ac tub  (Cell Signaling Technology Inc)

Journal: Nature Communications

Article Title: Capturing disease severity in LIS1-lissencephaly reveals proteostasis dysregulation in patient-derived forebrain organoids

doi: 10.1038/s41467-025-64980-0

Figure Lengend Snippet: A Bar graphs of –log₁₀-transformed p -values from GO molecular function enrichment for actin binding, actin filament binding, tubulin binding, and cytoskeleton structural constituent; nominal significance p = 0.05; p -value adjusted with Bonferroni correction; two-sided Hypergeometric test. B Representative day 20 organoids stained for acetylated α-tubulin (Ac-TUB); yellow dashed lines, apical/basal VZ regions. C Quantification of Ac-TUB strand density at day 20: n = 11 control (4 cell lines), 10 mild, 12 moderate, 18 severe (2 cell lines each), across four differentiations. VZ loops: 41, 54, 49, 80, respectively; two-sided Wilcoxon test. D Day 20 organoids stained for N-cadherin (N-CAD); yellow dashed lines, VZ edges and N-cadherin diameter expansion. E Quantification of apical N-CAD signal at day 20: VZ structures n = 32 control, 39 mild, 42 moderate, 43 severe for a total of 6, 8, 7, and 9 differentiations, respectively; 2 cell lines each; two-sided Kruskal–Wallis with post hoc Wilcoxon correction. F GO enrichment for cadherin binding; nominal significance p = 0.05; p -value adjusted with Bonferroni correction; two-sided Hypergeometric test. G Heatmap of log₂FC for WNT pathway genes in cycling progenitors by severity grade versus control (* P < 0.05, ** P < 0.01, *** P < 0.001); two-sided Wilcoxon test. H WNT-GFP reporter organoids at day 20; yellow dashed lines, VZ boundaries. I Mean value of WNT-GFP intensity per VZ: n = 20 control (2 cell lines), 20 mild (2 cell lines), 10 moderate (one cell line), 10 severe (one cell line); 3 organoids per condition from two differentiations per cell line; two-sided Kruskal–Wallis with post hoc Wilcoxon correction. J Quantification of cell division plane orientation: n = 13 control, 12 mild, 12 moderate, 15 severe. VZ loops: 59, 56, 43, 57; cells: 206, 359, 172, 249, respectively; Chi-square test. K Examples of division planes in control and severe organoids; yellow dashed lines, VZ edges; white dashed lines, division plane orientation. Scale bars: B left 200 μm, B right, H 50 μm; D , K 20 μm; All boxplots show median, 25th/75th percentiles, and 1.5 interquartile range whiskers. Source data are provided as a Source Data file.

Article Snippet: Primary antibodies were diluted according to the manufacturer’s instructions and incubated overnight at 4 °C with the following dilutions: Ac-TUB (1:500, Cell Signaling, cat. no. 5335S), AFP (1:200, Sino Biologicals, cat.no.

Techniques: Transformation Assay, Binding Assay, Staining, Control

Journal: Neural Regeneration Research

Article Title: ATAT1 deficiency enhances microglia/macrophage-mediated erythrophagocytosis and hematoma absorption following intracerebral hemorrhage

doi: 10.4103/1673-5374.382984

Figure Lengend Snippet: The expression of Ac-α-tub is significantly reduced in phagocytes. (A) Schematic of the co-culture of RAW264.7/BV2 cells with RBCs in vitro and experimental analyses. (B) Representative images of GFP-positive RAW264.7 cells co-cultured with DiO-labeled RBCs (scale bar: 10 µm). White arrow indicates the phagocytosed RBC. (C, D) Representative western blot (C) and quantification (D) of Ac-α-tub and α-tubulin in the FACS-sorted GFP-negative and GFP-positive RAW264.7/BV2 cells. (E, F) Representative immunofluorescence images (E) and quantification (F) of Ac-α-tub in control RAW264.7 cells and RAW264.7 cells co-cultured with unlabeled RBCs (scale bar: 10 µm). Data are shown as mean ± SEM ( n = 5 independent cell cultures). ** P < 0.01, *** P < 0.001, vs . GFP negative or control (two-tailed Student’s t -test). Ac-α-tub: Acetylated α-tubulin; GFP: green fluorescent protein; MFI: mean fluorescence intensity; RBCs: red blood cells.

Article Snippet: After blocking in 3% BSA for 2 hours, the membranes were incubated with the following primary antibodies overnight at 4°C: mouse anti-α-tubulin (1:1000, Abcam, Cambridge, UK, Cat# ab7291, RRID: AB_2241126) and rabbit anti-Ac-α-tub (alpha Tubulin (acetyl K40); 1:1000, Abcam, Cat# ab179484, RRID: AB_2890906).

Techniques: Expressing, Co-Culture Assay, In Vitro, Cell Culture, Labeling, Western Blot, Immunofluorescence, Two Tailed Test, Fluorescence

Journal: Neural Regeneration Research

Article Title: ATAT1 deficiency enhances microglia/macrophage-mediated erythrophagocytosis and hematoma absorption following intracerebral hemorrhage

doi: 10.4103/1673-5374.382984

Figure Lengend Snippet: Enhanced microglia/macrophage erythrophagocytosis after ATAT1 silencing in vitro . (A, B) Representative fluorescence images (A) and quantification (B) of GFP-positive RAW264.7 cells in each group (scale bar: 10 µm). (C, D) Representative fluorescence images (C) and quantification (D) of GFP-positive BV2 cells in each group (scale bar: 10 µm). (E, F) Representative flow cytometry (E) and quantification (F) of GFP-positive RAW264.7 cells in each group. (G, H) Representative flow cytometry plots (G) and quantification (H) of GFP-positive BV2 cells in each group. Data are shown as the mean ± SEM ( n = 5 independent cell cultures). * P < 0.05, ** P < 0.01, *** P < 0.001, vs . scrambled shRNA (two-tailed Student’s t -test). ATAT1: α-Tubulin acetyltransferase 1; GFP: green fluorescent protein.

Article Snippet: After blocking in 3% BSA for 2 hours, the membranes were incubated with the following primary antibodies overnight at 4°C: mouse anti-α-tubulin (1:1000, Abcam, Cambridge, UK, Cat# ab7291, RRID: AB_2241126) and rabbit anti-Ac-α-tub (alpha Tubulin (acetyl K40); 1:1000, Abcam, Cat# ab179484, RRID: AB_2890906).

Techniques: In Vitro, Fluorescence, Flow Cytometry, shRNA, Two Tailed Test

Journal: Neural Regeneration Research

Article Title: ATAT1 deficiency enhances microglia/macrophage-mediated erythrophagocytosis and hematoma absorption following intracerebral hemorrhage

doi: 10.4103/1673-5374.382984

Figure Lengend Snippet: ATAT1 deficiency enhances the phagocytic ability of microglia/macrophages after ICH in mice in vivo . (A) Representative immunofluorescence staining of areas around the hematoma in each group at 3 days after ICH. Peri-hematomal brain sections were co-stained with Iba1 (red) and 4′,6-diamidino-2-phenylindole (blue), and RBCs were labeled with DiO-GFP (scale bar: 20 µm). (B) Quantification of the Iba1-positive cells around hematoma per mm 2 in each group. (C) Percentage of GFP and Iba1 double-positive cells in all Iba1-positive cells in each group. Data are shown as the mean ± SEM ( n = 5 animals). *** P < 0.001, vs . ICH WT (two-tailed Student’s t -test). ATAT1: α-Tubulin acetyltransferase 1; GFP: green fluorescent protein; Iba1: ionized calcium binding adapter molecule 1; ICH: intracerebral hemorrhage.

Article Snippet: After blocking in 3% BSA for 2 hours, the membranes were incubated with the following primary antibodies overnight at 4°C: mouse anti-α-tubulin (1:1000, Abcam, Cambridge, UK, Cat# ab7291, RRID: AB_2241126) and rabbit anti-Ac-α-tub (alpha Tubulin (acetyl K40); 1:1000, Abcam, Cat# ab179484, RRID: AB_2890906).

Techniques: In Vivo, Immunofluorescence, Staining, Labeling, Two Tailed Test, Binding Assay

Journal: Neural Regeneration Research

Article Title: ATAT1 deficiency enhances microglia/macrophage-mediated erythrophagocytosis and hematoma absorption following intracerebral hemorrhage

doi: 10.4103/1673-5374.382984

Figure Lengend Snippet: ATAT1 deficiency accelerates hematoma absorption after ICH. (A) Representative images of the hematoma on days 3 and 7 after ICH in each group (scale bar: 1 mm). (B) Quantification of the hematoma volume (mm 3 ) on days 3 and 7 after ICH in each group. (C) Representative images of Perls staining on days 3 and 7 after ICH in each group (scale bar: 50 µm). (D) Quantification of the Perls-positive cells per mm 2 around the hematoma on days 3 and 7 after ICH in each group. Data are shown as the mean ± SEM ( n = 5 animals). * P < 0.05, ** P < 0. 01, *** P < 0.001, vs . ICH WT (two-tailed Student’s t -test). ATAT1: α-Tubulin acetyltransferase 1; ICH: intracerebral hemorrhage.

Article Snippet: After blocking in 3% BSA for 2 hours, the membranes were incubated with the following primary antibodies overnight at 4°C: mouse anti-α-tubulin (1:1000, Abcam, Cambridge, UK, Cat# ab7291, RRID: AB_2241126) and rabbit anti-Ac-α-tub (alpha Tubulin (acetyl K40); 1:1000, Abcam, Cat# ab179484, RRID: AB_2890906).

Techniques: Staining, Two Tailed Test

Journal: Neural Regeneration Research

Article Title: ATAT1 deficiency enhances microglia/macrophage-mediated erythrophagocytosis and hematoma absorption following intracerebral hemorrhage

doi: 10.4103/1673-5374.382984

Figure Lengend Snippet: ATAT1 deficiency decreases the inflammatory response after ICH (quantitative reverse transcription-polymerase chain reaction). (A–C) Quantification of the pro-inflammatory genes interleukin (IL)-1β (A), IL-6 (B), and tumor necrosis factor α (TNFα; C) in brain tissues around hematoma on day 7 after ICH in each group. (D–F) Quantification of the anti-inflammatory genes IL-4 (D), IL-10 (F), and transforming growth factor-β (TGFβ; F) in brain tissues around hematoma on day 7 after ICH in each group. Data are shown as the mean ± SEM ( n = 5 animals). * P < 0.05, *** P < 0.001, vs . ICH WT (two-tailed Student’s t -test). ATAT1: α-Tubulin acetyltransferase 1; ICH: intracerebral hemorrhage.

Article Snippet: After blocking in 3% BSA for 2 hours, the membranes were incubated with the following primary antibodies overnight at 4°C: mouse anti-α-tubulin (1:1000, Abcam, Cambridge, UK, Cat# ab7291, RRID: AB_2241126) and rabbit anti-Ac-α-tub (alpha Tubulin (acetyl K40); 1:1000, Abcam, Cat# ab179484, RRID: AB_2890906).

Techniques: Reverse Transcription Polymerase Chain Reaction, Two Tailed Test

Journal: Neural Regeneration Research

Article Title: ATAT1 deficiency enhances microglia/macrophage-mediated erythrophagocytosis and hematoma absorption following intracerebral hemorrhage

doi: 10.4103/1673-5374.382984

Figure Lengend Snippet: ATAT1 deficiency reduces neuronal death and promotes behavioral function recovery after ICH. (A) Representative immunofluorescence staining of areas around the hematoma in each group on day 7 after ICH. Peri-hematomal brain sections were co-stained with TdT-mediated dUTP nick-end labeling (TUNEL; red), neuronal nuclei (NeuN; green), and 4′,6-diamidino-2-phenylindole (DAPI; blue). Scale bar: 20 µm. (B) Quantification of TUNEL-positive neurons around the hematoma per mm 2 in each group. (C) Quantification of the neurological score in each group on days 1, 3, 7, and 14 after ICH. (D) Quantification of latency to fall (seconds) in the accelerated rotarod test in each group on days 1, 3, 7, and 14 after ICH. (E) Quantification of the contralateral limbs slip ratio (%) within 50 steps in the beam walking test in each group on days 1, 3, 7, and 14 after ICH. (F) Percentage of the right forelimb used (%) in the cylinder test in each group on days 1, 3, 7, and 14 after ICH. (R–L)/total: right forelimb usage count−left forelimb usage count)/total forelimb usage count. Data are expressed as mean ± SEM ( n = 5 animals for each group). * P < 0.05, ** P < 0.01, vs . ICH WT . Two-tailed Student’s t -test (B) and two-way analysis of variance followed by Tukey’s post hoc test (D–F) were used for data comparison. The neurological score evaluation was analyzed by a generalized linear model with generalized estimating equations (C). ATAT1: α-Tubulin acetyltransferase 1; ICH: intracerebral hemorrhage.

Article Snippet: After blocking in 3% BSA for 2 hours, the membranes were incubated with the following primary antibodies overnight at 4°C: mouse anti-α-tubulin (1:1000, Abcam, Cambridge, UK, Cat# ab7291, RRID: AB_2241126) and rabbit anti-Ac-α-tub (alpha Tubulin (acetyl K40); 1:1000, Abcam, Cat# ab179484, RRID: AB_2890906).

Techniques: Immunofluorescence, Staining, End Labeling, TUNEL Assay, Two Tailed Test, Comparison

Journal: Science Advances

Article Title: Characterizing smoking-induced transcriptional heterogeneity in the human bronchial epithelium at single-cell resolution

doi: 10.1126/sciadv.aaw3413

Figure Lengend Snippet: ( A ) Expression of gene sets GS-2, GS-3, and GS-7 in clusters C-5 and C-11 was visualized by heatmap ( z -normalized TPM values). ( B ) Cluster C-5 was split into never and current smoker subsets, and expression of GS-8 genes was visualized by heatmap. ( C ) Bronchial tissue procured from an independent cohort of never and current smokers (UMCG cohort, table S2) was immunostained for AKR1B10, Ac-α-Tub, and KRT8. Representative images of never smoker (left) and current smoker (right) tissue were displayed. Arrows specify examples of AKR1B10 + ciliated cells (Ac-α-Tub + ). ( D ) An increase in tissue length (μm)–normalized numbers of AKR1B10 + Ac-α-Tub + cells was observed in current smokers relative to never smokers [ P = 7.4 × 10 −7 , Wilcoxon rank-sum (WRS) test].

Article Snippet: Immunostaining was performed using the following primary antibodies: mouse anti–Ac-α-Tub (Sigma, T6793), rabbit anti–Ac-α-Tub (Enzo Life Sciences, BML SA4592), rabbit anti-AKR1B10 (Sigma, HPA020280), rabbit anti-CEACAM5 (Abcam, ab131070), chicken anti-KRT5 (BioLegend, 905-901), rat anti-KRT8 (Developmental Studies Hybridoma Bank, University of Iowa; TROMA-I), mouse anti-MUC5AC (Abcam, ab3649), and rabbit anti-MUC5B (Sigma, HPA008246).

Techniques: Expressing