Journal: Cancer cell

Article Title: Liver Cancer Initiation Requires p53 Inhibition by CD44-Enhanced Growth Factor Signaling

doi: 10.1016/j.ccell.2018.05.003

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Antibody details are provided in the . table ft1 table-wrap mode="anchored" t5 REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies anti-CD44pan (DF1485) Santa Cruz Biotechnology Cat# SC-7297; RRID: AB_627065 anti-CD44pan (IM7) ABDSerotec (Biorad) Cat# MCA4703; RRID: AB_2076194 anti-CD44v6 ABD Serotec (Biorad) Cat# MCA1967; RRID: AB_323213 anti-AFP R&D Systems Cat# AF5369; RRID: AB_2258018 anti-Ki67 Genetex Cat# GTX16667; RRID: AB_422351 anti-F4/80 (Clone: A3-1) Caltag/Thermo Fisher Cat# MA1-91124; RRID: AB_2277854 anti-Clvd.

Techniques: Recombinant, Blocking Assay, In Situ, TUNEL Assay, Polymer, Plasmid Preparation, Amplification, Staining, Extraction, Labeling, In Situ Hybridization, RNAscope, HD Assay, cDNA Synthesis, SYBR Green Assay, Negative Control, Software

Journal: Genes

Article Title: Enhancing Transcriptional Reprogramming of Mesenchymal Glioblastoma with Grainyhead-like 2 and HDAC Inhibitors Leads to Apoptosis and Cell-Cycle Dysregulation

doi: 10.3390/genes14091787

Figure Lengend Snippet: Antibodies.

Article Snippet: The cells were incubated overnight with 10 μg/mL anti-CD44 antibody (R&D systems, BBA10) diluted in 1% BSA in PBS at 4 °C.

Techniques:

Journal: Genes

Article Title: Enhancing Transcriptional Reprogramming of Mesenchymal Glioblastoma with Grainyhead-like 2 and HDAC Inhibitors Leads to Apoptosis and Cell-Cycle Dysregulation

doi: 10.3390/genes14091787

Figure Lengend Snippet: GRHL2 represses mesenchymal proteins in GBM cells. Western blots for GRHL2, Slug, CD44, MMP2, and ZEB1 in LN229 cells with or without doxycycline (200 ng/mL) or 2.5 µM vorinostat (VOR). β-Actin was used as a loading control for protein expression. Graphs depict significant changes in protein expression levels from A. Graphs depict means +/− SEM for n = 3. * p < 0.05 t -test; ** p < 0.01 t -test.

Article Snippet: The cells were incubated overnight with 10 μg/mL anti-CD44 antibody (R&D systems, BBA10) diluted in 1% BSA in PBS at 4 °C.

Techniques: Western Blot, Control, Expressing

Journal: Genes

Article Title: Enhancing Transcriptional Reprogramming of Mesenchymal Glioblastoma with Grainyhead-like 2 and HDAC Inhibitors Leads to Apoptosis and Cell-Cycle Dysregulation

doi: 10.3390/genes14091787

Figure Lengend Snippet: Exogenous GRHL2 expression alters the GBM cell cycle. ( A ) Inducible LN229 cells were treated with dox for 72 h with or without HDACi for the last 24 h and were immunoblotted for indicators of cell-cycle phases pCDC2, pRB, cyclin B1, and p21. GAPDH was used as a loading control for protein expression. ( B ) Quantification of data in A. ( C ) Cell-cycle (propidium iodide) flow cytometry of parental or inducible LN229 treated with dox for 72 h. Graph displays mean values for G1, S, and G2/M phase. ( D ) Cell-cycle (propidium iodide) flow cytometry of iLN229 treated with dox for 3 or 14 days. Arrow indicates > 4 N cells. ( E ) Control or dox-treated (21 days) iLN229 cells were stained with CD44 to visualize the cell membrane and DAPI to visualize nuclei. Cells were then counted for multinucleated cells. White arrowheads identify multinucleated cells. Mean values of multinucleated cells are per 300 cells counted. Graphs depict means +/− SEM for n = 3. * p < 0.05 t -test; ** p < 0.01 t -test; *** p < 0.001 t -test.

Article Snippet: The cells were incubated overnight with 10 μg/mL anti-CD44 antibody (R&D systems, BBA10) diluted in 1% BSA in PBS at 4 °C.

Techniques: Expressing, Control, Flow Cytometry, Staining, Membrane

Journal: Stem Cell Research & Therapy

Article Title: Hyaluronan supports the limbal stem cell phenotype during ex vivo culture

doi: 10.1186/s13287-022-03084-8

Figure Lengend Snippet: Effects of HA on TKE2 cells cultured on differently coated dishes or in the presence of HA as a supplement to the media. TKE2 cells were cultured on differently coated dishes ( A ) or on uncoated dishes with HA, HA + anti-CD44 or 4MU + Hyalase supplemented to the media ( B ). The number of ∆Np63 positive cells and the total number of cells (DAPI) were quantified using QuPath and the percentage of ∆Np63 + cells calculated ( C ). *Represents p ≤ 0.05, scale bar represents 50 μm

Article Snippet: In order to verify the mechanism by which HA supports LESCs, anti-CD44 neutralizing antibody (R&D systems, Minneapolis, USA#AF6127) was added to the culture media in order to block the binding of HA to CD44.

Techniques: Cell Culture

Journal: Stem Cell Research & Therapy

Article Title: Hyaluronan supports the limbal stem cell phenotype during ex vivo culture

doi: 10.1186/s13287-022-03084-8

Figure Lengend Snippet: Effects of HA on the differentiation of TKE2 cells cultured on differently coated dishes or in the presence of HA as a supplement to the media. TKE2 cells were cultured on differently coated dishes ( A ) or on uncoated dishes with HA of different molecular weights, HA + anti-CD44 or 4MU + Hyalase supplemented to the media ( B ) under differentiating conditions. The number of ∆Np63 positive cells and the total number of cells (DAPI) were quantified using Qupath and the percentage of ∆Np63 + cells calculated for TKE cells maintained on differently coated dishes ( C ) and with HA, HA + anti-CD44 and 4MU + Hyalase supplemented to the media ( D ). *Represents p ≤ 0.05, scale bar represents 50 μm

Article Snippet: In order to verify the mechanism by which HA supports LESCs, anti-CD44 neutralizing antibody (R&D systems, Minneapolis, USA#AF6127) was added to the culture media in order to block the binding of HA to CD44.

Techniques: Cell Culture

Journal: International Journal of Biological Sciences

Article Title: MicroRNA-143 Inhibits Migration and Invasion of Human Non-Small-Cell Lung Cancer and Its Relative Mechanism

doi: 10.7150/ijbs.6623

Figure Lengend Snippet: Sequences of oligonucleotides used in the paper.

Article Snippet: Membranes were incubated with blocking buffer for 60 min at room temperature and were then incubated with an Anti-CD44v3 (R&D) antibody (diluted with 500-fold) or Anti-β-actin (Santa Cruz, CA, USA) antibody (diluted with 1000-fold) with Blotto overnight at 4℃.

Techniques:

Journal: International Journal of Biological Sciences

Article Title: MicroRNA-143 Inhibits Migration and Invasion of Human Non-Small-Cell Lung Cancer and Its Relative Mechanism

doi: 10.7150/ijbs.6623

Figure Lengend Snippet: CD44v3 is a novel target of miRNA-143 . (A) The putative target genes and mutant complementary sequences of the CD44v3 mRNA 3'-UTR are shown with the miR-143 sequence. (B) The luciferase activities of five putative target gene reporter vectors cotransfected with P-miR-143 or Pc3.1 were measured by dual luciferase assay ( *P < 0.05). (C) The luciferase activities of the wild type CD44v3 3'-UTR (Wt) and mutant type CD44v3 3'-UTR (Mut) cotransfected with P-miR-143 or Pc3.1 were measured by dual luciferase assay (* P < 0.05). (D) The expression level of CD44v3 mRNA in A549 transfected with P-miR-143 or Pc3.1 was determined by the qRT-PCR assay (* P < 0.05). (E) The expression level of CD44v3 protein in A549 transfected with P-miR-143 or Pc3.1 was determined by western blot assay.

Article Snippet: Membranes were incubated with blocking buffer for 60 min at room temperature and were then incubated with an Anti-CD44v3 (R&D) antibody (diluted with 500-fold) or Anti-β-actin (Santa Cruz, CA, USA) antibody (diluted with 1000-fold) with Blotto overnight at 4℃.

Techniques: Mutagenesis, Sequencing, Luciferase, Expressing, Transfection, Quantitative RT-PCR, Western Blot

Journal: International Journal of Biological Sciences

Article Title: MicroRNA-143 Inhibits Migration and Invasion of Human Non-Small-Cell Lung Cancer and Its Relative Mechanism

doi: 10.7150/ijbs.6623

Figure Lengend Snippet: miR-143 overexpression and CD44v3 knockdown show similar phenotypes in suppressing migration and invasion of NSCLC cells. (A) The expression levels of CD44v3 in A549, 95D, H460 and H1299 were determined by the qRT-PCR assay. (B) The CD44v3 mRNA level in A549 transfected with P-miR-143 or negative control (NC) was determined by the qRT-PCR assay. (C) The CD44v3 protein level in A549 transfected with P-miR-143 or negative control (NC) was determined by western blot assay. (D) Transwell assay with matrigel was performed to detect the invasion activity of A549 cells transfected with P-miR-143, Si-CD44v3 and negative control (NC). (E) Transwell assay without matrigel was used to detect the migration activity of A549 cells after transfection (*P < 0.05).

Article Snippet: Membranes were incubated with blocking buffer for 60 min at room temperature and were then incubated with an Anti-CD44v3 (R&D) antibody (diluted with 500-fold) or Anti-β-actin (Santa Cruz, CA, USA) antibody (diluted with 1000-fold) with Blotto overnight at 4℃.

Techniques: Over Expression, Migration, Expressing, Quantitative RT-PCR, Transfection, Negative Control, Western Blot, Transwell Assay, Activity Assay