Journal: Learning & Memory

Article Title: Learning-induced mRNA alterations in olfactory bulb mitral cells in neonatal rats

doi: 10.1101/lm.051177.119

Figure Lengend Snippet: Model of training-induced plasticity changes predicted by up-regulation of transcription in five functionally characterized proteins in the microarray. The protein-coding genes in red support structural changes such as increased connectivity (Dcbdl1, Sec23b), increased synaptic membrane (Sec23b), increased ryanodine receptor clustering (Clic2), increased fatty acid production for mitochondrial biogenesis (Rpp14), and increased excitatory input to granule cells (Mstn).

Article Snippet: The antibodies used were Dcbld1 (1:1500-1:750, Santa Cruz Biotechnology), Clic2 (1:200, Santa Cruz Biotechnology), Sema4c (1:1000-1:500, Aviva Systems Biology), Magee2 (1:500-1:300, Aviva Systems Biology), Scrg1 (1:500, Aviva Systems Biology), and Sec23b (1:1000, Aviva Systems Biology).

Techniques: Microarray, Membrane

Journal: Learning & Memory

Article Title: Learning-induced mRNA alterations in olfactory bulb mitral cells in neonatal rats

doi: 10.1101/lm.051177.119

Figure Lengend Snippet: ( A ) Sec23b immunohistochemical staining in olfactory bulbs ipsilateral to occluded and open nares 50 min following odor + stroking training. ( B 1) Mitral cell layer ROD values for olfactory bulb quadrants receiving input from occluded and open nares. ( B 2) Dorsomedial mitral cell differences observed in the caudal samples for Sec23b. Error bars represent SEMs. * P < 0.05, see results for exact values.

Article Snippet: The antibodies used were Dcbld1 (1:1500-1:750, Santa Cruz Biotechnology), Clic2 (1:200, Santa Cruz Biotechnology), Sema4c (1:1000-1:500, Aviva Systems Biology), Magee2 (1:500-1:300, Aviva Systems Biology), Scrg1 (1:500, Aviva Systems Biology), and Sec23b (1:1000, Aviva Systems Biology).

Techniques: Immunohistochemical staining, Staining

Journal: Stem Cells International

Article Title: Scrapie-Responsive Gene 1 Promotes Chondrogenic Differentiation of Umbilical Cord Mesenchymal Stem Cells via Wnt5a

doi: 10.1155/2022/9124277

Figure Lengend Snippet: Primer sequences used in this study.

Article Snippet: Endogenous peroxidase activity was blocked with 3% H 2 O 2 for 10 min. After blocking with 5% goat serum at room temperature for 1 h, slices were incubated overnight at 4°C with the following primary antibodies: SCRG1 (1 : 800; 14038-1-AP, Proteintech, USA), Col2a1 (1 : 400; 28459-1-AP, Proteintech, USA), and ACAN (1 : 800; 13880-1-AP-100 μ L, Proteintech, USA).

Techniques:

Journal: Stem Cells International

Article Title: Scrapie-Responsive Gene 1 Promotes Chondrogenic Differentiation of Umbilical Cord Mesenchymal Stem Cells via Wnt5a

doi: 10.1155/2022/9124277

Figure Lengend Snippet: SCRG1 was upregulated during chondrogenic differentiation. (a) Alcian Blue staining of UCMSCs chondrogenic differentiation at day 0 and day 21; results showed accumulation of glycosaminoglycans at day 21 after chondrogenic differentiation, indicating chondrogenesis of UCMSCs. Scale bar = 100 μ m. (b) mRNA expression levels of chondrogenic genes ( SOX9 , COL2A1 , and ACAN ) were upregulated during induction. (c) mRNA expression level of SCRG1 was upregulated during induction. (d, e) Protein level of SCRG1 was upregulated after 21 days chondrogenic differentiation, as detected by IHC. Scale bar = 100 μ m. Experiments were repeated three times; ∗ P < 0.05 ( t -test).

Article Snippet: Endogenous peroxidase activity was blocked with 3% H 2 O 2 for 10 min. After blocking with 5% goat serum at room temperature for 1 h, slices were incubated overnight at 4°C with the following primary antibodies: SCRG1 (1 : 800; 14038-1-AP, Proteintech, USA), Col2a1 (1 : 400; 28459-1-AP, Proteintech, USA), and ACAN (1 : 800; 13880-1-AP-100 μ L, Proteintech, USA).

Techniques: Staining, Expressing

Journal: Stem Cells International

Article Title: Scrapie-Responsive Gene 1 Promotes Chondrogenic Differentiation of Umbilical Cord Mesenchymal Stem Cells via Wnt5a

doi: 10.1155/2022/9124277

Figure Lengend Snippet: SCRG1 knockdown inhibited chondrogenic differentiation potential of UCMSCs. (a–c) mRNA and protein expression levels of SCRG1 were decreased after transfection with SCRG1 shRNA, detected by qRT-PCR and IHC. Scale bar = 100 μ m. (d) SOX9 , COL2A1 , and ACAN mRNA levels decreased following SCRG1 knockdown. (e) Alcian Blue staining at day 21. Results showed that glycosaminoglycan synthesis decreased significantly after SCRG1 knockdown. (f, g) Protein levels of COL2A1 and ACAN detected by IHC at day 21 decreased after SCRG1 knockdown. Scale bar = 200 μ m. Experiments were repeated three times; ∗ P < 0.05 ( t -test).

Article Snippet: Endogenous peroxidase activity was blocked with 3% H 2 O 2 for 10 min. After blocking with 5% goat serum at room temperature for 1 h, slices were incubated overnight at 4°C with the following primary antibodies: SCRG1 (1 : 800; 14038-1-AP, Proteintech, USA), Col2a1 (1 : 400; 28459-1-AP, Proteintech, USA), and ACAN (1 : 800; 13880-1-AP-100 μ L, Proteintech, USA).

Techniques: Knockdown, Expressing, Transfection, shRNA, Quantitative RT-PCR, Staining

Journal: Stem Cells International

Article Title: Scrapie-Responsive Gene 1 Promotes Chondrogenic Differentiation of Umbilical Cord Mesenchymal Stem Cells via Wnt5a

doi: 10.1155/2022/9124277

Figure Lengend Snippet: SCRG1 overexpression promoted chondrogenic differentiation of UCMSCs. (a–c) mRNA and protein levels of SCRG1 were detected by qRT-PCR and IHC. Results showed that mRNA and protein levels of SCRG1 increased after overexpression of SCRG1. Scale bar = 100 μ m. (d) SOX9 , COL2A1 , and ACAN mRNA levels increased after overexpression of SCRG1. (e) Alcian Blue staining showed an increase in glycosaminoglycans at day 21 after overexpression of SCRG1. Scale bar = 200 μ m. (f, g) Protein levels of COL2A1 and ACAN detected by IHC at day 21 increased after SCRG1 knockdown. Scale bar = 200 μ m. Experiments were repeated three times; ∗ P < 0.05 ( t -test).

Article Snippet: Endogenous peroxidase activity was blocked with 3% H 2 O 2 for 10 min. After blocking with 5% goat serum at room temperature for 1 h, slices were incubated overnight at 4°C with the following primary antibodies: SCRG1 (1 : 800; 14038-1-AP, Proteintech, USA), Col2a1 (1 : 400; 28459-1-AP, Proteintech, USA), and ACAN (1 : 800; 13880-1-AP-100 μ L, Proteintech, USA).

Techniques: Over Expression, Quantitative RT-PCR, Staining, Knockdown

Journal: Stem Cells International

Article Title: Scrapie-Responsive Gene 1 Promotes Chondrogenic Differentiation of Umbilical Cord Mesenchymal Stem Cells via Wnt5a

doi: 10.1155/2022/9124277

Figure Lengend Snippet: SCRG1 regulated chondrogenesis of UCMSCs through Wnt5a signaling pathway. (a) Regulating pluripotency of stem cell signaling pathway was the most significantly enriched KEGG pathway based on GSEA, wnt5a was negatively correlated with this pathway after SCRG1 overexpression. (b, c) Western blot analysis showed that Wnt5a decreased after shSCRG1 transfection and was restored after administration of rhSCRG1 (450 ng/mL); protein levels of β -catenin exhibited the opposite trend to Wnt5a. (d, e) Protein levels of COL2A1 and ACAN were detected in UCMSCs by IHC. The results showed that the protein levels of COL2A1 and ACAN were reduced following SCRG1 knockdown and restored after treated with rhWnt5a (300 ng/mL) and rhSCRG1 (450 ng/mL) protein at 24 h. Scale bar = 100 μ m. Experiments were repeated three times, ∗ P < 0.05 ( t -test).

Article Snippet: Endogenous peroxidase activity was blocked with 3% H 2 O 2 for 10 min. After blocking with 5% goat serum at room temperature for 1 h, slices were incubated overnight at 4°C with the following primary antibodies: SCRG1 (1 : 800; 14038-1-AP, Proteintech, USA), Col2a1 (1 : 400; 28459-1-AP, Proteintech, USA), and ACAN (1 : 800; 13880-1-AP-100 μ L, Proteintech, USA).

Techniques: Over Expression, Western Blot, Transfection, Knockdown