Journal: eLife

Article Title: A single-cell atlas of the cycling murine ovary

doi: 10.7554/elife.77239

Figure Lengend Snippet: Figure 6. Identification and validation of new secreted estrous staging markers. (A) Expression of granulosa cell transcripts varying by estrous cycle stage. (B) Validation of significantly up- and downregulated transcripts of secreted estrous staging markers by qPCR (n=5 per group, mean ± SEM, *p<0.05, **p<0.01, and ***p<0.005). (C) Localization of estrous staging markers by in situ hybridization (RNAscope) in ovarian sections. (D) Quantification of circulating estrous staging markers proteins in the blood by enzyme-Linked Immunosorbent assay (ELISA) (n=5 per group, mean ± SEM, *p<0.05, and ***p<0.005). (E) Summary of the timing of expression of estrous staging markers in the blood.

Article Snippet: Reagent type (species) or resource Designation Source or reference Identifiers Additional information Genetic reagent (Mus musculus) C57BL/6- Tg(UBC- GFP)30Scha/J Jackson Laboratory stock #004353 Antibody Smooth muscle alpha action (SMA) (Rabbit polyclonal) Abcam #5694 Dilution: 1:300 Commercial assay or kit ACTIVIN A commercial ELISA RnD systems #DAC00B Commercial assay or kit NPPC commercial ELISA Novus Bio #NBP2- 75790 Commercial assay or kit Tinagl1 commercial ELISA LS- Bio #LS- F49684 Commercial assay or kit PRSS35 commercial ELISA Mybiosource #MBS9717242 Commercial assay or kit RNA scope 2.5 HD Duplex detection kit ACD bio #322500 Commercial assay or kit RNA scope 2.5 HD red detection kit ACD bio #322360 Commercial assay or kit The target retrieval and protease plus reagents ACD bio #322330 Other Cdkn1a (M. musculus) NM_007669.4 ACD bio # 408551 RNAscope probe Other Cxcl14 (M. musculus) NM_019568.2 ACD bio #459741 RNAscope probe Morris, Meinsohn et al. eLife 2022;11:e77239.

Techniques: Biomarker Discovery, Expressing, In Situ Hybridization, RNAscope, Enzyme-linked Immunosorbent Assay

Journal: Acta neuropathologica

Article Title: The active contribution of OPCs to neuroinflammation is mediated by LRP1

doi: 10.1007/s00401-019-02073-1

Figure Lengend Snippet: (a) Staining for PDGFRα and LRP1 in the corpus callosum in Olig1cre and Lrp1fl/flOlig1cre mice (arrows = LRP1+ PDGFRα- cells). Dashed line demarcates the border between the cortex and corpus callosum. Scale bar 25μm. (b) Representative immunoblot and (c,d) densitometry analysis for CNP, MBP, and actin from cerebrum and cerebellum (unpaired t-test, MBP *p=0.0329, CNP *p=0.0150; n=5 mice per genotype, 2 independent experiments combined; error bars represent +/− SEM). (e) Representative TEM images and (f) calculated g-ratio of optic nerves (6,000x magnification; n=4 mice per genotype, 10 fields per mouse; linear regression analysis with slopes comparison).

Article Snippet: Primary antibodies used for immunoblotting were against LRP1 (L2170, 1:1000, Sigma), MBP (SMI-94, 1:1000, Covance), CNP (CNP, 1:2000, Aves Labs), actin (A2228, 1:5000, Sigma).

Techniques: Staining, Western Blot, Comparison

Journal: Acta neuropathologica

Article Title: The active contribution of OPCs to neuroinflammation is mediated by LRP1

doi: 10.1007/s00401-019-02073-1

Figure Lengend Snippet: (a) Representative images and (b) quantification of MBP expression in the remyelinating corpus callosum of Olig1cre and Lrp1fl/flOlig1cre mice after 0.5 Wk Rem (unpaired t-test; 2 independent experiments combined; scale bar 100μm). (c) Representative TEM images and (d) quantification of axons in the remyelinating corpus callosum after 0.5 Wk Rem (unpaired t-test; n=4–5 mice per genotype; scale bar 2μm). (e-g) qPCR for Mrf, Mbp, and Cnp from primary OPCs cultured in proliferation (OPC) or differentiation (OLG) media (unpaired t-test; n=3 mice per genotype). (h) Quantification and (i) representative images of Plp-EGFP OPCs treated with DMSO, T3, or increasing concentrations of the specific LRP1 inhibitor, RAP (One-way ANOVA with Dunnett’s multiple comparisons test, DMSO vs T3 **p=0.0072; conditions plated in sextuplicate; error bars represent +/− SEM; scale bar 150μm).

Article Snippet: Primary antibodies used for immunoblotting were against LRP1 (L2170, 1:1000, Sigma), MBP (SMI-94, 1:1000, Covance), CNP (CNP, 1:2000, Aves Labs), actin (A2228, 1:5000, Sigma).

Techniques: Expressing, Cell Culture

Journal: Frontiers in cellular neuroscience

Article Title: Neural Stem Cells Overexpressing Nerve Growth Factor Improve Functional Recovery in Rats Following Spinal Cord Injury via Modulating Microenvironment and Enhancing Endogenous Neurogenesis.

doi: 10.3389/fncel.2021.773375

Figure Lengend Snippet: FIGURE 1 | Establishment of NGF-modified NSCs and quality evaluation. (A) The design of lentiviral vectors expressing NGF (pLVX-IRES-ZsGreen-rat NGF). (B) NGF-modified NSCs highly expressed nestin and they had no immunoactivity with GFAP, CNPase, and β-tubulin3. (C) RT-PCR showed the mRNA level of NGF was significantly enhanced in NGF-NSCs. (D) Western blotting further confirmed the elevated protein concentration of NGF in NGF-NSCs. (E) Representative immunocytochemical images demonstrated high levels of NGF in NGF-NSCs neurospheres. (F) NGF-NSCs could differentiate into β-tubulin3-positive neurons, GFAP-positive astrocytes, and CNPase-positive oligodendrocytes. Scale bar = 20 µm. **p < 0.01 and ***p < 0.001.

Article Snippet: Following the blocking by normal goat serum, primary antibodies of Nestin (1:100, Proteintech, Wuhan, China), CNPase (1:200, Proteintech), GFAP (1:200, Cell Signaling Technology, Shanghai, China), and β-tubulin3 (1:100, Cell Signaling Technology) were added to the samples and incubated overnight at 4◦C.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Protein Concentration

Journal: Frontiers in cellular neuroscience

Article Title: Neural Stem Cells Overexpressing Nerve Growth Factor Improve Functional Recovery in Rats Following Spinal Cord Injury via Modulating Microenvironment and Enhancing Endogenous Neurogenesis.

doi: 10.3389/fncel.2021.773375

Figure Lengend Snippet: FIGURE 3 | Survival and differentiation of NGF-modified NSCs after transplantation and high levels of NGF at the lesion core. (A) Four weeks after NGF-NSCs transplantation, we observed that NGF + /ZsGreen + cells aggregated in the epicenter of the injured spinal cord, and several of them migrated into the peri-lesion area. In comparison, the sham and control rats did not show a positive signal of ZsGreen, and their immunoactivity of NGF was much weaker. (B) The transplanted NGF-NSCs marked by ZsGreen expressed β-tubulins3 and CNPase, which indicated the NGF-NSCs could differentiate into neurons and oligodendrocytes. (C,D) The western blotting analysis demonstrated that NGF’s protein level was much higher in the NGF-NSCs groups at day 14, 21, 28, and 35 after injury. (E) Similarly, the mRNA level of NGF exhibited the same elevation in the three groups. Scale bar, white bar 5 µm; yellow bar 200 µm. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001.

Article Snippet: Following the blocking by normal goat serum, primary antibodies of Nestin (1:100, Proteintech, Wuhan, China), CNPase (1:200, Proteintech), GFAP (1:200, Cell Signaling Technology, Shanghai, China), and β-tubulin3 (1:100, Cell Signaling Technology) were added to the samples and incubated overnight at 4◦C.

Techniques: Transplantation Assay, Comparison, Control, Western Blot

Journal: Frontiers in cellular neuroscience

Article Title: Neural Stem Cells Overexpressing Nerve Growth Factor Improve Functional Recovery in Rats Following Spinal Cord Injury via Modulating Microenvironment and Enhancing Endogenous Neurogenesis.

doi: 10.3389/fncel.2021.773375

Figure Lengend Snippet: FIGURE 4 | NGF-modified NSCs transplantation reduced the oligodendrocyte loss, attenuated astrocytosis, and demyelination in the peri-lesion segments post SCI. (A) Five weeks after SCI, the sham rats exhibited significantly higher expression of GFAP in the landscape, anterior horn, and around the central canal in the peri-lesion segments (3 mm caudal to the lesion epicenter). The graft of NGF-NSCs dramatically attenuated the abnormal GFAP expression in the same regions. (B) NGF-NSCs treatment reversed the down-regulation of CNPase in the peri-lesion segments (3 mm caudal to the lesion epicenter). after SCI. (C) The representative electoral microscopy of the white matter in the lesion epicenter showed that the integrity of the nerve sheath and corresponding axons were preserved after NGF-NSCs transplantation. Scale bar: white 200 µm; yellow 20 µm. *p < 0.05 and **p < 0.01.

Article Snippet: Following the blocking by normal goat serum, primary antibodies of Nestin (1:100, Proteintech, Wuhan, China), CNPase (1:200, Proteintech), GFAP (1:200, Cell Signaling Technology, Shanghai, China), and β-tubulin3 (1:100, Cell Signaling Technology) were added to the samples and incubated overnight at 4◦C.

Techniques: Transplantation Assay, Expressing, Microscopy