Journal: PLoS ONE

Article Title: Glycogen Synthase Kinase 3 (GSK3) Inhibitor, SB-216763, Promotes Pluripotency in Mouse Embryonic Stem Cells

doi: 10.1371/journal.pone.0039329

Figure Lengend Snippet: (A) Phase-contrast images of mESCs treated with LIF (1,000 U/mL), or 10, 15, and 20 µM SB-216763 for more than a month. All doses of SB-216763 maintained mESCs as compact, elliptical or circular colonies characteristic of pluripotent cells. Morphology for mESC colonies maintained with SB-216763 was comparable to those in LIF supplemented medium. Scale bars represent 100 µm. (B) Twenty thousand mESCs from each treatment were seeded in a 6-well chamber at each passage and monitored for the percentage of pluripotent-like colonies two days after each passage. Data are expressed as the mean of five fields analyzed ± SEM. The percentage of pluripotent-like colonies in vehicle-treated cultures was significantly different from 15 µM and 20 µM SB-216763 samples at passage 4 (p<0.05) and from all treated samples from passage 5 onwards (p<0.05). (C) The average number of the total colonies and total pluripotent-like colonies observed between passages 4–7. The number of pluripotent-like colonies in vehicle-treated cultures was significantly lower than that for all other treatments (p<0.05). None of the SB-216763-treated samples were significantly different from LIF-treated mESCs for either the total number of colonies or the total number of pluripotent-like colonies.

Article Snippet: J1 mESC (SCRC-1010, ATCC) and MilliTrace Nanog GFP Reporter mESC (SCR089, Millipore) cell lines were cultured on MEFs that were mitotically inactivated using 10 μg/mL mitomycin C (Sigma).

Techniques:

Journal: PLoS ONE

Article Title: Glycogen Synthase Kinase 3 (GSK3) Inhibitor, SB-216763, Promotes Pluripotency in Mouse Embryonic Stem Cells

doi: 10.1371/journal.pone.0039329

Figure Lengend Snippet: The graph also shows expression of these markers in EBs derived from month-old LIF and SB-216763 mESCs. SYBR Green qPCR results for these four genes were normalized to β-actin housekeeping gene using qbasePLUS software. SB-216763 treated mESCs expressed Lrh-1 and Sox2 about 3-fold higher than LIF treated mESCs. Nanog expression was also moderately higher in the SB-216763-treated mESCs, whereas Oct-4 expression was nearly 50% higher in LIF-treated mESC. Expression of these genes in differentiated cells was generally lower than the mESCs maintained with either LIF or 10 µM SB-216763. The data are average ± S.D. of technical replicates from one of the two experiments.

Article Snippet: J1 mESC (SCRC-1010, ATCC) and MilliTrace Nanog GFP Reporter mESC (SCR089, Millipore) cell lines were cultured on MEFs that were mitotically inactivated using 10 μg/mL mitomycin C (Sigma).

Techniques: Expressing, Derivative Assay, SYBR Green Assay, Software

Journal: PLoS ONE

Article Title: Glycogen Synthase Kinase 3 (GSK3) Inhibitor, SB-216763, Promotes Pluripotency in Mouse Embryonic Stem Cells

doi: 10.1371/journal.pone.0039329

Figure Lengend Snippet: MESCs maintained with either LIF or 10 µM SB-216763 for more than a month yielded pluripotent-like colonies. Immunostaining revealed that Nanog expression (green fluorescence) was homogeneous in mESC colonies maintained with SB-216763, whereas LIF-treated mESC colonies often displayed incomplete Nanog staining. Three representative colonies from each treatment are shown. Scale bars represent 40 µm.

Article Snippet: J1 mESC (SCRC-1010, ATCC) and MilliTrace Nanog GFP Reporter mESC (SCR089, Millipore) cell lines were cultured on MEFs that were mitotically inactivated using 10 μg/mL mitomycin C (Sigma).

Techniques: Immunostaining, Expressing, Fluorescence, Staining

Journal: Cell reports

Article Title: Systematic Discovery of Endogenous Human Ribonucleoprotein Complexes

doi: 10.1016/j.celrep.2019.09.060

Figure Lengend Snippet:

Article Snippet: Mouse J1 ES , ATCC , SCRC-1010.

Techniques: Recombinant, Gene Expression, Software

Journal: Frontiers in Cellular Neuroscience

Article Title: Adipose-derived stem cell-conditioned medium mitigates ischemia-induced neuronal injury via the JAK1/STAT3 signaling pathway

doi: 10.3389/fncel.2026.1744887

Figure Lengend Snippet: ADSC-CM promotes neurite outgrowth in OGD-injured neurons through the JAK1-STAT3 signaling pathway. (A) On day 5 of cultivation, neuronal neurites extended and formed an extensive neural network. Immunofluorescence staining showed that the neurons were positive for the neuronal-specific marker Tuj1. After OGD injury, neuronal neurites were damaged, partially interrupted, and disappeared. (B) Western blot bands of JAK1, pJAK1, STAT3, pSTAT3, and GAPDH for each group. (C) Comparison of the relative ratio of pJAK1 to JAK1 across experimental groups ( n = 4). (D) Comparison of the relative ratio of pSTAT3 to STAT3 across experimental groups ( n = 4). (E) Immunofluorescence images of neurons from each group, showing the effects of ADSC-CM and GLPG0634 on neurite outgrowth. Neurons were stained with the Tuj1 antibody (red) to label the neuronal cytoskeleton, and nuclei were counterstained with DAPI (blue). (F) Diagram illustrating how to measure the length of the longest neurite and the number of primary neurites in neurons. The green line shows the trajectory of the longest neurite, and white arrows point to primary neurites. (G,H) Comparison of the length of the longest neurite and the number of primary neurites in neurons across experimental groups ( n = 4). Data are expressed as means ± SEM. The difference between the groups was assessed using a one-way ANOVA followed by Bonferroni post hoc tests. * p < 0.05, ** p < 0.01, *** p < 0.001. Scale bars: 100 μm (A) , 20 μm (E,F) .

Article Snippet: After overnight incubation with rabbit anti-Tuj1 antibody (1:200, BM3881, Boster, Wuhan, China) at 4 °C, cells were washed and then incubated with Cy3-conjugated goat anti-rabbit IgG (1:400, AS007, ABclonal) for 1 h at room temperature in the dark.

Techniques: Immunofluorescence, Staining, Marker, Western Blot, Comparison