Journal: Cell Death & Disease

Article Title: Rb substantially compensates for the double loss of p130 and p107 in adult but not embryonic neural stem cell lineages

doi: 10.1038/s41419-025-07815-6

Figure Lengend Snippet: IHC on brain sagittal sections in WT ( A – A”’ , E ), RbKO ( B – B”’ , F ), DKO ( C – C”’ , G ), and TKO ( D – D”’ , H ) embryos showing Hes1 and Hes3 staining throughout the DC in the latter two genotypes only. A' – D”’ Higher magnification images of boxed areas shown in ( A – D ). The majority of Hes-positive cells co-express AC-3 (but not Ki67; data not shown). I Quantification of cell counts inside the DC at E14.5. A very low number of or no Hes1+ cells were detected in WT and RbKO mice. Scale bars, 50 µm. Error bars, mean ± SD. Unpaired 2-tailed Student’s t-test; *p < 0.05, **p < 0.01, ***p < 0.001. n = 3 biological replicates.

Article Snippet: The primary antibodies used are: Hes1 (mouse, 1:20, DSHB PCRP-HES1-2E8-s) and Hes3 (mouse, 1:20, DSHB PCRP-HES3-1A10-s), vimentin (mouse, 1:20, DSHB 40E-C-S), and PH3 (rabbit, 1:1000, Millipore 06-570).

Techniques: Staining

Journal: Brain

Article Title: Notch-1 signalling is activated in brain arteriovenous malformations in humans

doi: 10.1093/brain/awp246

Figure Lengend Snippet: Immunohistochemical detection of Notch-1 ligands and Hes-1 in human brain AVMs. (A) Dll4, Jagged-1 and Hes-1 expression (black) was increased in human brain AVMs (left panels, low-magnification; middle panels, high-magnification) compared with normal human middle cerebral artery (right panels). A few non-vascular cells in normal control brain also expressed Hes-1 (arrow). (B) Notch1, Jagged-1, Dll4, NICD and Hes-1 were expressed or activated in vascular cells of E17 mouse brain. (C) Immunostaining was performed on H9 human embryonic stem cells using (left to right) anti-Jagged-1 (green) and anti-NICD (red); anti-Notch-1 (green); anti-Dll4 (green) and anti-Hes1 (green). DAPI (blue) was used to counterstain nuclei.

Article Snippet: Primary antibodies used were (i) goat or rabbit polyclonal anti-Notch-1 (Santa Cruz Biotechnology, 1:100), (ii) rabbit polyclonal anti-NICD (intracellular domain of Notch; Abcam, 1:500), (iii) goat polyclonal anti-Jagged-1 (Santa Cruz Biotechnology, 1:100), (iv) rabbit polyclonal anti-Hes-1 (Chemicon, 1:500), (v) mouse monoclonal anti-Delta-like-4 (Dll4; R&D, 1:200), (vi) mouse monoclonal anti-BrdU (bromodeoxyuridine; Roche, 2 µg/ml), (vii) rabbit polyclonal anti-extracellular domain of Dll4 (Ab7280; Abcam, 1:500) and (vii) rabbit anti-smooth muscle α-actin (αSMC; Maine Biotechnology Service, 1:300).

Techniques: Immunohistochemical staining, Expressing, Immunostaining

Journal: Neural Regeneration Research

Article Title: Protein hairy enhancer of split-1 expression during differentiation of muscle-derived stem cells into neuron-like cells ★

doi: 10.3969/j.issn.1673-5374.2012.028.003

Figure Lengend Snippet: Expression of hairy enhancer of split-1 (Hes1) protein (specific staining of muscle-derived stem cells) after differentiation in the experimental group and the control group (immunofluorescence cytochemistry staining, fluorescence microscopy, × 400). (A) In the control group, the red fluorescence (Hes1) was strong (arrows), and nuclei stained blue. (B) The cytoplasm of neuron-like cells was negative for Hes1. There was no Hes1 present in the experimental group and blue staining was very strong in the nucleus (arrows).

Article Snippet: Cells were then incubated in confining liquid for 20 minutes, followed by incubation with rabbit anti-rat Hes1 polyclonal primary antibody (1:100; Santa Cruz Biotechnology) at 4°C overnight.

Techniques: Expressing, Staining, Derivative Assay, Control, Immunofluorescence, Fluorescence, Microscopy

Journal: Neural Regeneration Research

Article Title: Protein hairy enhancer of split-1 expression during differentiation of muscle-derived stem cells into neuron-like cells ★

doi: 10.3969/j.issn.1673-5374.2012.028.003

Figure Lengend Snippet: Hes1 protein expression as detected by western blot assay during the differentiation of muscle-derived stem cells into neurons. (A) Western blot analysis showed that Hes1 was down-regulated in neuron-like cells induced after 6 days in culture. (B) Western blot semi-quantitative analysis results. The data (absorbance ratio to β-actin) are expressed as mean ± SD; the experiment was repeated five times. a P < 0.05, vs . control group using two samples t -test. 1: Experimental group; 2: control group.

Article Snippet: Cells were then incubated in confining liquid for 20 minutes, followed by incubation with rabbit anti-rat Hes1 polyclonal primary antibody (1:100; Santa Cruz Biotechnology) at 4°C overnight.

Techniques: Expressing, Western Blot, Derivative Assay, Control

Journal: PLOS Genetics

Article Title: Her6 and Prox1a are novel regulators of photoreceptor regeneration in the zebrafish retina

doi: 10.1371/journal.pgen.1011010

Figure Lengend Snippet: (A, B) Schematic representation of retinal cells and observed regenerative processes and timeline. Rod (purple) and cone (blue) photoreceptors (PR) are located in the outer nuclear layer (ONL). Müller glia (MG) are located in the inner nuclear layer (INL) with processes spanning across the retina from the ONL through to the GCL (ganglion cell layer). Following PR ablation, MG are activated and dedifferentiate into progenitors, which proliferate and migrate prior to differentiation into photoreceptors by 96 hours post ablation (hpa). (C-H”) In adult retina, during the 96-hour regenerative time span following the ablation of PRs (red), proliferation is marked by proliferative cell nuclear antigen (PCNA, pink) starts at 24 hpa. Expression of Hairy-enhancer of split (Hes1, green) is seen both in the control and following the ablation in the inner half of the INL and in the GCL. Focusing on the proliferative cells, the expression of Hes1 is downregulated as PCNA expression increases from 24 hpa onwards. DAPI (cyan) marks the retinal layers. Scale bars: 50 μm.(I) UMAP plot of FACS MG 3 days after injury is subdivided into distinct MG derived cell populations based on stereotypical markers. (J) The feature plots show that her6 is expressed mainly in the quiescent and some proliferating MG ( pcna , ascl1a expressing) subsets and downregulated in differentiating ( pcna , insm1a expressing) cell populations. (K) Summary of Hes1/ her6 expression during regeneration. Hes1/ her6 is present in the initial progenitors and downregulated upon their proliferation.

Article Snippet: Primary antibodies used were mouse anti-Prox1a (1:2000–1:10000, Invitrogen), mouse anti-GS (1:500, Merck MAB302), rabbit anti-PCNA (1:400, Sigma SAB2701819), rabbit anti-Hes1 (1:500, Invitrogen #MA5-32258), mouse anti-zpr1 (1:500, Zebrafish International Resource Centre), rabbit Anti-Hes4 (1: 500, Invitrogen #PA5-84551).

Techniques: Expressing, Derivative Assay

Journal: PLOS Genetics

Article Title: Her6 and Prox1a are novel regulators of photoreceptor regeneration in the zebrafish retina

doi: 10.1371/journal.pgen.1011010

Figure Lengend Snippet: (A-D’) Downregulation of hairy-related 6 ( her6 ) by morpholino (MO) in adult retina resulted in a significant increase of Prospero homeobox 1a (Prox1a) in the ONL during early phases of regeneration at 48hpa (SC n = 15, 5.294 ±0.3032, her6 MO n = 12, 9.507±1.339), but not at 72hpa (SC n = 9, 16.97 ±3.096, her6 MO n = 9, 11.34 ±2.625), quantified in E. (F-I’) Knockdown of Prox1a using MO led to a significant downregulation of Hes1/Her6 expression at 120 hpa (SC n = 11, 9.355 ±1.792, prox1a MO n = 6, 3.255 ±0.4484), but not at 72 hpa (SC n = 13, 9.958 ±0.9212, prox1a MO n = 14, 6.929±1.044), quantified in J. (K) Summary of expression and cross-regulation between Her6 and Prox1a expression during regeneration. Her6 is expressed during progenitor formation while Prox1a is present in differentiating PRs. Her6 is a negative regulator of Prox1a and Prox1a is a positive regulator of Her6. Data are represented as mean ± SEM. *p < 0.05. Scale bars: 50 μm.

Article Snippet: Primary antibodies used were mouse anti-Prox1a (1:2000–1:10000, Invitrogen), mouse anti-GS (1:500, Merck MAB302), rabbit anti-PCNA (1:400, Sigma SAB2701819), rabbit anti-Hes1 (1:500, Invitrogen #MA5-32258), mouse anti-zpr1 (1:500, Zebrafish International Resource Centre), rabbit Anti-Hes4 (1: 500, Invitrogen #PA5-84551).

Techniques: Expressing