sphk1 activity assay kit  (Abnova)

Journal: Inflammation

Article Title: Blocking SphK1/S1P/S1PR1 Signaling Pathway Alleviates Lung Injury Caused by Sepsis in Acute Ethanol Intoxication Mice

doi: 10.1007/s10753-021-01490-3

Figure Lengend Snippet: PF-543 blocks the SphK1/S1P/S1PR1 signaling pathway. The SphK1 activity ( A ) and S1P level ( B ) in lung tissues were detected by respective kit. C The protein expression of SphK1 and S1PR1 in lung tissues was determined by Western blot. D The mRNA expression of SphK1 and S1PR1 in lung tissues was determined by RT-qPCR analysis. *** P <0.001 vs. sham group. # P <0.05, ## P <0.01 and ### P <0.001 vs. EtOH group. ∆ P <0.05, ∆∆ P<0.01 and ∆∆∆ P <0.001 vs. CLP group. $$$ P <0.001 vs. EtOH+CLP group.

Article Snippet: According to the instructions of the SPHK1 Activity Assay kit (cat. no. KA0906; Abnova) and S1P ELISA kit (cat. no. K-1900; Echelon Biosciences), the substrate required for enzyme reaction was added to the supernatant, which was incubated at 30°C for 3 min, and the levels of SphK1 and S1P were detected by a microplate reader.

Techniques: Activity Assay, Expressing, Western Blot, Quantitative RT-PCR

Journal: Diabetes, Metabolic Syndrome and Obesity: Targets and Therapy

Article Title: Resveratrol Inhibits Lipopolysaccharide-Induced Extracellular Matrix Accumulation and Inflammation in Rat Glomerular Mesangial Cells by SphK1/S1P2/NF-κB Pathway

doi: 10.2147/DMSO.S278267

Figure Lengend Snippet: Effects of RSV on SphK1/S1P2 pathway in LPS-induced RMCs’ proliferation. RMCs were treated with the same method as above. ( A ) RMCs were stimulated by LPS (100 ng/mL) for different periods of time, from 0~48 h, and then the expression of SphK1 and S1P2 was measured by Western blot. ( B ) RMCs were stimulated with LPS for 24 h with or without RSV at 20 μM, SK-II or JTE-013. Then the expression of SphK1 and S1P2 were examined. ( C and D ) SphK1 activity and S1P content were measured in LPS-induced RMCs for 24 h with or without different concentrations of RSV, SK-II or JTE-013 by SphK activity assay kit and ELISA kit, respectively. ( E ) RMCs were stimulated with LPS for 24 h with or without RSV at 20 μM, SK-II or JTE-013. Then FN and ICAM-1 expression were examined. ( F ) RMCs were stimulated with LPS for 12 h with or without RSV at 20 μM, SK-II or JTE-013. Then iNOS expression was examined. Experiments were performed in triplicate with similar results. Data are means ± SEM. # P< 0.05 vs Control, ## P< 0.01 vs Control; * P< 0.05 vs LPS-treated group, ** P <0.01 vs LPS-treated group.

Article Snippet: Both SphK1 activity kit and S1P ELISA kit were from Echelon (Salt Lake City, CT, USA).

Techniques: Expressing, Western Blot, Activity Assay, Enzyme-linked Immunosorbent Assay

Journal: Diabetes, Metabolic Syndrome and Obesity: Targets and Therapy

Article Title: Resveratrol Inhibits Lipopolysaccharide-Induced Extracellular Matrix Accumulation and Inflammation in Rat Glomerular Mesangial Cells by SphK1/S1P2/NF-κB Pathway

doi: 10.2147/DMSO.S278267

Figure Lengend Snippet: Schematic representation of the proposed underlying mechanism leading to renoprotective effect of RSV against LPS-mediated inflammation and mesangial cell proliferation in DN. This diagram summarizes our current study. RSV suppresses SphK1/S1P2 pathway activation in LPS-induced RMCs, which further reduces NF-κB activity. The inhibitory effects of RSV on SphK1/S1P2/NF-κB pathway may result from specific inhibition toward SphK1 and S1P2, then further inhibiting NF-κB pathway, thereby eventually exerting the renoprotective effects on DN, which is independent of its hypoglycemic effect.

Article Snippet: Both SphK1 activity kit and S1P ELISA kit were from Echelon (Salt Lake City, CT, USA).

Techniques: Activation Assay, Activity Assay, Inhibition