Journal: CNS Neuroscience & Therapeutics

Article Title: Neuron‐specific deletion of presenilin enhancer2 causes progressive astrogliosis and age‐related neurodegeneration in the cortex independent of the Notch signaling

doi: 10.1111/cns.13454

Figure Lengend Snippet: Molecular characterization of forebrain neuron‐specific Pen‐2 cKO mice. A, Co‐staining for NeuN and tdTomato. CaMKIIα‐Cre;Rosa26‐LSL‐tdTomato mice were used to collect brain sections. There was wide expression of tdTomato in the cortex, the hippocampus, the olfactory bulb, and the striatum but not the cerebellum. Most NeuN + cells were positive for tdTomato. The boxed area in (a) was enlarged as (b). Scale bar is 500 μm in (a) but 25 μm in (b). B, Western analysis on Pen‐2. Cortical protein samples for control (Con) and Pen‐2 cKO (cKO) mice at 3 mo were used. There was significant difference on Pen‐2 levels between control and Pen‐2 cKO mice (*** P < 0.001; n = 3‐4 mice per group). C, Relative Pen‐2 mRNA levels. There was significant difference between control (Con) and Pen‐2 cKO (cKO) mice (*** P < 0.001; n = 4 per group). D, Western analyses for APP‐FL, APP‐CTF and nicastrin. Cortical samples at 3 mo were used. There was no significant difference on protein levels of APP‐FL between control (Con) and Pen‐2 cKO (cKO) mice ( P > 0.6; n = 3 per group). There was significant difference on protein levels of nicastrin between control and Pen‐2 cKO mice (*** P < 0.001; n = 3‐4 per group). E, Nissl staining. There was comparable brain size in control and Pen‐2 cKO mice at 2 mo. Scale bar is 1 mm. F‐I, Representative fluorescence IHC images for NeuN (F), MAP2 (G), GFAP (H) and Iba1 (I). Images were taken from cortices of control and Pen‐2 cKO mice at 2 mo. Scale bar is 200 μm

Article Snippet: The following primary antibodies were used: anti‐Pen‐2 (A15172; 1:1000; Abclonal), anti‐nicastrin (34‐9200; 1:500; Invitrogen), anti‐APP (A8717; 1:1000; Sigma‐Aldrich), anti‐Iba1 (382207; 1:1000; ZENBIO), anti‐GFAP (A14673; 1:1000; Abclonal), anti‐GPADH (CW0100; 1:10,000; CWBIO), and anti‐β‐actin (CW0096; 1:10,000, CWBIO).

Techniques: Staining, Expressing, Western Blot, Fluorescence

Journal: CNS Neuroscience & Therapeutics

Article Title: Neuron‐specific deletion of presenilin enhancer2 causes progressive astrogliosis and age‐related neurodegeneration in the cortex independent of the Notch signaling

doi: 10.1111/cns.13454

Figure Lengend Snippet: Age‐related microgliosis in Pen‐2 cKO mice. A, Representative images for fluorescence IHC on Iba1 in the cortex. Mice at 3, 6, and 9 mo were examined. Note that the immuno‐reactivity of Iba1 was not increased in Pen‐2 cKO mice at 3 mo compared with controls. Boxed areas in (a), (b), (e), and (f) were enlarged as (g), (h), (i), and (j), respectively. Scale bar is 200 μm for (a‐f) or 100 μm for (g‐j). B, Averaged number of Iba1 + cells in the cortex. There was significant difference between control (Con) and Pen‐2 cKO (cKO) mice at each age (3 mo: P > 0.2; 6 mo: ** P < 0.01; 9 mo: *** P < 0.001; n = 3‐4 mice per group). C, Western analysis on Iba1. Note that Iba1 levels in Pen‐2 cKO (cKO) mice did not differ from those in controls (Con) at 3 mo of age but were increased at 6 and 9 mo (3 mo: not significant; 6 mo: *, P < 0.05; 9 mo: ** P < 0.01; n = 3‐4 per group). D‐F, qPCR analyses for Iba1 (D), IL‐1β (E), and TNFα (F). Data for 6 and 9 mo were presented. There was significant increase on levels for Iba1 (D), IL‐1β (E), and TNFα (F) in Pen‐2 cKO (cKO) mice compared with controls (Con) at either age (*** P < 0.001; n = 3‐4 per group)

Article Snippet: The following primary antibodies were used: anti‐Pen‐2 (A15172; 1:1000; Abclonal), anti‐nicastrin (34‐9200; 1:500; Invitrogen), anti‐APP (A8717; 1:1000; Sigma‐Aldrich), anti‐Iba1 (382207; 1:1000; ZENBIO), anti‐GFAP (A14673; 1:1000; Abclonal), anti‐GPADH (CW0100; 1:10,000; CWBIO), and anti‐β‐actin (CW0096; 1:10,000, CWBIO).

Techniques: Fluorescence, Western Blot

Journal: CNS Neuroscience & Therapeutics

Article Title: Neuron‐specific deletion of presenilin enhancer2 causes progressive astrogliosis and age‐related neurodegeneration in the cortex independent of the Notch signaling

doi: 10.1111/cns.13454

Figure Lengend Snippet: NICD does not reduce gliosis in Pen‐2 cKO mice. A‐B, Representative images for fluorescence IHC on GFAP (A) and Iba1 (B). Images were taken from the cortex of control, Pen‐2 cKO or Pen‐2 cKO; NICD mice. Boxed areas in (b,c) were enlarged as (d,e). Scale bar is 200 μm in (a‐c) or 100 μm in (d‐e). C, Averaged number of GFAP + cells in the cortex. There was no significant difference between Pen‐2 cKO and Pen‐2 cKO; NICD mice. There was significant difference between Pen‐2 cKO and control mice (ns, not significant; *** P < 0.001; n = 3 mice per group). D, Averaged number of Iba1 + cells in the cortex. There was no significant difference between Pen‐2 cKO and Pen‐2 cKO; NICD mice. There was significant difference between Pen‐2 cKO and control mice (ns, not significant; *** P < 0.001; n = 3‐4 per group). E, Western blotting on GFAP and Iba1. Cortical samples of control, Pen‐2 cKO and Pen‐2 cKO; NICD mice were used. Blots for GFAP or Iba1 were comparable between Pen‐2 cKO and Pen‐2 cKO; NICD mice

Article Snippet: The following primary antibodies were used: anti‐Pen‐2 (A15172; 1:1000; Abclonal), anti‐nicastrin (34‐9200; 1:500; Invitrogen), anti‐APP (A8717; 1:1000; Sigma‐Aldrich), anti‐Iba1 (382207; 1:1000; ZENBIO), anti‐GFAP (A14673; 1:1000; Abclonal), anti‐GPADH (CW0100; 1:10,000; CWBIO), and anti‐β‐actin (CW0096; 1:10,000, CWBIO).

Techniques: Fluorescence, Western Blot

Journal: Acta biochimica et biophysica Sinica

Article Title: Excessive apoptosis and ROS induced by ethionine affect neural cell viability and differentiation.

doi: 10.1093/abbs/gmaa093

Figure Lengend Snippet: Figure 2. Ethionine inhibited NSC differentiation into neurons and astrocytes (A) Compared with the control group, ethionine-treated group has smaller neurospheres. Scale bar: 100 μm. (B) Immunofluorescence staining of nestin for NSCs. (C–F) Immunofluorescence staining of GFAP (C,E) and map2 (D,F) for NSCs, indicating that the NSCs have developed into astrocytes and neurons in the control group, respectively, while the differentiation of NSCs in the ethionine treatment group was reduced significantly, as the average percentage of map2-positive neurons and GFAP-positive astrocytes was significantly lower. Scale bar: 100 μm. ∗∗∗∗P<0.0001 vs control group.

Article Snippet: The primary antibodies used were anti-GFAP antibody (1:100, A14673; ABclonal, Wuhan, China), anti-map2 antibody (1:100, A17409; ABclonal), and anti-nestin antibody (1:80, A11861; ABclonal), and the secondary antibodies used were Alexa Fluor 488-conjugated goat anti-rabbit IgG H&L (A11011; Invitrogen, New York, USA).

Techniques: Control, Staining