Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Super enhancer-driven LINC01013 mediates hypoxia-induced mitochondrial dysfunction by HSPA9 to determine pulmonary arterial smooth muscle cell fate

doi: 10.1007/s00018-025-06071-3

Figure Lengend Snippet: Silencing of LINC01013 inhibited hypoxia-induced hPASMC proliferation and inflammation. A , B hPASMC proliferation was determined by CCK8 and EdU incorporation assays ( n = 6). EdU (red), DAPI (blue), scale bar, 50 μm. C Western blotting analysis of PCNA, cyclin A and cyclin D in hPASMCs ( n = 6). D RT‒qPCR analysis showed the mRNA levels of TNF-α and IL-6 after LINC01013 knockdown, β-actin was used as a internal reference gene ( n = 6). E Western blotting analysis of TNF-α and IL-6 in hPASMCs. ( n = 6). All values are presented as the mean ± SEM. Statistical analysis was performed with one-way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001. Nor, normoxia; Hyp, hypoxia; NC, negative control; si, siRNA

Article Snippet: The antibody against CEBPB (PB9171, BA0670, 1:500, Boster, Wuhan, China), H3K27ac (A7253, 1:500, ABclonal, Wuhan, China), H3K4me1 (A2355, 1:500, Wuhan, China), PCNA (A00125, 1:500, Boster, Wuhan, China), Cyclin A (PB0515, 1:500, Boster, Wuhan, China), Cyclin D (BM4272, 1:500, Boster, Wuhan, China), IL-6 (AF7236, 1:500, Beyotime, Shanghai, China), TNF-α (AF8208, 1:500, Beyotime, Shanghai, China), PKM2 (4053, 1:1000, Cell Signaling, MA, US), HK II (66974-1-Ig, 1:1000, Proteintech, IL, USA), PDH (2784, 1:1000, Cell Signaling, MA, US), HSPA9 (14887-1-AP, 1:5000, Proteintech, IL, USA), VDAC1 (10866-1-AP, 1:5000, Proteintech, IL, USA), and β-actin (TA-09, 1:1000, ZSGB‐BIO, Beijing, China) was incubated at 4 °C overnight, followed by incubation with appropriate horseradish peroxidase-conjugated secondary antibodies at room temperature for 1 h, and proteins were visualized with enhanced chemiluminescence reagents.

Techniques: Western Blot, Knockdown, Negative Control

Journal: Cancer Biology & Therapy

Article Title: Interaction between ZMIZ2 and AR promotes prostate cancer proliferation in vitro and in vivo

doi: 10.1080/15384047.2025.2604936

Figure Lengend Snippet: To uncover the molecular regulatory network underlying the functions of ZMIZ2 and AR, RNA-seq analysis was meticulously performed to identify the common downstream target genes of these two factors. (a) RNA-seq analysis of differentially expressed genes after ZMIZ2 silencing or AR silencing. (b) Venn diagram analysis of genes commonly upregulated by ZMIZ2 and AR. (c) KEGG analysis of genes commonly upregulated by ZMIZ2 and AR. (d) GO analysis of genes commonly upregulated by ZMIZ2 and AR. (e) Heatmap of the expression levels of cell cycle-related genes after ZMIZ2 silencing or AR silencing. (f–g) A flow cytometry assay was employed to determine the cell cycle distribution. (h) QPCR was utilized to assess the mRNA transcriptional levels of cell cycle-related genes. (i) Western blot analysis was carried out to detect the protein expression levels of CDK1, CCNA2, and CCNE2 in each sample group. Significant differences are indicated as follows: * p < 0.05, ** p < 0.01, and *** p < 0.001; ns indicates not significant; n = 3.

Article Snippet: The ZMIZ2 antibody (Novus Biologicals, LLC, Centennial, CO, USA), AR antibody (Santa Cruz Biotechnology, Dallas, TX, USA), CDK1 antibody (Santa Cruz Biotechnology, Dallas, TX, USA), CCNA2 antibody (Santa Cruz Biotechnology, Dallas, TX, USA), CCNE2 antibody (Santa Cruz Biotechnology, Dallas, TX, USA) were added and incubated overnight at 4 °C and then incubated with HRP-conjugated goat anti-rabbit IgG (Thermo Fisher Scientific, Waltham, MA, USA) at 37 °C for 30 min.

Techniques: RNA Sequencing, Expressing, Flow Cytometry, Western Blot

Journal: Cancer Biology & Therapy

Article Title: Interaction between ZMIZ2 and AR promotes prostate cancer proliferation in vitro and in vivo

doi: 10.1080/15384047.2025.2604936

Figure Lengend Snippet: Depletion of ZMIZ2 expression is associated with reduced AR enrichment on the promoters of downstream target genes, accompanied by a concurrent decrease in H3K27ac levels. (a) Flow chart of the ChIP experiment. (b) The binding sites of AR on the promoters of CDK1, CCNA2, and CCNE2. (c–h) ChIP analysis of AR enrichment on the CDK1, CCNA2, and CCNE2 promoters and H3K27ac levels. Significant differences are indicated as follows: * p < 0.05, ** p < 0.01, and *** p < 0.001; ns indicates not significant; n = 3.

Article Snippet: The ZMIZ2 antibody (Novus Biologicals, LLC, Centennial, CO, USA), AR antibody (Santa Cruz Biotechnology, Dallas, TX, USA), CDK1 antibody (Santa Cruz Biotechnology, Dallas, TX, USA), CCNA2 antibody (Santa Cruz Biotechnology, Dallas, TX, USA), CCNE2 antibody (Santa Cruz Biotechnology, Dallas, TX, USA) were added and incubated overnight at 4 °C and then incubated with HRP-conjugated goat anti-rabbit IgG (Thermo Fisher Scientific, Waltham, MA, USA) at 37 °C for 30 min.

Techniques: Expressing, Binding Assay

Journal: Cancer Biology & Therapy

Article Title: Interaction between ZMIZ2 and AR promotes prostate cancer proliferation in vitro and in vivo

doi: 10.1080/15384047.2025.2604936

Figure Lengend Snippet: To uncover the molecular regulatory network underlying the functions of ZMIZ2 and AR, RNA-seq analysis was meticulously performed to identify the common downstream target genes of these two factors. (a) RNA-seq analysis of differentially expressed genes after ZMIZ2 silencing or AR silencing. (b) Venn diagram analysis of genes commonly upregulated by ZMIZ2 and AR. (c) KEGG analysis of genes commonly upregulated by ZMIZ2 and AR. (d) GO analysis of genes commonly upregulated by ZMIZ2 and AR. (e) Heatmap of the expression levels of cell cycle-related genes after ZMIZ2 silencing or AR silencing. (f–g) A flow cytometry assay was employed to determine the cell cycle distribution. (h) QPCR was utilized to assess the mRNA transcriptional levels of cell cycle-related genes. (i) Western blot analysis was carried out to detect the protein expression levels of CDK1, CCNA2, and CCNE2 in each sample group. Significant differences are indicated as follows: * p < 0.05, ** p < 0.01, and *** p < 0.001; ns indicates not significant; n = 3.

Article Snippet: After electrophoresis, protein was transferred to PVDF and sealed with 5% milk for 1 h. Add following primary antibodies and incubate overnight: ZMIZ2 (Novus Biologicals, LLC, Centennial, CO, USA), AR (Santa Cruz Biotechnology, Dallas, TX, USA), CDK1 (Santa Cruz Biotechnology, Dallas, TX, USA), CCNA2 (Santa Cruz Biotechnology, Dallas, TX, USA), CCNE2 (Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: RNA Sequencing, Expressing, Flow Cytometry, Western Blot

Journal: Cancer Biology & Therapy

Article Title: Interaction between ZMIZ2 and AR promotes prostate cancer proliferation in vitro and in vivo

doi: 10.1080/15384047.2025.2604936

Figure Lengend Snippet: Depletion of ZMIZ2 expression is associated with reduced AR enrichment on the promoters of downstream target genes, accompanied by a concurrent decrease in H3K27ac levels. (a) Flow chart of the ChIP experiment. (b) The binding sites of AR on the promoters of CDK1, CCNA2, and CCNE2. (c–h) ChIP analysis of AR enrichment on the CDK1, CCNA2, and CCNE2 promoters and H3K27ac levels. Significant differences are indicated as follows: * p < 0.05, ** p < 0.01, and *** p < 0.001; ns indicates not significant; n = 3.

Article Snippet: After electrophoresis, protein was transferred to PVDF and sealed with 5% milk for 1 h. Add following primary antibodies and incubate overnight: ZMIZ2 (Novus Biologicals, LLC, Centennial, CO, USA), AR (Santa Cruz Biotechnology, Dallas, TX, USA), CDK1 (Santa Cruz Biotechnology, Dallas, TX, USA), CCNA2 (Santa Cruz Biotechnology, Dallas, TX, USA), CCNE2 (Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Expressing, Binding Assay