Journal: Genes & Diseases

Article Title: Druggable target ATAD2 enhances the malignant progression and cooperates with E2F1 to up-regulate PDK1 expression in glioma

doi: 10.1016/j.gendis.2025.101810

Figure Lengend Snippet: The expression of the druggable target ATAD2 in glioma. (A) Visualization of DepMap database-based druggability analysis of CTARS-related proteins using a network Venn diagram. (B) Analysis using the GEPIA database analysis revealed significant upregulation of ATAD2 mRNA expression in glioblastoma (GBM). (C) The UALCAN database analysis showed significant up-regulation of ATAD2 protein in GBM. (D) The expression of ATAD2 mRNA across different clinicopathological characteristics was analyzed in the CGGA cohort. Statistical analyses utilized the Kruskal–Wallis H test followed by post hoc Dunn's test for WHO grade comparisons and the Wilcoxon rank-sum test for other parameters. (E) Representative cases showing immunohistochemical staining across various clinicopathological tissue types of gliomas. Scale bar: 100 μm. (F) Statistical analysis of the IHC scores based on the staining intensity and the percentage of positive cells. Statistical analysis was performed with the Kruskal–Wallis H test and by post hoc Dunn's test. Significance levels are indicated as follows: ns, not significant; ∗, P < 0.05; ∗∗∗∗, P < 0.0001.

Article Snippet: The cells were incubated with an ATAD2 primary antibody (BOSTER, cat# M03855, 1:200), washed with PBS containing 0.1% Tween 20, followed by incubation with a secondary antibody (CoraLite594-Goat Anti-Rabbit, Proteintech, cat# SA00013-4, 1:300).

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Genes & Diseases

Article Title: Druggable target ATAD2 enhances the malignant progression and cooperates with E2F1 to up-regulate PDK1 expression in glioma

doi: 10.1016/j.gendis.2025.101810

Figure Lengend Snippet: The impact of ATAD2 on the malignant phenotype of glioma cells. (A) The Western blot analysis shows the differential expression of ATAD2 in a normal astrocyte cell line (HA1800) and various glioma cell lines. (B) The Western blot analysis demonstrates the efficacy of three ATAD2 shRNAs in reducing ATAD2 expression in the LN229 and U251MG cell lines. (C) The Western blot analysis revealed the expression levels of ATAD2 and the FLAG-tagged protein following ATAD2 overexpression in U118MG cells. (D – F) The CCK-8 assay shows changes in cell proliferation after knockdown of ATAD2 in LN229 and U251MG cells, and its overexpression in U118MG cells. (G, H) Colony formation assay and statistical analysis. (I, J) Migration and invasion assays and their statistical analysis. The data are presented as means ± SD. For panels (D – F), two-way ANOVA was performed, while unpaired two-tailed Student's t -tests were used for panels (H, I). Statistical significance is denoted as follows: ∗, P < 0.05; ∗∗, P < 0.01; ∗∗∗, P < 0.001; ∗∗∗∗, P < 0.0001.

Article Snippet: The cells were incubated with an ATAD2 primary antibody (BOSTER, cat# M03855, 1:200), washed with PBS containing 0.1% Tween 20, followed by incubation with a secondary antibody (CoraLite594-Goat Anti-Rabbit, Proteintech, cat# SA00013-4, 1:300).

Techniques: Western Blot, Quantitative Proteomics, Expressing, Over Expression, CCK-8 Assay, Knockdown, Colony Assay, Migration, Two Tailed Test

Journal: Genes & Diseases

Article Title: Druggable target ATAD2 enhances the malignant progression and cooperates with E2F1 to up-regulate PDK1 expression in glioma

doi: 10.1016/j.gendis.2025.101810

Figure Lengend Snippet: The impact of ATAD2 knockdown on Glioma tumorigenesis in vivo . (A) Gross morphological observations were conducted on subcutaneous tumors induced by LN229 cells in the Shco and Sh-3 groups of nude mice. (B) Statistical analysis of the subcutaneous tumor size measurements. (C) Statistical analysis of the subcutaneous xenograft tumor weights. (D) Representative images of IHC staining for ATAD2 and Ki67. Scale bar:100 μm. (E, F) The Image-Pro Plus software (version 6) was used to compute the sum integrated optical density of ATAD2 and Ki-67 IHC staining, followed by statistical analysis. (G) Representative HE staining images of intracranial orthotopic xenografts formed by LN229 cells in the Shco and Sh-3 groups. Scale bar: 1 mm. (H) Statistical analysis of intracranial orthotopic tumor size measurements. (I) Kaplan–Meier survival curve of nude mice with orthotopic intracranial tumors (Log-rank test). The data are presented as means ± SD, two-tailed paired Student's t -test was used to compare the results in (B), (C), (E), and (F); and two-tailed unpaired Student's t -test was used to compare the results in (H). Each group included n = 6 mice. Statistical significance is denoted as follows: ∗∗∗, P < 0.001; ∗∗∗∗, P < 0.0001.

Article Snippet: The cells were incubated with an ATAD2 primary antibody (BOSTER, cat# M03855, 1:200), washed with PBS containing 0.1% Tween 20, followed by incubation with a secondary antibody (CoraLite594-Goat Anti-Rabbit, Proteintech, cat# SA00013-4, 1:300).

Techniques: Knockdown, In Vivo, Immunohistochemistry, Software, Staining, Two Tailed Test

Journal: Genes & Diseases

Article Title: Druggable target ATAD2 enhances the malignant progression and cooperates with E2F1 to up-regulate PDK1 expression in glioma

doi: 10.1016/j.gendis.2025.101810

Figure Lengend Snippet: ATAD2-E2F1 positive feedback loop regulates the expression of PDK1. (A) Volcano plot of differentially expressed genes from RNA-seq analysis of ATAD2 knockdown and control conditions in LN229 cells. (B) Volcano plot of differentially expressed proteins from proteome analysis of ATAD2 knockdown and control conditions in LN229 cells. (C) The Venn diagram illustrates the overlap between the up-regulated and down-regulated mRNAs and proteins. (D) The fold change ranking plot illustrates 20 commonly down-regulated proteins. (E, F) Western blot analysis confirmed that ATAD2 enhances the expression of PDK1. (G, H) Western blot analysis confirmed that E2F1 up-regulates the expression of ATAD2. (I–K) Western blot analysis validated that E2F1 enhances the expression of ATAD2. (L, M) Western blot analysis showed that ATAD2 cooperates with E2F1 to up-regulate the expression of PDK1. (N) The dual luciferase reporter gene assays revealed that ATAD2 and E2F1 synergistically enhance the promoter activity of PDK1. The data are presented as means ± SD. Statistical comparisons were performed using unpaired two-tailed Student's t -test for figures (F), (H), and (K), and one-way ANOVA followed by Tukey's post hoc test for figures (J), (M), and (N). Statistical significance is denoted as follows: ns, not significant; ∗, P < 0.05; ∗∗, P < 0.01; ∗∗∗, P < 0.001; ∗∗∗∗, P < 0.0001.

Article Snippet: The cells were incubated with an ATAD2 primary antibody (BOSTER, cat# M03855, 1:200), washed with PBS containing 0.1% Tween 20, followed by incubation with a secondary antibody (CoraLite594-Goat Anti-Rabbit, Proteintech, cat# SA00013-4, 1:300).

Techniques: Expressing, RNA Sequencing, Knockdown, Control, Western Blot, Luciferase, Activity Assay, Two Tailed Test

Journal: Genes & Diseases

Article Title: Druggable target ATAD2 enhances the malignant progression and cooperates with E2F1 to up-regulate PDK1 expression in glioma

doi: 10.1016/j.gendis.2025.101810

Figure Lengend Snippet: The clinical significance of the ATAD2-E2F1-PDK1 axis in glioma. (A, B) Analysis using the GlioVis database showed a positive correlation between the expressions of ATAD2, E2F1, and PDK1, as determined by the Spearman correlation test. (C) Representative cases showed the expression of E2F1 and PDK1 in glioma clinical specimens with low and high expression of ATAD2. Scale bar: 100 μm. (D) Spearman's correlation analysis of ATAD2, E2F1, and PDK1 expression levels in glioma clinical specimens. (E) Kaplan–Meier analysis and log-rank test of survival rates across distinct co-expression groups of ATAD2, E2F1, and PDK1 in the CGGA cohort. P values were adjusted for multiple group comparisons using the Bonferroni method. (F) Graphical diagram illustrating that ATAD2 promotes glioma progression and synergizes with E2F1 to increase PDK1 expression. The figure was created using Figdraw ( www.figdraw.com ). Statistical significance is denoted as follows: ∗∗∗, P < 0.001.

Article Snippet: The cells were incubated with an ATAD2 primary antibody (BOSTER, cat# M03855, 1:200), washed with PBS containing 0.1% Tween 20, followed by incubation with a secondary antibody (CoraLite594-Goat Anti-Rabbit, Proteintech, cat# SA00013-4, 1:300).

Techniques: Expressing

Journal: Acta biochimica et biophysica Sinica

Article Title: Apoptotic mechanism of MCF-7 breast cells in vivo and in vitro induced by photodynamic therapy with C-phycocyanin.

doi: 10.1093/abbs/gmp104

Figure Lengend Snippet: Figure 7 Western blot analysis of caspase-9 expression MCF-7 cells were analyzed by 15% SDS–PAGE, and after electrophoresis, proteins on the gel were transferred to nitrocellulose membrane and probed with mouse anti-caspase-9 monoclonal antibody. Lane 1: control group; lane 2: He–Ne laser group; lane 3: C-PC group; lane 4: C-PC þ He–Ne laser group.

Article Snippet: Then, the slides were incubated with blocking buffer followed by the biotinylated rabbit antihuman Bcl-2 monoclonal antibody (Boster, 1:100) and SABC.

Techniques: Western Blot, Expressing, SDS Page, Electrophoresis, Membrane, Control

Journal: Molecular Medicine Reports

Article Title: Effect of Melilotus suaveolens extract on pulmonary microvascular permeability by downregulating vascular endothelial growth factor expression in rats with sepsis

doi: 10.3892/mmr.2015.3146

Figure Lengend Snippet: Effect of M. suaveolens extract on the expression of VEGF and NF-κB65 protein in lung tissue. Groups of mice were challenged with cecal ligation and puncture and treated with M. suaveolens extract 24 h later. The expression of VEGF, NF-κβ65 and GAPDH was detected by western blotting using specific antibodies. GAPDH protein was used an internal control. (A) Representative western blot analysis demonstrated the levels of VEGF and NF-κB65 protein expression in rats from the four groups; (a) normal control group; (b) sham operation group; (c) (untreated) sepsis group and (d) treatment group. (B) Quantification of the blots by densitometric analysis of (a) VEGF and (b) NF-κB65 protein expression in rats from the four groups. Data are presented as the mean ± standard deviation of one experiment consisting of three replicates. ** P<0.01 vs. the sham operation group and normal control group; # P<0.05 vs . (untreated) sepsis group. VEGF, vascular endothelial growth factor; NF-κB, nuclear factor kappa B.

Article Snippet: Primary antibodies used included rabbit anti-VEGF monoclonal antibody (1:400), rabbit anti-NF-κB65 monoclonal antibody (1:400) (Boster Biological Technology, Ltd) and mouse anti- GAPDH monoclonal antibody (1:400) (Santa Cruz Biotechnology, Inc).

Techniques: Expressing, Ligation, Western Blot, Control, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: Effect of Melilotus suaveolens extract on pulmonary microvascular permeability by downregulating vascular endothelial growth factor expression in rats with sepsis

doi: 10.3892/mmr.2015.3146

Figure Lengend Snippet: Effect of M. suaveolens extract on the protein expression of VEGF and NF-κB65 in rat lungs 24 h following cecal ligation and puncture-induced acute lung injury. Immunostaining was performed on lung sections following antigen retrieval using Retrievagen. (A) Representative immunostaining revealed VEGF and NF-κβ65-positive expression in rats from the four groups: (a–c) Expression of positive VEGF in the (a) sham operation group; (b) control group; (c) treatment group); (d–f) Expression of positive NF-κB65 in the (d) sham operation group; (e) (untreated) sepsis group; (f) treatment group (magnification, ×200). (B) Quantification of the images by densitometric analysis of (a) VEGF and (b) NF-κB-positive protein expression in rats from four groups. All values are expressed as the mean ± standard deviation. ** P<0.01 vs. the sham operation group; ## P<0.01 vs. (untreated) sepsis group. VEGF, vascular endothelial growth factor; NF-κB, nuclear factor kappa B.

Article Snippet: Primary antibodies used included rabbit anti-VEGF monoclonal antibody (1:400), rabbit anti-NF-κB65 monoclonal antibody (1:400) (Boster Biological Technology, Ltd) and mouse anti- GAPDH monoclonal antibody (1:400) (Santa Cruz Biotechnology, Inc).

Techniques: Expressing, Ligation, Immunostaining, Control, Standard Deviation

Journal: Autophagy

Article Title: Mutant HTT (huntingtin) impairs mitophagy in a cellular model of Huntington disease

doi: 10.1080/15548627.2020.1728096

Figure Lengend Snippet: PolyQ tract in mutant HTT does not affect the polyUB labeling of mitochondria. (a) Representative immunoblots of total polyUB (total-UB) and phosphorylated ubiquitin (Ser65; p-UB-S65) in total homogenate and isolated mitochondria from ST-Q7 and ST-Q111 control cells (vehicle, CTR) or treated with rotenone (Rot, 1 μM, 4 h) or CCCP (10 μM, 24 h). Protein levels were normalized relative to Ponceau staining and quantification is depicted as fold-change to control ST-Q7 cells of at least three independent experiments. (b) Representative immunoblots of BNIP3 and BNIP3L in isolated mitochondria from ST-Q7 and ST-Q111 control cells (vehicle, CTR) or treated with rotenone (Rot, 1 μM, 4 h) or CCCP (10 μM, 24 h). Protein levels were quantified using all bands present in each lane independently of the molecular weight or oligomerization state (indicated with line and arrows) and were normalized relative to Ponceau staining and quantification is depicted as fold-change to control ST-Q7 cells. Data are presented as mean ± s.e.m of at least 3 independent experiments and no significant changes were observed after statistical analysis

Article Snippet: OPTN (10837-1-AP) from Proteintech Group, HTT (MAB2166) from Chemicon, SQSTM1/p62 (GP62-C) and NBR1 (H00004077-M01) from Abnova, anti-DNA (AC-30-10) from Progen, MTOR (GT649) mouse (GTX630198) from Genetex, total-UB (U5379) and ACTB/β-actin (A5441) from Sigma-Aldrich, BNIP3 (M01469), BNIP3L (M03107) and FUNDC1 (A08688) from Boster.

Techniques: Mutagenesis, Labeling, Western Blot, Ubiquitin Proteomics, Isolation, Control, Staining, Molecular Weight