Journal: Science advances

Article Title: Significance of birth in the maintenance of quiescent neural stem cells.

doi: 10.1126/sciadv.adn6377

Figure Lengend Snippet: Fig. 2. Preterm birth RG transiently adopt a neurogenic state. (A) Experimental scheme. (B) Representative images of coronal V-SVZ sections stained for Glul RNA (green) at E18.5, full-term P2, and preterm P3. Nuclei were stained with DAPI (blue). (C) Percentages of Glul RNA dots area in the V-SVZ area (n = 3 mice each; one-way ANOVA followed by Tukey-Kramer test). (D) Representative images of coronal V-SVZ sections in GFAP-EGFP mice stained for EGFP (green), Glul (red), and Nestin (white) at E18.5, full-term P2, and preterm P3. Yellow lines, traced outlines of RG based on EGFP staining. (E) Relative expression levels of Glul in RG (each n = 60 cells from three mice; Kruskal-Wallis test followed by Steel-Dwass test). (F) Experimental scheme. (G) Representative projection images of coronal V-SVZ sections stained for pS6 (green), Mash1 (red), and Nestin (white) at 18.5, 21.5, and 26.5 dpc. White arrows, Mash1+pS6−Nestin+RG. Yellow arrows, Mash1−pS6+Nestin+ RG. Blue arrowheads, Mash1+pS6+Nestin+ RG. (H) Densities of Mash1+Nestin+ RG in the V-SVZ at 18.5, 21.5, and 26.5 dpc (n = 4 mice each; unpaired t test). (I) Densities of pS6+Nestin+ RG in the V-SVZ at 18.5, 21.5, and 26.5 dpc (n = 3 mice each; unpaired t test). (J) Percentages of Mash1+ cells in pS6+Nestin+ RG and pS6−Nestin+ RG at 21.5 dpc (n = 4 mice each; Mann-Whitney U test). (K) Density of pS6+Mash1+Nestin+ RG in the V-SVZ at 21.5 dpc (n = 4 mice each; unpaired t test). Dpc, days post coitus; Mife, mifepristone; LV, lateral ventricle. *P < 0.05, **P < 0.01, and ***P < 0.001. Scale bars, 20 μm [(B), (D), and (G) (single-channel images)] and 50 μm [(G) (merged images)]. Error bars, mean ± SEM. See also fig. S10.

Article Snippet: The following primary antibodies were used: rabbit anti- GS (Glul) (1:500; Abcam), chicken anti- Nestin (1:1000; Aves Labs, Tigard, OR, USA), rabbit anti- Mash1 (1:1000; Abcam), mouse anti- Mash1 (1:1000; Santa Cruz Biotechnology, Dallas, TX, USA), rabbit anti–phosphoS6 ribosomal protein (pS6) (1:500; Cell Signaling Technology, Beverly, MA, USA), rabbit anti–phospho- p70 S6 kinase (Thr421) (1:500; Thermo Fisher Scientific), guinea pig anti- Dcx (1:500; Millipore), mouse anti- NeuN (1:100; Millipore), rat anti- BrdU (1:100; Abcam), sheep anti- BrdU (1:1000; Fitzgerald, Acton, MA, USA), rat antiGFP (1:500; Nacalai Tesque, Kyoto, Japan), rabbit anti- EGFR (1005) (1:100; Santa Cruz Biotechnology), rabbit anti- DsRed (1:1200; Takara Bio, Kusatsu, Japan), and chicken anti- GFAP (1:1000; Abcam).

Techniques: Staining, Expressing, MANN-WHITNEY

Journal: PLoS ONE

Article Title: Efficient Derivation of Multipotent Neural Stem/Progenitor Cells from Non-Human Primate Embryonic Stem Cells

doi: 10.1371/journal.pone.0049469

Figure Lengend Snippet: ( A ) Marmoset ESC-derived primary and secondary neurospheres were dissociated and allowed to differentiate for 10 days, followed by immunocytochemical analysis of βIII-tubulin (neurons), GFAP (astrocytes), CNPase (oligodendrocytes) and Nestin (undifferentiated neural cells). Scale bars, 50 µm. ( B ) The proportions of cells positive for each cell type-specific marker are presented as the percentage of total cells counted by Hoechst 33258-stained nuclei. Data are presented as the means ± SEM ( n = 3).

Article Snippet: Then, samples were incubated at 4°C overnight with the following antibodies: anti-βIII tubulin (mouse IgG 2b , 1∶500; Sigma), Alexa488-conjugated anti-βIII tubulin (mouse IgG, 1∶2,000, Covance, Princeton NJ), anti-glial fibrillary acidic protein (GFAP) (rabbit IgG, 1∶10,000, Dako, Glostrup, Denmark), anti-2′, 3′-cyclic nucleotide 3′-phosphodiesterase (CNPase) (mouse IgG 1 , 1∶4,000, Sigma), anti-human Nestin (rabbit IgG, 1∶4,000) , anti-myelin basic protein (MBP) (rat IgG, 1∶500, AbD Serotec, Raleigh, NC, USA), anti-O4 (mouse IgM 1∶1,000, Millipore), and anti-platelet-derived growth factor receptor (PDGFR)α (rabbit IgG, 1∶2,000, Santa Cruz, Santa Cruz, CA, USA).

Techniques: Derivative Assay, Marker, Staining

Journal: PLoS ONE

Article Title: Efficient Derivation of Multipotent Neural Stem/Progenitor Cells from Non-Human Primate Embryonic Stem Cells

doi: 10.1371/journal.pone.0049469

Figure Lengend Snippet: NS/PCs derived from EBs treated with both RA and purmorphamine, were transplanted into the right striatum of 6–8-week-old female NOD/SCID mice. 4 weeks after the injection, mice were sacrificed and processed for immunohistochemical analysis with the antibodies against markers for neurons (Hu), astrocytes (GFAP), oligodendrocytes (APC), and neural progenitors (Nestin). The marmoset ESC-derived NS/PCs could give rise to neurons, astrocytes and oligodendrocytes in vivo , as well. Scale bars, 20 µm.

Article Snippet: Then, samples were incubated at 4°C overnight with the following antibodies: anti-βIII tubulin (mouse IgG 2b , 1∶500; Sigma), Alexa488-conjugated anti-βIII tubulin (mouse IgG, 1∶2,000, Covance, Princeton NJ), anti-glial fibrillary acidic protein (GFAP) (rabbit IgG, 1∶10,000, Dako, Glostrup, Denmark), anti-2′, 3′-cyclic nucleotide 3′-phosphodiesterase (CNPase) (mouse IgG 1 , 1∶4,000, Sigma), anti-human Nestin (rabbit IgG, 1∶4,000) , anti-myelin basic protein (MBP) (rat IgG, 1∶500, AbD Serotec, Raleigh, NC, USA), anti-O4 (mouse IgM 1∶1,000, Millipore), and anti-platelet-derived growth factor receptor (PDGFR)α (rabbit IgG, 1∶2,000, Santa Cruz, Santa Cruz, CA, USA).

Techniques: Derivative Assay, Injection, Immunohistochemical staining, In Vivo

Journal: International Journal of Molecular Medicine

Article Title: Pimozide augments bromocriptine lethality in prolactinoma cells and in a xenograft model via the STAT5/cyclin D1 and STAT5/Bcl-xL signaling pathways

doi: 10.3892/ijmm.2020.4784

Figure Lengend Snippet: (A) Representative images of the tumorsphere formation assay of CD133 + nestin + stem-like MMQ cells. Scale bar represents 50 µ m. (B) Immunofluorescence analysis of CD133, nestin or Sox2 (red) and DAPI (blue) in CD133 + nestin + stem-like MMQ cells (magnification, ×10). Scale bar represents 50 µ m. (C) The representative cytometric dot-plots of nestin, CD133 and Sox2 expression in CD133 + nestin + stem-like MMQ cells. (D) Representative western blots of p-STAT5, STAT5, CD133, nestin and Ki67 protein expression in CD133 + nestin + stem-like MMQ cells treated with vehicle control, 10 µ M pimozide, 20 µ M bromocriptine or both drugs combined. Pimozide in combination with bromocriptine suppressed the expression of the tumor stem cell marker proteins CD133 and nestin, compared with either drug alone. Results are presented as the mean ± standard error of the mean. (E) Quantitative results of the CD133 + nestin + stem-like MMQ cell tumorsphere formation assay treated with vehicle control, 10 µ M Pimozide, 20 µ M bromocriptine or both drugs combined for 7 days. The combination treatment suppressed the formation of tumorspheres and growth of the CD133 + nestin + stem-like MMQ cells, compared with bromocriptine alone. Results are presented as the mean ± standard error of the mean. * P<0.05. (F) Quantification of p-STAT5, STAT5, CD133, nestin and Ki67 protein levels in CD133 + nestin + stem-like MMQ cells treated with vehicle control, 10 µ M pimozide, 20 µ M bromocriptine or both drugs combined. Data are presented as the mean ± SD (Student's t-test; * P<0.05). p-, phospho-; CON, control; BRO, bromocriptine; PIM, pimozide; STAT5, signal transducer and activator of transcription 5A; Sox2, SRY-box transcription factor 2; ns, not significant.

Article Snippet: As described in our previous study , MMQ cells were isolated using CD133 (cat. no. 130097049; Miltenyi Biotec, Inc.) and Anti-Nestin Magnetic Beads (cat. no. MB106970-T46; Sino Biological, Inc.).

Techniques: Tube Formation Assay, Immunofluorescence, Expressing, Western Blot, Marker

Journal: Frontiers in Cell and Developmental Biology

Article Title: Establishing and characterizing human stem cells from the apical papilla immortalized by hTERT gene transfer

doi: 10.3389/fcell.2023.1158936

Figure Lengend Snippet: Primary antibodies for Western blotting and IF.

Article Snippet: NESTIN , Rabbit , Bioss , bs-0008R.

Techniques: Western Blot