Journal: Cells

Article Title: Fixed-Bed Bioreactor Culture Enhances Yield and Reparative Properties of hTERT Mesenchymal Stem Cell Extracellular Vesicles

doi: 10.3390/cells15070654

Figure Lengend Snippet: EV function and signaling activity. ( a ) Primary keratinocytes were exposed to IR and treated with EVs from either flask or bioreactor cultures. Cell viability was assessed at 24, 48, and 72 h using the CellTiter-Glo assay. ( b ) Non-irradiated keratinocytes were treated with EVs from flask or bioreactor cultures, and viability was assessed after 72 h using the same assay. * = p < 0.05; *** = p < 0.001; **** = p < 0.0001. ( c ) Western blot of cell cycle proteins associated with hTERT MSC EVs produced from both the bioreactor and flask-based cultures. ( d ) In vitro kinase assay of EV-associated CDK2. EVs were immunoprecipitated with CDK2 antibodies or IgG control and kinase activity was assessed using Histone H1 as substrate.

Article Snippet: Normal primary epidermal keratinocytes (ATCC ® PCS-200-011TM) were maintained in Dermal Cell Basal Medium (ATCC ® PCS-200-030TM) supplemented with Keratinocyte Growth Kit (ATCC ® PCS-200-040TM).

Techniques: Activity Assay, Glo Assay, Irradiation, Western Blot, Produced, In Vitro, Kinase Assay, Immunoprecipitation, Control

Journal: bioRxiv

Article Title: Regulation and Function of the HPV16 CircE7 RNA

doi: 10.64898/2026.03.09.710444

Figure Lengend Snippet: (A) RT-qPCR of circE7 and E6*I normalized to linear E6 RNA from primary keratinocytes transfected with HPV16 WT or HPV16 mut2 genomes (circE7 splicing acceptor m 6 A motif mutation, no change to AA sequence). *p<0.05, ****p<0.0001. (B) Western blot of p53, GAPDH and HSP90 in whole cell extracts from Keratinocytes transfected with HPV16 WT or mut2 genomes. (C) Viral DNA detected by qPCR after HIRT DNA extraction out of primary Keratinocytes transfected with either wild type or mut2 HPV16. ****p<0.0001. (D) Crystal Violet staining of keratinocytes grown in 10% BCS DMEM for 2-6 weeks. Cells were transfected with either wild type HPV16 or HPV16 Mut2. Representative images (left) and quantification at week 6 (right). ***p<0.001

Article Snippet: Briefly, early passage primary keratinocytes were co-transfected with an equal DNA ratio of either pHPV16ANsL WT or Mut2 and pCAGGS-nlsCre plasmids (gift from CM Chiang lab) using TransIT Keratinocyte Transfection Reagent (MirusBio, MIR 2800) according to the manufacturer’s protocol.

Techniques: Quantitative RT-PCR, Transfection, Mutagenesis, Sequencing, Western Blot, DNA Extraction, Staining

Journal: Journal of Biological Chemistry

Article Title: Phosphorylated TP63 Induces Transcription of RPN13, Leading to NOS2 Protein Degradation

doi: 10.1074/jbc.m110.158642

Figure Lengend Snippet: FIGURE 1. Cisplatin induces RPN13 expression at the RNA level. Wild- type Np63 and Np63-S385G cells were treated with control medium () or 10 g/ml cisplatin (CIS; ) for 12 h. GAPDH was used as a loading control. A, RT-PCR analysis for RPN13 transcription. B, qPCR. Values for RPN13 (in relative units (RU)) were normalized to values for GAPDH, and values obtained from the control untreated samples were designated as 1. Experiments were performed in triplicate. FIGURE 2. Schematic representation of the human RPN13 gene pro- moter. The sequence of the human 1500-bp RPN13 promoter was found on the UCSC Genome Bioinformatics human genome web site, and certain potential TF-responsive elements (RE) were defined using TFSEARCH soft- ware. TF sequences are shown in boldface. The TSS is shown as an uppercase letter. The following responsive elements were located in the RPN13 pro- moter: TP63 (1376/1354, 1231/1216, 1189/1167, and 500/ 481), NF-Y/DDIT3 (1267/1246, 93/71, and 65/37), NF-B (995/ 985 and 724/716), STAT (857/843), and GAS (gamma-activated site; 971/862).

Article Snippet: Antibodies—We used a rabbit anti- Np63 polyclonal antibody (Ab-1, EMD Chemicals); a rabbit anti-TP63 (TP73L) monoclonal antibody (clone Y289, NB110-57309) and a rabbit anti-DDIT3 (DNA damage-inducible transcript 3) polyclonal antibody (NB100-78344) (Novus Biologicals); mouse monoclonal antibodies against -actin (Sigma) and NF-YA (Rockland Immunochemicals); and rabbit polyclonal antibodies against DNA topoisomerase II (TOP2A, ab74715), RPN13 (ab91567), NF- B p65 subunit (ab32536), NF- B p50 subunit (ab7549), STAT3 (signal transduction activator of transcription 3; ab32500), and UCH37 (ab38528) (Abcam).

Techniques: Expressing, Control, Reverse Transcription Polymerase Chain Reaction, Sequencing