Journal: Cell reports

Article Title: NIX initiates mitochondrial fragmentation via DRP1 to drive epidermal differentiation

doi: 10.1016/j.celrep.2021.108689

Figure Lengend Snippet:

Article Snippet: Membranes were transferred into TrueBlack WB blocking buffer (Biotium #23013) for 60 min at room temperature then were probed with primary antibodies diluted in TrueBlack WB antibody diluent (Biotium #23013) plus 0.2% Tween-20 for 18 hr at 4°C.

Techniques: Virus, Recombinant, Electron Microscopy, Transfection, Blocking Assay, Plasmid Preparation, CRISPR, Sequencing, Software

Journal: Cell Death & Disease

Article Title: LINC01106 post-transcriptionally regulates ELK3 and HOXD8 to promote bladder cancer progression

doi: 10.1038/s41419-020-03236-9

Figure Lengend Snippet: a The relative expression of LINC01116 was analyzed in patients with different stages of BCa tissues using the GEPIA 2 bioinformatics website. b The overall survival rate was detected by GEPIA 2 bioinformatics website. c RT-qPCR was used to detect the expression of LINC01116 in human bladder epithelial immortalized cell line SV-HUC-1 and human BCa cell lines (RT-4, 5637, J82, and T24). d RT-qPCR analysis detected LINC01116 expression in J82 and T24 cells transfected with shRNAs targeting LINC01116. e , f Colony formation assay, and EdU assay tested whether LINC01116 knockdown affects the proliferation ability of BCa cell lines. g , h JC-1 assay, and flow cytometry analysis revealed the apoptosis rate of J28 and T24 cells transfected with shRNAs targeting LINC01116. i , j Tumor volume, and tumor weight were measured in response to LINC01116 knockdown. k IHC assay detected the staining of Ki67 and PCNA in tumors with or without LINC01116 knockdown. * P < 0.05, ** P < 0.01.

Article Snippet: The fixed J82 and T24 cells were rinsed in PBS, dehydrated, and air-dried for cultivating in hybridization buffer with LINC01116-specific RNA fluorescence in situ hybridization (FISH) probe (Ribobio).

Techniques: Expressing, Quantitative RT-PCR, Transfection, Colony Assay, EdU Assay, Flow Cytometry, Staining

Journal: Cell Death & Disease

Article Title: LINC01106 post-transcriptionally regulates ELK3 and HOXD8 to promote bladder cancer progression

doi: 10.1038/s41419-020-03236-9

Figure Lengend Snippet: a , b Transwell assays were performed to explore the role of LINC01116 knockdown in BCa cells migration and invasion. c E-cadherin, Vimentin, and N-cadherin expression were measured by western blot in J82 and T24 cells transfected with sh-LINC01116#1 and sh-LINC01116#2. d IF assay determined the staining of E-cadherin and N-cadherin in J82 and T24 cells with or without LINC01116 silence. e The representative image of lungs from the two groups, and HE staining of metastasis nodules in these lungs. * P < 0.05, ** P < 0.01.

Article Snippet: The fixed J82 and T24 cells were rinsed in PBS, dehydrated, and air-dried for cultivating in hybridization buffer with LINC01116-specific RNA fluorescence in situ hybridization (FISH) probe (Ribobio).

Techniques: Migration, Expressing, Western Blot, Transfection, Staining

Journal: Cell Death & Disease

Article Title: LINC01106 post-transcriptionally regulates ELK3 and HOXD8 to promote bladder cancer progression

doi: 10.1038/s41419-020-03236-9

Figure Lengend Snippet: a , b . Nuclear separation followed by RT-qPCR and AGE, as well as FISH assay, monitored the subcellular location of LINC01116 in BCa cells. c , d GEPIA 2 analyzed the relationship between two protein-coding genes (ELK3 and HOXD8) and LINC01116 in BLCA tumors. e , f RT-qPCR and western blot analyzed the expression of ELK3 in BCa cells under LINC01116 inhibition. g , h RT-qPCR and western blot analyzed the expression of HOXD8 in BCa cells. i , j Dual-luciferase reporter assay further validated the combination between ELK3/HOXD8 and LINC01116. * P < 0.05, ** P < 0.01.

Article Snippet: The fixed J82 and T24 cells were rinsed in PBS, dehydrated, and air-dried for cultivating in hybridization buffer with LINC01116-specific RNA fluorescence in situ hybridization (FISH) probe (Ribobio).

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Inhibition, Luciferase, Reporter Assay

Journal: Cell Death & Disease

Article Title: LINC01106 post-transcriptionally regulates ELK3 and HOXD8 to promote bladder cancer progression

doi: 10.1038/s41419-020-03236-9

Figure Lengend Snippet: a Ago2-RIP assay was performed to prove the existence of LINC01116 and ELK3 in RISCs. b , c By means of searching on starBase, miR-3612 was predicted to have the potentials to bind with LINC01116 andELK3. d , e Luciferase reporter gene assays predicted the relationship between miR-3612 and LINC01116/ELK3. f RNA pull-down assay further examined the combination between miR-3612 and LINC01116/ELK3. g Ago2-RIP assay confirmed the interaction among LINC01116, miR-3612, and ELK3 in RISCs. h RNA pull-down assay detected the impact of LINC01116 on the interaction of miR-3612 with ELK3. * P < 0.05, ** P < 0.01.

Article Snippet: The fixed J82 and T24 cells were rinsed in PBS, dehydrated, and air-dried for cultivating in hybridization buffer with LINC01116-specific RNA fluorescence in situ hybridization (FISH) probe (Ribobio).

Techniques: Luciferase, Pull Down Assay

Journal: Cell Death & Disease

Article Title: LINC01106 post-transcriptionally regulates ELK3 and HOXD8 to promote bladder cancer progression

doi: 10.1038/s41419-020-03236-9

Figure Lengend Snippet: a StarBase software predicted thirteen RBPs shared between HOXD8 and LINC01116. b RT-qPCR detected the relative expression of HOXD8 in J28 cells transfected with shRNAs targeting indicated RBPs. c RNA pull-down assay tested the interaction of LINC01116 with DKC1 or ELACL1. d RNA pull-down assay detected the interaction between HOXD8 and DKC1. e RIP assay verified whether LINC01116 or HOXD8 could be enriched in the anti-DKC1 pallet. f , g RT-qPCR analysis, and western blot analyzed that the expression of DKC1/HOXD8 in J82 and T24 cells transfected with shRNAs against DKC1. h RT-qPCR examined the stability of HOXD8 under ActD treatment in J82 and T24 cells transfected sh-LINC01116#1 or sh-DKC1#1. * P < 0.05, ** P < 0.01.

Article Snippet: The fixed J82 and T24 cells were rinsed in PBS, dehydrated, and air-dried for cultivating in hybridization buffer with LINC01116-specific RNA fluorescence in situ hybridization (FISH) probe (Ribobio).

Techniques: Software, Quantitative RT-PCR, Expressing, Transfection, Pull Down Assay, Western Blot

Journal: Cell Death & Disease

Article Title: LINC01106 post-transcriptionally regulates ELK3 and HOXD8 to promote bladder cancer progression

doi: 10.1038/s41419-020-03236-9

Figure Lengend Snippet: Four groups of J82 and T24 cells were involved, including sh-NC, sh-LINC01116#1, sh-LINC01116#1 + pcDNA3.1/ELK3, sh-LINC01116#1 + pcDNA3.1/ELK3 + pcDNA3.1/HOXD8. a , b Colony formation assay and EdU assay detected the proliferation ability of cells in these four groups. c Flow cytometry analysis detected the apoptosis rate of cells in these four groups. d , e Transwell assays detected the migration and invasion capacity of indicated J28 and T24 cells. f Western blot examined the expression of E-cadherin, N-cadherin, and Vimentin in indicated J28 and T24 cells. * P < 0.05, ** P < 0.01.

Article Snippet: The fixed J82 and T24 cells were rinsed in PBS, dehydrated, and air-dried for cultivating in hybridization buffer with LINC01116-specific RNA fluorescence in situ hybridization (FISH) probe (Ribobio).

Techniques: Colony Assay, EdU Assay, Flow Cytometry, Migration, Western Blot, Expressing

Journal: Cell Death & Disease

Article Title: LINC01106 post-transcriptionally regulates ELK3 and HOXD8 to promote bladder cancer progression

doi: 10.1038/s41419-020-03236-9

Figure Lengend Snippet: a The expression of LINC01116 was examined in J82 and T24 cells transfected with shRNAs targeting HOXD8 or ELK3 by RT-qPCR. b ChIP assay was further conducted to explore the functional relationship between LINC01116 and HOXD8. c Five binding sites between LINC01116 promoter and HOXD8 were predicted using JASPAR software. d Luciferase reporter gene assays were performed to verify which site in LINC01116 promoter was recognized by HOXD8. e Luciferase reporter assays also detected the influence of HOXD8 on the activity of P1. * P < 0.05, ** P < 0.01.

Article Snippet: The fixed J82 and T24 cells were rinsed in PBS, dehydrated, and air-dried for cultivating in hybridization buffer with LINC01116-specific RNA fluorescence in situ hybridization (FISH) probe (Ribobio).

Techniques: Expressing, Transfection, Quantitative RT-PCR, Functional Assay, Binding Assay, Software, Luciferase, Activity Assay

Journal: Cell Death & Disease

Article Title: LINC01106 post-transcriptionally regulates ELK3 and HOXD8 to promote bladder cancer progression

doi: 10.1038/s41419-020-03236-9

Figure Lengend Snippet: The drawing analyzed the two functional pathways of LINC01116-miR-3612-ELK3 and LINC01116-DKC1-HOXD8 in BCa cells. * P < 0.05, ** P < 0.01.

Article Snippet: The fixed J82 and T24 cells were rinsed in PBS, dehydrated, and air-dried for cultivating in hybridization buffer with LINC01116-specific RNA fluorescence in situ hybridization (FISH) probe (Ribobio).

Techniques: Functional Assay

Journal:

Article Title: Analysis of Pathogen-Host Cell Interactions in Purpura Fulminans: Expression of Capsule, Type IV Pili, and PorA by Neisseria meningitidis In Vivo

doi: 10.1128/IAI.70.9.5193-5201.2002

Figure Lengend Snippet: Confocal laser scanning microscopy to detect N. meningitidis in skin biopsy samples from patients with meningococcal disease. Seven-micrometer-thick frozen sections were analyzed as sequential 1-μm-thick sections (×1,000). (A and B) A microcolony of N. meningitidis double stained with anticapsule monoclonal antibody detected with Alexa Fluor 488 (green fluorescence) and antipilin monoclonal antibody detected with Alexa Fluor 594 (red fluorescence), respectively. (C and D) A microcolony staining with antipilin monoclonal antibody (red fluorescence) and some background fluorescence but no specific staining with anticapsule monoclonal antibody (green fluorescence), respectively.

Article Snippet: The sections were then incubated for 1 h at 37°C with the biotin-coupled anticapsule meningococcal antibody (diluted 50 μg/ml in PBS containing 0.2% saponin and 10% FCS), which was then detected by incubation for 1 h at room temperature with a streptavidin-linked fluorochrome (streptavidin-conjugated Alexa Fluor 488 [Molecular Probes], diluted 10 μg/ml in PBS containing 1% FCS).

Techniques: Confocal Laser Scanning Microscopy, Staining, Fluorescence