Journal: International Journal of Molecular Medicine

Article Title: Bmi1 regulates neural differentiation of mesenchymal stem cells through the Wnt3a-RhoA signaling pathway to repair ischemic brain injury in rats

doi: 10.3892/ijmm.2025.5596

Figure Lengend Snippet: Effects of B lymphoma Mo-MLV insertion region 1 homolog and Wnt3a on the proliferation of MSCs. (A) Morphology of primary MSCs inoculated into culture flasks. Morphology of MSCs in primary culture for (B) 3 and (C) 7 days. (D) Morphological characterization of third-generation MSCs. Effects of different concentrations of (E) Wnt3a and (F) PTC209 on the proliferation of MSCs. ** P<0.01 vs. 0 μ M group. MSCs, mesenchymal stem cells.

Article Snippet: Wnt3a recombinant cytokine (cat. no. 96-315-20-10) was purchased from PeproTech; Thermo Fisher Scientific, Inc. and the small molecule inhibitor PTC209 (cat. n. S7372) was purchased from Selleck Chemicals.

Techniques:

Journal: International Journal of Molecular Medicine

Article Title: Bmi1 regulates neural differentiation of mesenchymal stem cells through the Wnt3a-RhoA signaling pathway to repair ischemic brain injury in rats

doi: 10.3892/ijmm.2025.5596

Figure Lengend Snippet: The mechanism underlying the effects of Bmi1 on MSC transplantation-induced ischemic brain injury repair. In vitro experiments used Wnt3a and PTC209 to treat MSCs to explore the roles and regulatory mechanisms of the Bmi1 and Wnt3a-Rhoa signaling pathways in the neural differentiation of MSCs. In vivo experiments were conducted to investigate the role of the Bmi1 and Wnt3a-RhoA signaling pathway in brain injury repair in MCAO rats and its mechanism. Different MSCs were transplanted into rat brain tissues after in vitro labeling, after which, the ischemic brain injury in each group was compared and the recovery of neurological function was assessed. Bmi1, B lymphoma Mo-MLV insertion region 1 homolog; H&E, hematoxylin and eosin; IF, immunofluorescence; MSCs, mesenchymal stem cells; NSS, Neurological Severity Score; WB, western blotting.

Article Snippet: Wnt3a recombinant cytokine (cat. no. 96-315-20-10) was purchased from PeproTech; Thermo Fisher Scientific, Inc. and the small molecule inhibitor PTC209 (cat. n. S7372) was purchased from Selleck Chemicals.

Techniques: Transplantation Assay, In Vitro, Protein-Protein interactions, In Vivo, Labeling, Immunofluorescence, Western Blot

Journal: Acta Naturae

Article Title: A Highly Productive CHO Cell Line Secreting Human Blood Clotting Factor IX

doi:

Figure Lengend Snippet: Analysis of target gene integrity in the genomic DNA and cDNA from producer cell lines by PCR, RT-PCR, Southern blotting and pseudo-Northern blotting. DNA ladder bands are shown as bps. Panel A – amplification of whole ORF regions of the target genes in genomic DNA (gDNA) by PCR. The annealing temperature gradient is shown with red triangles; the linear shift in annealing temperature was from 53 to 68 oC. The expected amplification products of the FIX , furin, and VKORC1 genes are shown with red arrows. Panel B – RT-PCR products for total mRNA with primers towards the 5’- and 3’- ends of the FIX ORF; the amplification product of expected size is shown with a red arrow. Panel C – Southern blotting of gDNA with biotin-labeled probes. The IDbla probe is the probe targeting the IRES and DHFR regions, the ampicillin resistance gene (the bla gene) and the region of plasmid origin of replication (ori). The FIX probe is the probe targeting the FIX ORF region. Sizes of the detected target restriction fragments are shown with arrows. Negative agarose gel images; the membrane images were contrast-enhanced to add visibility. Panel D – Analysis of cDNA by pseudo-Northern blotting. Denotation is the same as that in panel C. The actual position and size of FIX cDNA are shown with an arrow

Article Snippet: The commercially available clone of human FIX cDNA, pCMV6-XL4/NM_000133.2 (sc126517, Origene, USA), and adapter primers AD-9-AbsF and AD-9- NheR ( ) were used as a source of FIX ORF.

Techniques: Reverse Transcription Polymerase Chain Reaction, Southern Blot, Northern Blot, Amplification, Labeling, Plasmid Preparation, Agarose Gel Electrophoresis, Membrane