Journal: bioRxiv

Article Title: Hypoxia causes pancreatic β-cell dysfunction by activating a transcriptional repressor BHLHE40

doi: 10.1101/2022.06.28.498031

Figure Lengend Snippet: ( A ) MA plot of RNA-seq data in Ctrl and B40 OE MIN6 cells. Differentially expressed genes (DEGs; adjusted p value < 0.01) are shown in blue, and others in gray. DEGs functioning as β-cell transcription factors are shown in red (n = 3). ( B ) Gene ontology analysis of RNA-seq data. Downregulated DEGs were used as input. ( C and D ) Expression of DEGs shown in A was confirmed by qRT-PCR in Ctrl and B40 OE MIN6 cells (n = 3; C ) or Ctrl and B40 KD MIN6 cells cultured under 20% or 5% O 2 for 24 hours (n = 3; D ). ( E - F ) Western blot of BHLHE40 and MAFA expression in Ctrl and B40 OE MIN6 cells ( E ) or Ctrl and B40 KD MIN6 cells cultured under 20% or 5% O 2 for 24 hours ( F ). ( G ) qRT-PCR of MAFA target genes in Ctrl and B40 KD MIN6 cells cultured under 20% or 5% O 2 for 24 hours (n = 3). ( H ) Glucose-stimulated insulin secretion in MIN6 cells infected with AAV-green fluorescent protein (GFP) (Ctrl) or AAV- Mafa and cultured under 20% or 5% O 2 for 24 hours (n = 4). ( I ) KCl-stimulated insulin secretion in MIN6 cells infected with AAV-GFP (Ctrl) and AAV- Mafa and cultured under 20% or 5% O 2 for 24 hours (n = 4). Data are mean ± SEM; *p < 0.05, *p < 0.01 and ***p < 0.001 by unpaired two-tailed Student’s t test. β-actin was used as a loading control. Ctrl, control.

Article Snippet: Three days after transfection, AAV-GFP and AAV- Mafa were purified with the AAV pro Extraction Solution Kit (6235, Takara, Shiga, Japan), and the titers were determined with a Quick Titer AAV Quantification Kit (VPK-145, Cell Biolabs, Inc.) according to the manufacturer’s instructions.

Techniques: RNA Sequencing Assay, Expressing, Quantitative RT-PCR, Cell Culture, Western Blot, Infection, Two Tailed Test

Journal: Cell

Article Title: Engineered virus-like particles for efficient in vivo delivery of therapeutic proteins

doi: 10.1016/j.cell.2021.12.021

Figure Lengend Snippet:

Article Snippet: Serum was separated by centrifugation at 2000 g for 15 min and stored at -80 o C. Pcsk9 levels were determined by ELISA using the Mouse Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit (R&D Systems; MPC900) following the manufacturer’s instructions.

Techniques: Virus, Recombinant, Transfection, SYBR Green Assay, DNA Extraction, Multiplex Assay, Purification, Gel Extraction, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Cell Isolation, Titration, Amplification, Sequencing, Software