βii tubulin  (R&D Systems)

Journal: medRxiv

Article Title: FRMPD4 , a causal gene for intellectual disability and epilepsy, is associated with X-linked non-syndromic hearing loss

doi: 10.64898/2026.03.27.26349271

Figure Lengend Snippet: Loss of function of frmpd4 in zebrafish results in neuromast alterations in the otic vesicle and in the posterior lateral line. (A) Functional test of frmpd4 splice blocking Morpholino via RT-PCR (per condition 10 pooled embryos). Exon-spanning PCR primers indicate the loss of a correctly spliced frmpd4 band after Morpholino injection, and gain of a larger, unspliced additional PCR product. PCR amplification of ef1a1 were used as internal cDNA control. (B) DASPEI staining for neuromasts in frmpd4 Morpholino injected knockdown embryos. Quantification of neuromast number in the posterior lateral line (C) , the otic vesicle (D) and the preoptical/supraorbital (E) indicated reduction after frmpd4 knockdown (0.25mM Morpholino concentration, age: 4 dpf, comparison to embryos injected with standard MO). (F) Measurement of neuromast distance in the posterior lateral line indicated no significant change after frmpd4 Morpholino knockdown. (G) Staining for acetyl tubulin after frmpd4 Morpholino knockdown in 4 dpf larvae indicated similar to in frmpd4 sa12377 mutants loss of neuronal cells and axonal projections in ventral sensory patches of the otic vesicle and interference with correct neuromast formation in the posterior lateral line. Analyzed embryos: 2 wild type controls; 4 frmpd4 MO 0.25mM; 2 frmpd4 MO 0.5mM.

Article Snippet: Afterwards, the tissue was incubated with a primary antibody solution containing 3% normal horse serum, 1% bovine serum albumin, 0.3% Triton X-100, and 0.1% Tween20 with primary antibodies for βII-tubulin (mouse monoclonal, R&D System, #MAB1192 clone Tuj1, dilution 1:1000) and FRMPD4 (rabbit, polyclonal, dilution 1:500) ( ) incubated overnight at 4°C.

Techniques: Functional Assay, Blocking Assay, Reverse Transcription Polymerase Chain Reaction, Injection, Amplification, Control, Staining, Knockdown, Concentration Assay, Comparison

Journal: medRxiv

Article Title: FRMPD4 , a causal gene for intellectual disability and epilepsy, is associated with X-linked non-syndromic hearing loss

doi: 10.64898/2026.03.27.26349271

Figure Lengend Snippet: frmpd4 CRISPant in zebrafish show only mild neuromast alterations in the otic vesicle and in the posterior lateral line. (A) DASPEI staining for neuromasts in CRISPants embryos (F0 generation, frmpd4 sgRNA r2 and f3 injected). Quantification of neuromast number in the posterior lateral line (B) , the otic vesicle (C) , and the preoptical/supraorbital (D) indicated reduction in frmpd4 CRISPants (4 dpf). (E) Measurement of neuromast distance in the posterior lateral line indicated no significant change after frmpd4 CRISPR or Morpholino knockdown. (F) Staining for acetylated tubulin in 4 dpf embryos indicated mild changes to neuronal cells and axonal projections in ventral sensory patches of the otic vesicle and not significant interference with correct neuromast spacing in the posterior lateral line after transient frmpd4 CRISPR knockdown. Analyzed embryos: 2 wild type controls; 4 frmpd4 CRISPants.

Article Snippet: Afterwards, the tissue was incubated with a primary antibody solution containing 3% normal horse serum, 1% bovine serum albumin, 0.3% Triton X-100, and 0.1% Tween20 with primary antibodies for βII-tubulin (mouse monoclonal, R&D System, #MAB1192 clone Tuj1, dilution 1:1000) and FRMPD4 (rabbit, polyclonal, dilution 1:500) ( ) incubated overnight at 4°C.

Techniques: Staining, Injection, CRISPR, Knockdown

Journal: medRxiv

Article Title: FRMPD4 , a causal gene for intellectual disability and epilepsy, is associated with X-linked non-syndromic hearing loss

doi: 10.64898/2026.03.27.26349271

Figure Lengend Snippet: Loss of function of frmpd4 in zebrafish results in axonal and structural malformations in the otic vesicle and the posterior lateral line. (A) Staining for acetylated tubulin in 4 dpf embryos indicated loss of neuronal cell and axonal projection in ventral sensory patches of the otic vesicle in homozygous frmpd4 sa12377/sa12377 mutants. (B) Posterior lateral line neuromasts and axons (white arrowheads) are affected by frmpd4 loss by depicting size reduction and morphological changes (analyzed embryos per genotype: 7 wild type controls; 6 heterozygous frmpd4 sa12377/+ ; 5 homozygous frmpd4 sa12377/sa12377 ). (C) Neuromast cell deposition in the PLL can be disrupted in homozygous frmpd4 sa12377/sa12377 mutants, while adjacent somites show normal patterns (n=3 embryos per genotype; control and heterozygous frmpd4 sa12377/+ mutants display indistinguishable patterns). (D) Scanning electron microscopy further showed disruption of cellular organization in PLL neuromasts and loss of kinocilia in in frmpd4 sa12377/sa12377 mutants (n=3 per genotype). m: anterior macula, pm: posterior macula, ac: anterior crista, lc: lateral crista, pc: posterior crista. Scale bars in A to C indicate 50 µm.

Article Snippet: Afterwards, the tissue was incubated with a primary antibody solution containing 3% normal horse serum, 1% bovine serum albumin, 0.3% Triton X-100, and 0.1% Tween20 with primary antibodies for βII-tubulin (mouse monoclonal, R&D System, #MAB1192 clone Tuj1, dilution 1:1000) and FRMPD4 (rabbit, polyclonal, dilution 1:500) ( ) incubated overnight at 4°C.

Techniques: Staining, Control, Electron Microscopy, Disruption

Journal: Journal of Alzheimer's Disease Reports

Article Title: Tubulin Isotypes and Posttranslational Modifications in Vascular Dementia and Alzheimer’s Disease

doi: 10.3233/ADR-220068

Figure Lengend Snippet: Levels of various tubulin isotypes in the frontal and temporal lobes of human brain homogenates. A) Human brain homogenates were probed against α-tubulin, βII-tubulin, and βIII- tubulin. Total protein was used for normalization. B) Immunoblot quantifications in the frontal lobe and (C) immunoblot quantifications in the temporal lobe. 10μg of protein was added on each lane. Data are expressed as the relative ratio of protein/total protein and graphs represent individual values with mean±SEM. One-Way ANOVA with Tukey’s posthoc tests. * p < 0.05, ** p < 0.01. N for frontal lobe: C (4–7), V (7-8), A (5–7). N for the temporal lobe: C (13), V (14), A (15). C, controls; V/VaD, vascular dementia; A/AD, Alzheimer’s disease. Please note that the number of analyzed samples (N) for the frontal lobe varies depending on samples available for the assay.

Article Snippet: The other antibodies used in the current project were purchased from different companies: βII-tubulin (1:2000; Genetex, GTX100117), βIII-tubulin (1:5000; Biolegend, 801202), acetyl-α-tubulin Lys40 (1:2000; Elabsciences, E-AB-20302), MAP2 (1:2000; Proteintech, 17490-1-AP), MAP6 (1:500; Scientific LTD, ARP60316-P050), PolyE (1:4000; AdipoGen Life Sciences, AG-25B-0030-CO50), and tau (1:10000; Proteintech, 66499-1-Ig).

Techniques: Western Blot

Journal: Journal of Alzheimer's Disease Reports

Article Title: Tubulin Isotypes and Posttranslational Modifications in Vascular Dementia and Alzheimer’s Disease

doi: 10.3233/ADR-220068

Figure Lengend Snippet: Levels of various tubulin PTMs in the frontal and temporal lobes of human brain homogenates. A) Human brain homogenates were probed against PolyE, Tyr, DeTyr, and Ac.tub. Total protein was used for normalization. B) Immunoblot quantifications in the frontal lobe and (C) immunoblot quantifications in the temporal lobe. 10μg of protein was added on each lane. Data are expressed as the relative ratio of protein/total protein and graphs represent individual values with mean±SEM. One-Way ANOVA with Tukey’s posthoc tests. * p < 0.05, ** p < 0.01. N for frontal lobe: C (5–7), V (7-8), A (6-7). N for the temporal lobe: C (13), V (14), A (15). C, controls; V/VaD, vascular dementia; A/AD, Alzheimer’s disease. Please note that the number of analyzed samples (N) for the frontal lobe varies depending on samples available for the assay.

Article Snippet: The other antibodies used in the current project were purchased from different companies: βII-tubulin (1:2000; Genetex, GTX100117), βIII-tubulin (1:5000; Biolegend, 801202), acetyl-α-tubulin Lys40 (1:2000; Elabsciences, E-AB-20302), MAP2 (1:2000; Proteintech, 17490-1-AP), MAP6 (1:500; Scientific LTD, ARP60316-P050), PolyE (1:4000; AdipoGen Life Sciences, AG-25B-0030-CO50), and tau (1:10000; Proteintech, 66499-1-Ig).

Techniques: Western Blot

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Soft Matrix Combined With BMPR Inhibition Regulates Neurogenic Differentiation of Human Umbilical Cord Mesenchymal Stem Cells

doi: 10.3389/fbioe.2020.00791

Figure Lengend Snippet: Matrix stiffness regulates cell morphology and neural differentiation ofhuman umbilicalcordmesenchymal stem cells (hUC-MSCs). (A) A summary of substrate stiffness construction. Matrixes of different stiffness were prepared by mixing the same concentration of acrylamide (AAm) with different concentrations of bis-acrylamide (Bis-AAm; 0.05, 0.3, and 0.7%). Sulfo-SANPAH (hexanoate) was cross-linked via ultraviolet irradiation and then coated with fibronectin for cell adhesion. (B) Matrix stiffness affects the morphological characteristics of hUC-MSCs. Morphological characteristics of hUC-MSCs were observed under a scanning electron microscope at days 1 and 7. TCP group: cells on Tissue Culture Plate. Scale bar = 20 μm. n = 3 independent wells. (C) The cell aspect ratio was calculated using NIH Image J. The aspect ratio of the cell is the ratio of the major to minor axes ( n = 3, * p < 0.05,and ** p < 0.01). (D) The cell area was calculated using NIH Image J ( n = 3, * p < 0.05,and ** p < 0.01). (E) qRT-PCR analyses were performed to detect the expression of neuronal-specific markers Nestin and βIII-tubulin in hUC-MSCs at day 1 and day 7 (Results are presented as mean ± SEM, n = 5 independent experiments, * p < 0.05, and ** p < 0.01). (F) Western blotting analysis of Nestin and βIII-tubulin at day 7 (Results are presented as mean ± SEM, n = 3, * p < 0.05, and ** p < 0.01). (G) qRT-PCR analysis of stem cells self-renewal markers SOX2 and OCT4 at day 1 (Results are presented as mean ± SEM, n = 5 independent experiments, * p < 0.05, ** p < 0.01, and *** p < 0.001).

Article Snippet: Afterwards, the membranes were blocked and probed with antibodies for BMPRIA (Abcam), BMPRIB (Abcam), BMPRII (Abcam), GAPDH (CST), Nestin (CST), βII-tubulin (CST), GSK-3β (CST), and FAK (CST) overnight at 4°C.

Techniques: Concentration Assay, Irradiation, Microscopy, Quantitative RT-PCR, Expressing, Western Blot

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Soft Matrix Combined With BMPR Inhibition Regulates Neurogenic Differentiation of Human Umbilical Cord Mesenchymal Stem Cells

doi: 10.3389/fbioe.2020.00791

Figure Lengend Snippet: BMPR is involved in stiffness-mediated neural differentiation of hUC-MSCs. (A) qRT-PCR detection of BMPR subtypes expression at day 1 and day 7 (Results are presented as mean ± SEM, n = 5 independent experiments, * p < 0.05, and ** p < 0.01). (B) Expression of BMPR subtypes detected using western blotting, and statistical analysis diagram (Results are presented as mean ± SEM, n = 3, * p < 0.05, and ** p < 0.01). (C) The best concentration of BMPR inhibitor was determined using western blotting, the best concentration of inhibitor was 2 μM ( n = 3 independent experiments). (D) The cells were collected at day 7 for detection of BMPR expression after stiffness groups adding inhibitor using RT-qPCR (Results are presented as mean ± SEM, n = 5 independent experiments, * p < 0.05, and ** p < 0.01). (E) Expression of p-SMAD was detected using western blotting before and after BMPR inhibition, and statistical analysis diagram (Results are presented as mean ± SEM, n = 3, * p < 0.05, and ** p < 0.01). (F) qRT-PCR analysis of Nestin and βIII-tubulin expression after BMPR inhibition for 24 h. Statistical analysis was performed using one-way ANOVA. Statistical significance was defined as p < 0.05. Results are presented as mean ± SEM ( n = 5 independent experiments, p < 0.05). (G) qRT-PCR analysis of SOX2 and OCT4 expression after BMPR inhibition for 24 h. Statistical analysis was performed using one-way ANOVA. Statistical significance was defined as p < 0.05. Results are presented as mean ± SEM ( n = 5 independent experiments, p < 0.05, and *** p < 0.001).

Article Snippet: Afterwards, the membranes were blocked and probed with antibodies for BMPRIA (Abcam), BMPRIB (Abcam), BMPRII (Abcam), GAPDH (CST), Nestin (CST), βII-tubulin (CST), GSK-3β (CST), and FAK (CST) overnight at 4°C.

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Concentration Assay, Inhibition

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Soft Matrix Combined With BMPR Inhibition Regulates Neurogenic Differentiation of Human Umbilical Cord Mesenchymal Stem Cells

doi: 10.3389/fbioe.2020.00791

Figure Lengend Snippet: Primer sequences for qRT-PCR.

Article Snippet: Afterwards, the membranes were blocked and probed with antibodies for BMPRIA (Abcam), BMPRIB (Abcam), BMPRII (Abcam), GAPDH (CST), Nestin (CST), βII-tubulin (CST), GSK-3β (CST), and FAK (CST) overnight at 4°C.

Techniques:

Journal: American Journal of Physiology - Cell Physiology

Article Title: Altered skeletal muscle microtubule-mitochondrial VDAC2 binding is related to bioenergetic impairments after paclitaxel but not vinblastine chemotherapies

doi: 10.1152/ajpcell.00384.2018

Figure Lengend Snippet: α-Tubulin- voltage-dependent anion channel (VDAC)2 interactions in extensor digitorum longus (EDL) single fibers increase following paclitaxel stabilization. A: model of tubulin regulation of VDAC-dependent ADP/ATP exchange (9). Confocal microscopy images of single EDL fiber. B: control experiments in the presence of either α-tubulin, βII-tubulin, or VDAC2, incubated alone or together (n = 3) C: proximity ligation assay representative images. D: graphical depiction of α-tubulin-VDAC2 protein interaction (n = 8, *P = 0.001 vs. control). E: graphical depiction of βII-tubulin-VDAC2 interaction (n = 8–9, *P = 0.0001 vs. control). Scale bar, 16 μm. Results are reported as means ± SEM.

Article Snippet: Primary and secondary antibodies used were as follows: α-tubulin (1:1,000, Sigma-Aldrich; T6199), βII-tubulin (1:250, Abcam; ab28036; separate fibers), VDAC2 (1:250, Santa Cruz; 32059), Alexa Fluor 488 (Invitrogen/Thermo-Fisher Scientific; A21121), and Alexa Fluor 555 (Invitrogen/Thermo-Fisher Scientific; AS1431).

Techniques: Confocal Microscopy, Incubation, Proximity Ligation Assay

Journal: American Journal of Physiology - Cell Physiology

Article Title: Altered skeletal muscle microtubule-mitochondrial VDAC2 binding is related to bioenergetic impairments after paclitaxel but not vinblastine chemotherapies

doi: 10.1152/ajpcell.00384.2018

Figure Lengend Snippet: α-Tubulin but not βII-tubulin organization is altered following microtubule-targeted chemotherapy. Confocal microscopy representative images of single extensor digitorum longus fibers stained with antibodies recognizing α-tubulin and βII-tubulin following either control (DMSO), paclitaxel, or vinblastine treatment. Scale bar, 16 μm.

Article Snippet: Primary and secondary antibodies used were as follows: α-tubulin (1:1,000, Sigma-Aldrich; T6199), βII-tubulin (1:250, Abcam; ab28036; separate fibers), VDAC2 (1:250, Santa Cruz; 32059), Alexa Fluor 488 (Invitrogen/Thermo-Fisher Scientific; A21121), and Alexa Fluor 555 (Invitrogen/Thermo-Fisher Scientific; AS1431).

Techniques: Confocal Microscopy, Staining