Journal: Frontiers in Molecular Biosciences

Article Title: TUBB2A expression and its prognostic significance in hepatocellular carcinoma revealed by cholesterol-metabolism-related gene profiling

doi: 10.3389/fmolb.2026.1778506

Figure Lengend Snippet: Overall workflow and results of key gene screening and survival analysis. (A) Volcano plot of differentially expressed genes in GSE166163 . (B) Venn diagram showing the intersection between function-related genes from GeneCards (cholesterol metabolism, cell proliferation, and apoptosis) and upregulated genes (n = 139). (C) Cross-validation plot of the LASSO regression model used to determine the optimal λ value and select feature genes. (D) Variable importance ranking of candidate genes generated using the random forest model. (E) Twenty-three core genes obtained from the intersection of LASSO and random forest results. (F) Univariate Cox regression analysis based on TCGA-LIHC identifying nine genes significantly associated with overall survival (OS). (G–L) Kaplan-Meier survival curves of six representative genes (LOX, MMP3, MMP9, TFE3, TUBB2A, and UCHL1).

Article Snippet: The primary antibody against TUBB2A (Novus, catalog number NBP3-16484-100 μL) was diluted following the manufacturer’s guidelines and applied to tissue sections, which were incubated 2 h. Then sections were subjected to three washes using Tris-buffered saline, with each wash performed for 5 min.

Techniques: Biomarker Discovery, Generated

Journal: Frontiers in Molecular Biosciences

Article Title: TUBB2A expression and its prognostic significance in hepatocellular carcinoma revealed by cholesterol-metabolism-related gene profiling

doi: 10.3389/fmolb.2026.1778506

Figure Lengend Snippet: Expression characteristics of key genes in the GSE151530 single-cell atlas. (A) t-SNE plot displaying single-cell clusters and annotation of major cell types. (B) Feature plots showing the distribution of candidate genes (LOX, MMP3, MMP9, TFE3, TUBB2A, and UCHL1) across different cell subpopulations. (C) Violin plots illustrating differential expression of TUBB2A between HCC and normal tissues across various cell types.

Article Snippet: The primary antibody against TUBB2A (Novus, catalog number NBP3-16484-100 μL) was diluted following the manufacturer’s guidelines and applied to tissue sections, which were incubated 2 h. Then sections were subjected to three washes using Tris-buffered saline, with each wash performed for 5 min.

Techniques: Expressing, Single Cell, Quantitative Proteomics

Journal: Frontiers in Molecular Biosciences

Article Title: TUBB2A expression and its prognostic significance in hepatocellular carcinoma revealed by cholesterol-metabolism-related gene profiling

doi: 10.3389/fmolb.2026.1778506

Figure Lengend Snippet: Expression of TUBB2A in HCC tissues and its prognostic significance. (A) Above: HE staining showing tissue morphology; below: immunohistochemical staining showing TUBB2A expression in the cytoplasm of hepatocellular carcinoma tissues, with higher expression in tumor tissues compared to adjacent normal tissues. Left: adjacent normal tissue with low TUBB2A expression, middle: HCC with lower TUBB2A expression, right: HCC with higher TUBB2A expression (magnification ×200). (B) RT-qPCR analysis of TUBB2A expression in normal and HCC tissues. (C) Western blot bands and quantitative analysis of TUBB2A expression in normal and tumor tissues (N: normal; T: tumor). (D) Kaplan-Meier survival curves of HCC patients stratified by low versus high TUBB2A expression (n = 121). (E) Kaplan–Meier survival curves of HCC patients with tumor diameter ≥5 cm, stratified by TUBB2A expression level (n = 59). Note: *** P ≤ 0.001; **** P ≤ 0.0001.

Article Snippet: The primary antibody against TUBB2A (Novus, catalog number NBP3-16484-100 μL) was diluted following the manufacturer’s guidelines and applied to tissue sections, which were incubated 2 h. Then sections were subjected to three washes using Tris-buffered saline, with each wash performed for 5 min.

Techniques: Expressing, Staining, Immunohistochemical staining, Quantitative RT-PCR, Western Blot

Journal: Frontiers in Molecular Biosciences

Article Title: TUBB2A expression and its prognostic significance in hepatocellular carcinoma revealed by cholesterol-metabolism-related gene profiling

doi: 10.3389/fmolb.2026.1778506

Figure Lengend Snippet: Effects of TUBB2A knockdown on the malignant behavior of HCC cells. (A) Effects of TUBB2A knockdown on the clonogenic capacity of Hep3B cells. (B) Effects of TUBB2A knockdown on Hep3B cell viability as measured by the CCK-8 assa. (C) Effects of TUBB2A knockdown on the migratory and invasive abilities of Hep3B cells, as shown by representative images and quantitative analysis of cell numbers. (D) Representative images and quantitative analysis of scratch-wound healing in Hep3B cells following TUBB2A knockdown. Note: NC group, negative control; si group, TUBB2A knockdown. * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001; **** P ≤ 0.0001.

Article Snippet: The primary antibody against TUBB2A (Novus, catalog number NBP3-16484-100 μL) was diluted following the manufacturer’s guidelines and applied to tissue sections, which were incubated 2 h. Then sections were subjected to three washes using Tris-buffered saline, with each wash performed for 5 min.

Techniques: Knockdown, CCK-8 Assay, Negative Control

Journal: Frontiers in Molecular Biosciences

Article Title: TUBB2A expression and its prognostic significance in hepatocellular carcinoma revealed by cholesterol-metabolism-related gene profiling

doi: 10.3389/fmolb.2026.1778506

Figure Lengend Snippet: Effects of TUBB2A overexpression on the malignant phenotypes of HCC cells. (A) Effects of TUBB2A overexpression on the clonogenic capacity of LM3 cells. (B) Effects of TUBB2A overexpression on LM3 cell viability as measured by the CCK-8 assay. (C) Representative images and quantification of cell numbers showing the effects of TUBB2A overexpression on migration and invasion in LM3 cells. (D) Representative images and quantitative analysis of scratch-wound healing in LM3 cells following TUBB2A overexpression. Note: NC group, negative control group; OE group, TUBB2A overexpression group. * P ≤ 0.05; ** P ≤ 0.01. *** P ≤ 0.001; **** P ≤ 0.0001.

Article Snippet: The primary antibody against TUBB2A (Novus, catalog number NBP3-16484-100 μL) was diluted following the manufacturer’s guidelines and applied to tissue sections, which were incubated 2 h. Then sections were subjected to three washes using Tris-buffered saline, with each wash performed for 5 min.

Techniques: Over Expression, CCK-8 Assay, Migration, Negative Control

Journal: Frontiers in Molecular Biosciences

Article Title: TUBB2A expression and its prognostic significance in hepatocellular carcinoma revealed by cholesterol-metabolism-related gene profiling

doi: 10.3389/fmolb.2026.1778506

Figure Lengend Snippet: Effects of TUBB2A on the in vivo tumorigenic ability of HCC cells. (A) Representative images of subcutaneous tumors formed in nude mice in the two groups. Above: Subcutaneous tumors of the control group; below: Subcutaneous tumors of the shTUBB2A group. (B) Growth curves showing changes in subcutaneous tumor volume in the two groups of nude mice. (C) Comparison of subcutaneous tumor weight between the two groups of nude mice. (D) Above: HE staining showing tissue morphology; below: IHC Ki-67 staining. Left: TUBB2A control group nude mouse tumor with low Ki-67 expression; right: TUBB2A knockdown group nude mouse tumor with high Ki-67 expression (magnification ×200). (E) Quantification of Ki-67 IHC score in the subcutaneous tumors. Note: *** P ≤ 0.001; **** P ≤ 0.0001.

Article Snippet: The primary antibody against TUBB2A (Novus, catalog number NBP3-16484-100 μL) was diluted following the manufacturer’s guidelines and applied to tissue sections, which were incubated 2 h. Then sections were subjected to three washes using Tris-buffered saline, with each wash performed for 5 min.

Techniques: In Vivo, Control, Comparison, Staining, Expressing, Knockdown

Journal: bioRxiv

Article Title: Acentrosomal spindle assembly and stability in C. elegans oocytes requires a kinesin-12 non-motor microtubule interaction domain

doi: 10.1101/2021.06.17.448874

Figure Lengend Snippet: a) MBP-MESP-1 direct interaction with N-stalk or C-stalk was tested by MBP pulldown. Reactions using MBP-MESP-1 or MBP as bait are shown. KLP-18 truncations visualized on Western blot with anti-His antibody; MBP and MBP-MESP-1 visualized by Coomassie stain. b) Microtubule co-sedimentation assay for MBP-MESP-1 with no microtubules added (-), undigested (MT), and subtilisin-digested (sMT) microtubules. Representative blots show supernatant (S) and pellet (P) samples, quantification is of average shift +/- sd. n = 2 experiments for non-subtilisin experiment, n = 3 experiments for subtilisin experiment. c) Schematic of GFP tagged proteins used in (d) and (e). d) KLP-18 recruitment to microtubules was tested by visualization of GFP tagged stalk proteins on TMR-microtubules. Representative images with (+) and without (-) MBP-MESP-1 added to the indicated GFP-tagged proteins are shown for each condition. Scale bar = 5μm. e) Quantification of mean GFP intensity overlayed on the microtubule normalized to mean tubulin intensity. Bars represent mean percentage +/- sd for each experiment over n = 3 experiments.

Article Snippet: First, GMPCPP seeds were nucleated by incubating 13µL PEM104 (104mM PIPES pH 6.9, 1.3mM EGTA, 6.3mM MgCl 2 ), 2.2µL 10mM GMPCPP, 2.2µL DMSO, 4µL 13mg/mL homemade porcine brain tubulin (gift of Sarah Rice), and 4µL 10mg/mL TMR-tubulin (Cytoskeleton) at 37°C for 40min.

Techniques: Western Blot, Staining, Sedimentation