Journal: Cell death & disease

Article Title: POU6F1 cooperates with RORA to suppress the proliferation of lung adenocarcinoma by downregulation HIF1A signaling pathway.

doi: 10.1038/s41419-022-04857-y

Figure Lengend Snippet: Fig. 5 POU6F1 inactivates HIF1A signaling pathway and transcriptionally regulates ENO1, PDK1, and PRKCB expression. A Volcano plot showing differential expression genes (DEGs) in A549 cells stably transfected with POU6F1 compared with empty vector (mock). B Human KEGG pathway analysis of DEGs indicating participated pathways in A549 cells stably transfected with POU6F1 relative to mock. C Venn diagram revealing comprehensive analysis of genes, that were involved with the HIF1A signaling pathway and associated with LUAD progression in various status of death, tumor stage, and metastasis. D Western blotting assay showing the expression of HIF1A, ENO1, ENO2, GAPDH, PDK1, and PRKCB in A549 and NCI-H1299 cells stably transfected with mock or POU6F1. E Real-time qRT-PCR assay indicating the expression of HIF1A, ENO1, PDK1, and PRKCB in A549 cells stably transfected with mock or POU6F1. F Dual-luciferase assay indicating relative activity of HIF1A promoter in A549 and NCI-H1299 cells stably transfected with mock or POU6F1, and those treated with DMSO or ML228 (1.0 μmol/l). G Western blotting assay showing the expression of HIF1A in A549 and NCI-H1299 cells stably transfected with mock or POU6F1, and those treated with DMSO or MG132 (5 μmol/l) for 6 h. H Dual-luciferase assay showing relative activity of ENO1, PDK1, and PRKCB promoter in A549 and NCI-H1299 cells transfected with mock or POU6F1. I ChIP and qPCR assays indicating relative POU6F1 enrichment on ENO1, PDK1, and PRKCB promoter in A549 cells stably transfected with mock or POU6F1. Student t-test and ANOVA compared the difference in E, F and H, I. *P < 0.05, **P < 0.01.

Article Snippet: Thereafter, membranes were blocked with 5% skimmed milk powder and incubated with the primary antibodies specific for POU6F1 (abclonal, A7299), RORA (Proteintech, 10616-1-AP; Santa Cruz Biotechnology, sc-518081), HIF1A (Abcam, ab51608), actin (Abclonal, AC026), ENO1 (Proteintech, 11204-1-AP), ENO2 (Proteintech, 66150-1-Ig), PDK1 (Proteintech, 10026-1-AP), PRKCB (Proteintech, 12919-1-AP), HA (Abcam, ab9110), Histone-H3 (Proteintech, 17168-1-AP), Flag antibody (Abcam, ab205606), and GAPDH (Abcam, ab8245) at 4 °C overnight.

Techniques: Expressing, Quantitative Proteomics, Stable Transfection, Transfection, Plasmid Preparation, Western Blot, Quantitative RT-PCR, Luciferase, Activity Assay

Journal: Brain : a journal of neurology

Article Title: Connexin43 mimetic peptide reduces vascular leak and retinal ganglion cell death following retinal ischaemia.

doi: 10.1093/brain/awr338

Figure Lengend Snippet: Figure 3 Three hours of hypoxia and 6 h of reperfusion lead to significant endothelial cell death in vitro in rat brain microvascular endothelial cells (RBMVEC; A). Non-specific gap junction blocker carbenoxolone, non-specific hemichannel blocker LaCl3, and connexin43 (Cx43) mimetic peptide protected endothelial cells against hypoxic injury, with the number of viable cells significantly higher than no treatment, while scrambled peptide did not have any protective effects. The number of viable cells was expressed as percentage of the control without hypoxia. Hypoxia and reperfusion also lead to significant propidium iodide dye uptake into primary rat brain microvascular endothelial cells indicating open hemichannels (B). Carbenoxolone, LaCl3 and connexin43 mimetic peptide significantly prevented dye uptake compared to no treatment, indicating hemichannel closure, while scrambled peptide did not have any effect. Stars denote statistical significance when compared to the control group or compared between groups in brackets; P 5 0.05.

Article Snippet: Rat brain microvascular endothelial cells (R840K-05a, Cell Applications) were plated into 24-well plates (1 105 cells/well) in rat brain microvascular endothelial cell growth media (R819K-500, Cell Applications) and allowed to settle for 16 h. Medium was then removed and replaced with Dulbecco’s Modified Eagle’s Medium/F12 containing 0.5% foetal bovine serum and 1% glutamine.

Techniques: In Vitro, Control

Journal: PLoS ONE

Article Title: Bilateral Nerve Alterations in a Unilateral Experimental Neurotrophic Keratopathy Model: A Lateral Conjunctival Approach for Trigeminal Axotomy

doi: 10.1371/journal.pone.0070908

Figure Lengend Snippet: Representative histological photographs of corneal nerves stained with anti-βIII tubulin FITC-conjugated antibody. Normal subbasal (A) and stromal (E) nerve plexus. Subbasal nerve plexus completely disappeared (B; day 1, C; day 7, D; day 14) and stromal nerve definitely decreased from day1 (F; day 1, G; day 7, H; day 14) and after trigeminal axotomy. The nerve of the contralateral eye decreased from day 1 (J; day 1, K; day 7, L; day 14).

Article Snippet: Corneas were then stained with monoclonal NL637-conjugated anti-β-III tubulin antibody (anti-Neuron-specific β-III Tubulin-NL637, R&D systems Inc. Minneapolis, MN; dilution of 1∶100) at 4C degree overnight.

Techniques: Staining

Journal: PLoS ONE

Article Title: Bilateral Nerve Alterations in a Unilateral Experimental Neurotrophic Keratopathy Model: A Lateral Conjunctival Approach for Trigeminal Axotomy

doi: 10.1371/journal.pone.0070908

Figure Lengend Snippet: Representative histological photographs of corneal nerves stained with anti-βIII tubulin FITC-conjugated antibody. Normal subbasal (A) and stromal (E) nerve plexus.Subbasal nerve plexus completely disappeared (B; day 1, C; day 7, D; day 14) (A) and stromal nerve definitely decreased (B) from day 1 (F; day 1, G; day 7, H; day 14) after trigeminal axotomy. The nerve of the contralateral eye did not change (J; day 1, K; day 7, L; day 14).

Article Snippet: Corneas were then stained with monoclonal NL637-conjugated anti-β-III tubulin antibody (anti-Neuron-specific β-III Tubulin-NL637, R&D systems Inc. Minneapolis, MN; dilution of 1∶100) at 4C degree overnight.

Techniques: Staining