Journal: Biology

Article Title: Stage-Specific Alternative Polyadenylation During Human Neural Differentiation Revealed by Integrated Long- and Short-Read Sequencing

doi: 10.3390/biology15010024

Figure Lengend Snippet: Immunofluorescence staining showing the expression of neural stem/progenitor cell markers in H9-derived cells. The cell nuclei are labeled with DAPI (blue). Nestin was detected in green. OCT4, NANOG, and SOX1 were detected in red. PAX6 was detected in red in NSCs and in green in NPCs. Scale bar = 50 μm. Original figures see .

Article Snippet: Cells were fixed with 4% paraformaldehyde (PFA) at room temperature for 20 min, permeabilized with 0.25% Triton X-100 for 30 min and blocked with 5% donkey serum for 1 hr before incubating with primary antibodies including OCT4 (1:200, 962649, R&D System, Minneapolis, MN, USA), NANOG (1:100, 14295-1-AP, Proteintech, Rosemont, IL, USA), PAX6 (1:200, PRB-278P, BioLegend, San Diego, CA, USA), NESTIN (1:500, 809801, BioLegend), and SOX1 (1:200, 67994-1-Ig, Proteintech) at 4 °C overnight.

Techniques: Immunofluorescence, Staining, Expressing, Derivative Assay, Labeling

Journal: JCI Insight

Article Title: The PERK/ATF4 pathway is required for metabolic reprogramming and progressive lung fibrosis

doi: 10.1172/jci.insight.189330

Figure Lengend Snippet: ( A ) Lung macrophages from normal and asbestosis humans were obtained by bronchoalveolar lavage (BAL) and subjected to immunoblot analysis ( n = 4). Densitometry of ( B ) phosphorylated (p-) PERK, ( C ) p-eIF2α, and ( D ) p-IRE1α in humans. ( E ) WT mice were exposed to man-made vitreous fiber (MMVF) or asbestos (100 μg intratracheally; i.t.). BAL was performed on day 21, and lung macrophages were subjected to immunoblot analysis ( n = 3). Densitometry of ( F ) p-PERK, ( G ) p-eIF2α, and ( H ) p-IRE1α from exposed mice. ( I ) Macrophages were exposed to vehicle (Con), asbestos (Asb), tunicamycin (TUN), thapsigargin (TH), or 4PBA (PBA) and subjected to immunoblot analysis. Densitometry of ( J ) p-PERK and ( K ) p-eIF2α ( n = 3). ( L ) Macrophages were exposed to vehicle or asbestos and stained for p-PERK. The staining was imaged by confocal microscopy, scale bars at 10 μm and 40×. ( M ) Quantification of mean fluorescence intensity ( n = 3). Data shown as mean ± SEM. Two-tailed Student’s t test in B – D , F – H , and M . One-way ANOVA with Tukey’s post hoc comparison in J and K . * P ≤ 0.05, ** P ≤ 0.01, and **** P ≤ 0.0001.

Article Snippet: The pcDNA3.1 (Invitrogen), PERK WT (plasmid 21814), PERK DN (plasmid 36954), and IRE1α WT (plasmid 20744) vectors were purchased from Addgene.

Techniques: Western Blot, Staining, Confocal Microscopy, Fluorescence, Two Tailed Test, Comparison