz, C-10, sc-5279), Mouse anit-Nestin (DSHB, rat-401, 1:200), Mouse anit-Nestin (R&D, MAB1259, 1:200), Rabbit anti-ASCL1 (Abcam, ab74065, 1:200).

Techniques: Over Expression, Staining, Marker, Derivative Assay, Mutagenesis, Quantitative RT-PCR, Expressing, Two Tailed Test

Journal: bioRxiv

Article Title: Oncogenic mutations disrupt lineage differentiation via epigenetic perturbations

doi: 10.1101/2025.04.22.649974

Figure Lengend Snippet: a, Cell number counting of wt hESCs and mutant hESCs after 3 days of dox-induction (n=3, NS: p > 0.05). b, Cell number counting of wt hESC-derived iN cells and mutant hESC-derived iN cells at 7 days of differentiation (n=3, *p < 0.01). c, The ratio of the Sox2 positive hESC-derived iN cell number and Tuj1 positive hESC-derived iN cell number at 10 days of differentiation (*p < 0.01).

Article Snippet: The following primary antibodies with indicated dilution in blocking buffer were used: Rabbit anti-Tuj1 (Covance, BioLegend, MRB-435P, 1:1,000), Mouse anti-Tuj1 (Covance, BioLegend, MMS-435P, 1:1,000), Rabbit anti-human Sox1 (Millipore, AB15766, 1:1000), Mouse anti-Tra-1-60 (Millipore, MAB4360, 1:1000), Rabbit anti-Ki67 (Abcam, ab15580, 1:1000), Goat anti-Sox2 (Santa Cruz, Y-17, sc-17320, 1:1000), Mouse anti-Sox2 (R&D, MAB2018, 1:200), Goat anti-Sox2 (R&D, AF2018, 1:200), Mouse anti-Oct-3/4 (Sata Cruz, C-10, sc-5279), Mouse anit-Nestin (DSHB, rat-401, 1:200), Mouse anit-Nestin (R&D, MAB1259, 1:200), Rabbit anti-ASCL1 (Abcam, ab74065, 1:200).

Techniques: Mutagenesis, Derivative Assay

Journal: bioRxiv

Article Title: Oncogenic mutations disrupt lineage differentiation via epigenetic perturbations

doi: 10.1101/2025.04.22.649974

Figure Lengend Snippet: a, The morphology of U251 cells. U251 cell line is an IDH1 mutation glioblastoma cell line. Scale bars, 25 μm. b, c, Brightfield and fluorescent images of immunostaining TRA-1-60 (red) of U251 cells at 14 days reprogramming with OSKM for colony identification. d, e, Brightfield and fluorescent images of immunostaining U251 cells at 14 days differentiation by Ngn2 overexpression for the neural cells marker Tuj1 (green). f, The morphology of K562 cells. K562 cell line is an IDH1 mutant leukemia cell line. g, h, Brightfield and fluorescent images of K562 cells immunostaining for TRA-1-60 (green) at 14 days reprogramming with OSKM. No TRA-1-60 positive staining colony. i, j, Brightfield and fluorescent images of staining Tuj1 (red) and DAPI (blue) on K562 cells at 14 days differentiation by Ngn2 overexpression. k, The morphology of HCT116 cells. HCT116 cell line is an IDH1 mutant colon cancer cell line. l, m, Brightfield and fluorescent images of staining TRA-1-60 (green) and DAPI (blue) on HCT116 cells at 14 days reprogramming with OSKM. HCT116 cells cluster on feeders and grow more as colony-like clumps as shown in the photo. All cells are TRA-1-60 positive. n, o, Brightfield and fluorescent images of staining Tuj1 (red) and DAPI (blue) on HCT116 cells at 14 days differentiation by Ngn2 overexpression. p, q, r, s, Immunostaining U251 cells with the pluripotency marker Sox2 (red), neural stem/progenitor cell markers Nestin (green) and ASCL1 (red), and neural cell marker Tuj1 (green).

Article Snippet: The following primary antibodies with indicated dilution in blocking buffer were used: Rabbit anti-Tuj1 (Covance, BioLegend, MRB-435P, 1:1,000), Mouse anti-Tuj1 (Covance, BioLegend, MMS-435P, 1:1,000), Rabbit anti-human Sox1 (Millipore, AB15766, 1:1000), Mouse anti-Tra-1-60 (Millipore, MAB4360, 1:1000), Rabbit anti-Ki67 (Abcam, ab15580, 1:1000), Goat anti-Sox2 (Santa Cruz, Y-17, sc-17320, 1:1000), Mouse anti-Sox2 (R&D, MAB2018, 1:200), Goat anti-Sox2 (R&D, AF2018, 1:200), Mouse anti-Oct-3/4 (Sata Cruz, C-10, sc-5279), Mouse anit-Nestin (DSHB, rat-401, 1:200), Mouse anit-Nestin (R&D, MAB1259, 1:200), Rabbit anti-ASCL1 (Abcam, ab74065, 1:200).

Techniques: Mutagenesis, Immunostaining, Over Expression, Marker, Staining