Journal: Neural Regeneration Research

Article Title: Photobiomodulation inhibits the expression of chondroitin sulfate proteoglycans after spinal cord injury via the Sox9 pathway

doi: 10.4103/1673-5374.374136

Figure Lengend Snippet: PBM modulates the Smad3/Sox9 and MAPK/Sox9 pathways in inflammation-induced astrocytes. (A, B) Representative western blots of proteins related to the Smad3/Sox9 pathway, Erk/Sox9 pathway, P38/Sox9 pathway, and JNK/Sox9 pathway in astrocytes and quantification of the relative expression levels of p-Smad3, p-Erk, p-P38, p-JNK, Sox9, and versican. Data are shown as mean ± SD ( n = 3 for each group) and were analyzed by multiple repeated measures analysis of variance with least significance difference post hoc analysis. * P < 0.05, *** P < 0.001, **** P < 0.0001. Erk: Extracellular signal-regulated kinase; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; JNK: c-Jun N-terminal kinase.

Article Snippet: The membranes were soaked in 5% skimmed milk for 1 hour at room temperature and incubated overnight at 4°C with the following primary antibodies: anti-mouse glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (1:3000, Proteintech, Wuhan, Hubei, China, Cat# 60004-1-Ig, RRID: AB_2107436), anti-mouse chondroitin sulfate proteoglycans (CSPG) (1:1000, Sigma-Aldrich, St. Louis, MO, USA, Cat# C8035, RRID: AB_476879), anti-mouse versican (1:1000, Thermo Fisher Scientific, Cat# S351-23, RRID: AB_2735409), anti-rabbit Sox9 (1:1000, Abcam, Cambridge, UK, Cat# ab185966, RRID: AB_2728660), anti-rabbit p-P38 MAPK (P38) (1:1000, CST, Danvers, MA, USA, Cat# D3F9, RRID: AB_2139682), anti-rabbit P38 (1:1000, CST, Cat# D13E1, RRID: TSC_SD02390), anti-rabbit p-extracellular signal-regulated kinase 1/2 (Erk1/2) (1:1000, CST, Cat# D13.14.4E, RRID: AB_10694057), anti-rabbit Erk1/2 (1:1000, CST, Cat# 137F5, RRID: AB_10693607), anti-rabbit p-Smad3 (1:1000, CST, Cat# C25A9, RRID: AB_2193207), anti-rabbit Smad2/3 (1:1000, CST, Cat# D7G7, RRID: AB_10889933), anti-rabbit p-c-Jun N-terminal kinase (JNK) (1:1000, Proteintech, Cat# 80024-1-RR, RRID: AB_2882943), and anti-rabbit JNK (1:1000, Proteintech, Cat# 66210-1-Ig, RRID: AB_2881601).

Techniques: Western Blot, Expressing

Journal: Neural Regeneration Research

Article Title: Photobiomodulation inhibits the expression of chondroitin sulfate proteoglycans after spinal cord injury via the Sox9 pathway

doi: 10.4103/1673-5374.374136

Figure Lengend Snippet: The Smad3/Sox9 pathway and MAPK/Sox9 pathway are involved in photobiomodulation (PBM) modulation of versican expression in spinal cord injury (SCI) mice. (A–C) Representative western blots of proteins related to the Erk/Sox9 pathway (A), P38/Sox9 pathway (B), and Smad3/Sox9 pathway (C) at 7 dpi and quantification of the levels of p-Smad3, p-Erk, p-P38, Sox9, and versican ( n = 6 mice for each group). Data are shown as mean ± SD and were analyzed by one-way analysis of variance with least significance difference post hoc analysis. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. dpi: Day(s) post-injury; Erk: extracellular signal-regulated kinase; GAPDH: glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: The membranes were soaked in 5% skimmed milk for 1 hour at room temperature and incubated overnight at 4°C with the following primary antibodies: anti-mouse glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (1:3000, Proteintech, Wuhan, Hubei, China, Cat# 60004-1-Ig, RRID: AB_2107436), anti-mouse chondroitin sulfate proteoglycans (CSPG) (1:1000, Sigma-Aldrich, St. Louis, MO, USA, Cat# C8035, RRID: AB_476879), anti-mouse versican (1:1000, Thermo Fisher Scientific, Cat# S351-23, RRID: AB_2735409), anti-rabbit Sox9 (1:1000, Abcam, Cambridge, UK, Cat# ab185966, RRID: AB_2728660), anti-rabbit p-P38 MAPK (P38) (1:1000, CST, Danvers, MA, USA, Cat# D3F9, RRID: AB_2139682), anti-rabbit P38 (1:1000, CST, Cat# D13E1, RRID: TSC_SD02390), anti-rabbit p-extracellular signal-regulated kinase 1/2 (Erk1/2) (1:1000, CST, Cat# D13.14.4E, RRID: AB_10694057), anti-rabbit Erk1/2 (1:1000, CST, Cat# 137F5, RRID: AB_10693607), anti-rabbit p-Smad3 (1:1000, CST, Cat# C25A9, RRID: AB_2193207), anti-rabbit Smad2/3 (1:1000, CST, Cat# D7G7, RRID: AB_10889933), anti-rabbit p-c-Jun N-terminal kinase (JNK) (1:1000, Proteintech, Cat# 80024-1-RR, RRID: AB_2882943), and anti-rabbit JNK (1:1000, Proteintech, Cat# 66210-1-Ig, RRID: AB_2881601).

Techniques: Expressing, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Endurance Exercise-Induced Fgf21 Promotes Skeletal Muscle Fiber Conversion through TGF-β1 and p38 MAPK Signaling Pathway

doi: 10.3390/ijms241411401

Figure Lengend Snippet: TGF-β1 and p38 MAPK signaling pathways regulate Fgf21-induced MyHC expression in C2C12 myotubes. C2C12 cells were treated with inhibitors for 24 h and then infected with LV-Fgf21 for 12 h, continued induction of myotube differentiation. ( A ) C2C12 cells were harvested to detect MyHC I, MyHC IIa, MyHC IIb, TGF-β1, Smad2/3, mmp9, and p-p38 via immunoblot. ( B ) Data are shown in the bar graph as the fold change of normalized proteins of β-actin (mean ± SE). ( C ) The mRNA expression of MyHC I, MyHC IIa, MyHC IIb, MyoD, and MyoG were detected via real-time qPCR. Results are reported in the bar graph as the fold changes of mRNAs normalized to β-actin (mean ± SE). ( D ) Cellular immunofluorescence staining was used to detect the impact of pathway inhibitors on the eMyHC-positive myotubes induced by Fgf21 (Scale bar in 200 μm). Bars mean p ≤ 0.05 analyses followed by one-way ANOVA with post hoc Tukey’s tests. * p < 0.05, ** p < 0.01 vs. DMSO; ## p < 0.01 vs. SB431542; & p < 0.05, && p < 0.01 vs. SB203580.

Article Snippet: After blocking the membrane with 5% non-fat dry milk for 1.5 h, primary antibodies were proceeded overnight at 4 °C and included rabbit anti-Fgf21 (ab171941, Abcam), rabbit anti-MyHC I (22280-1-AP, Proteintech), rabbit anti-MyHC IIa (ab124937, Abcam), rabbit anti-MyHC IIb (20140-1-AP, Proteintech), and mouse anti-MyHC IIx (67299-1-Ig, Proteintech), rabbit anti-TGF-β1 (bs-0086R, Bioss, Beijing, China), rabbit anti-Smad2/3 (PA5-99539, Invitrogen), rabbit anti-p-p38 MAPK (8690, CST, MA, USA), mouse anti-Tubulin (T6199, Sigma-Aldrich, MO, USA), and mouse anti-β-actin (4967, CST).

Techniques: Expressing, Infection, Western Blot, Immunofluorescence, Staining

Journal: iScience

Article Title: Targeting endometrial inflammation in intrauterine adhesion ameliorates endometrial fibrosis by priming MSCs to secrete C1INH

doi: 10.1016/j.isci.2023.107201

Figure Lengend Snippet:

Article Snippet: Rabbit monoclonal anti-p-p38 , Cell Signaling Technology , Cat# 9211; RRID: AB_331641.

Techniques: Staining, Recombinant, Cell Counting, Enzyme-linked Immunosorbent Assay, Western Blot, Software