Journal: Journal of Neuroinflammation

Article Title: Human PMSCs-derived small extracellular vesicles alleviate neuropathic pain through miR-26a-5p/Wnt5a in SNI mice model

doi: 10.1186/s12974-022-02578-9

Figure Lengend Snippet: miR-26a-5p alleviate mechanic allodynia through a non-canonical Wnt signaling pathway. A–C mRNA levels of Ryk(A), CaMKII ( B ), NFAT(C) on day 3, 7 and 14 after surgery. Data are expressed as fold change compared to sham-operated controls. D–F Quantitative RT-PCR was used to assess the mRNA expression level of Ryk(D), CaMKII (E), NFAT(F). G Western blot was used to assess the level of Ryk, CaMKII, NFAT after Foxy-5 reverse and miR-26a-5p rescue. Representative immunoblots and quantification showing Foxy-5 reversed the effect of miR-26a-5p, while intrathecal injection of miR-26a-5p one day after Foxy-5 again rescued the effect of Foxy5. Data are expressed as fold change compared to the sham group. H–J Protein levels of IL-1β ( H ), IL-6 ( I ), and TNF-α ( J ), in the L4–L5 dorsal spinal cord of mice were tested by ELISA. n = 6 mice for each group. Data are represented as mean ± sem. * p < 0.05, ** p < 0.01

Article Snippet: Membranes were blocked with 5% non-fat milk in buffer containing 0.1% Tween-20 (TBST) for 1 h, washed 3 times in TBST, and incubated overnight at 4 ℃ with primary antibodies including rabbit anti-Wnt5a (1:1000 dilution; Proteintech; USA; 55184-1-AP), rabbit anti-Ryk (1:1000 dilution; Proteintech; USA; 22138-1-AP), rabbit anti-CaMKII (1:1000 dilution; Proteintech; USA; 13730-1-AP), rabbit anti-NFAT (1:1000 dilution; Proteintech; US; 22023-1-AP), rabbit anti-GAPDH (1:10,000 dilution; Proteintech; USA; 10494-1-AP).

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Injection, Enzyme-linked Immunosorbent Assay

Journal: Journal of Neuroinflammation

Article Title: Human PMSCs-derived small extracellular vesicles alleviate neuropathic pain through miR-26a-5p/Wnt5a in SNI mice model

doi: 10.1186/s12974-022-02578-9

Figure Lengend Snippet: Human PMSCs derived sEVs alleviate neuropathic pain through its anti-inflammatory effect. miR-26a-5p regulated Wnt5a/Ryk/CaMKII/NFAT, a non-canonical Wnt signaling, partly take part in the analgesia of hPMSCs derived sEVs through down-regulating the IL-1β, IL-6, and TNF-α

Article Snippet: Membranes were blocked with 5% non-fat milk in buffer containing 0.1% Tween-20 (TBST) for 1 h, washed 3 times in TBST, and incubated overnight at 4 ℃ with primary antibodies including rabbit anti-Wnt5a (1:1000 dilution; Proteintech; USA; 55184-1-AP), rabbit anti-Ryk (1:1000 dilution; Proteintech; USA; 22138-1-AP), rabbit anti-CaMKII (1:1000 dilution; Proteintech; USA; 13730-1-AP), rabbit anti-NFAT (1:1000 dilution; Proteintech; US; 22023-1-AP), rabbit anti-GAPDH (1:10,000 dilution; Proteintech; USA; 10494-1-AP).

Techniques: Derivative Assay

Journal: Animals : an Open Access Journal from MDPI

Article Title: Screening and Identification of Differential Ovarian Proteins before and after Induced Ovulation via Seminal Plasma in Bactrian Camels

doi: 10.3390/ani11123512

Figure Lengend Snippet: Differentially expressed proteins in the ovarian tissues of Bactrian camels after SP-induced ovulation.

Article Snippet: The membranes were blocked for 2 h in Tris–HCl buffer containing 5% ( w / v ) non-fat milk powder, then were incubated overnight at 4 °C with the following primary antibodies from mice or rabbits: ACTB (1:5000, Proteintech, China), CAMK2G (1:1000, Proteintech), FST (1:1000, Proteintech), NR5A1 (1:500, Proteintech), PAK1 (1:1000, Santa Cruz, USA), PRL (1:1000, Santa Cruz), STAG2 (1:1000, Santa Cruz), TGFB2 (1:500, Santa Cruz), Wnt2b (1:500, Santa Cruz), GAPDH (1:5000, Proteintech).

Techniques: Expressing

Journal: Animals : an Open Access Journal from MDPI

Article Title: Screening and Identification of Differential Ovarian Proteins before and after Induced Ovulation via Seminal Plasma in Bactrian Camels

doi: 10.3390/ani11123512

Figure Lengend Snippet: Validation of differentially expressed proteins (DEPs). ( A ) Protein expression levels detected in ovarian protein extracts from Bactrian camels via iTRAQ LC-MS/MS. ( B ) Western blot analysis of PAK1, PRL, STAG2, TGFB2, CAMK2G, FST, NR5A1, and Wnt2b in ovarian protein extracts from Bactrian camels. ( C ) Integrated optical density analysis of the Western blots using ImageJ. Data are presented as the mean ± standard deviation; * p < 0.05, ** p < 0.01. ( D ) Venn diagram of the DEPs using multiple comparisons of the endocrine system, reproductive system and neural signal transduction system groups. ( E ) Immunofluorescence staining to localize the expression patterns of the DEPs in the ovarian samples.

Article Snippet: The membranes were blocked for 2 h in Tris–HCl buffer containing 5% ( w / v ) non-fat milk powder, then were incubated overnight at 4 °C with the following primary antibodies from mice or rabbits: ACTB (1:5000, Proteintech, China), CAMK2G (1:1000, Proteintech), FST (1:1000, Proteintech), NR5A1 (1:500, Proteintech), PAK1 (1:1000, Santa Cruz, USA), PRL (1:1000, Santa Cruz), STAG2 (1:1000, Santa Cruz), TGFB2 (1:500, Santa Cruz), Wnt2b (1:500, Santa Cruz), GAPDH (1:5000, Proteintech).

Techniques: Expressing, Liquid Chromatography with Mass Spectroscopy, Western Blot, Standard Deviation, Transduction, Immunofluorescence, Staining