biological samples human ascites  (Vector Biolabs)

Journal: Biochemistry and Biophysics Reports

Article Title: LIN28-mediated gene regulatory loops synchronize transitions throughout organogenesis

doi: 10.1016/j.bbrep.2025.102226

Figure Lengend Snippet: Transcriptional regulation of LIN28A and LIN28B by developmental transcription factors. A) Schematic of transcription factors that orchestrate germ layer formation and early embryonic lineage specification and differentiation. B) Top: Schematic of morphogen-induced lung/endoderm formation from iPSCs through initiation of lung progenitors in vitro. Bottom: Gene expression analysis of single cell RNA-sequencing of roughly 10,000 cells (close 1200 for each day) throughout morphogen-directed lung endoderm formation from iPSCs. Data from manuscript (Ori et al., 2023). C) Fold change in gene expression of LIN28A and LIN28B upon infection of embryonic lung fibroblast WI-38, with adenoviral constructs: GFP, FOXA2, NKX2-1, SOX2 and SOX9 for 48–72 h before lysing in Trizol. Dots on the graph representative of n = 2 independent experiments in quadruplicate. D) Schematic of development of endoderm differentiation. Transient transfection of txrn factors into HEK293 with construct stably expressing dual luciferase construct; firefly and Renilla luciferase. Validation of a subset of factors that regulated human LIN28A and LIN28B promoter/enhancer sequences during screening experiments in panels B) and C) during E) pluripotency and F) endoderm/lung development. G) progenitor specification. Graphs in E), F) and G) are representative of n = 2 independent validation experiments out of 3. P values for C) were determined using 2-way ANOVA of normal distribution. P values of significance: In the order of left to right for panel C: LIN28A – FOXA2- ∗0.024, NKX2-1- ∗0.035, SOX9- ∗0.021; LIN28B – NKX2-1- ∗0.021, ∗SOX9- 0.021 P values for graphs E), F) and G) were determined using 1-way ANOVA of normal distribution in E) and 1-way ANOVA of lognormal distribution for F) and G). P values of significance: In the order of left to right for each panel: E)∗ 0.0375, ∗∗ 0.0021; ∗ 0.0184, ∗0.0321 F)∗ 0.0434, ∗0.0325; ∗0.0191, ∗0.0477 G)∗∗0.0097,∗ 0.0112; ∗∗0.0021, ∗∗0.0078; ∗∗ 0.0036, ∗∗0.0079; ∗∗0.0072, ∗0.0160. Schematics A), B), D) were made using BioRender.

Article Snippet: Lin28a/b were knocked out post-neurosphere formation via adenoviral infection with Ad-Cre-GFP adenovirus (Vector Biolabs, 1700) or Ad-GFP adenovirus (Vector Biolabs, 1060).

Techniques: In Vitro, Gene Expression, RNA Sequencing, Infection, Construct, Transfection, Stable Transfection, Expressing, Luciferase, Biomarker Discovery