kca3 1 inhibitor  (MedChemExpress)

Journal: Journal of Inflammation Research

Article Title: KCa3.1 Inhibition of Macrophages Suppresses Inflammatory Response Leading to Endothelial Damage in a Cell Model of Kawasaki Disease

doi: 10.2147/JIR.S297131

Figure Lengend Snippet: Effects of KCa3.1 inhibition by TRAM-34 on LCWE-induced cytotoxicity and activation in RAW264.7 cells. ( A ) RAW264.7 cells were stimulated with LCWE (1 μg/mL) for 12 h. The protein level of KCa3.1 was detected by Western blot. Results are expressed as the mean ± SD. ** p < 0.01 versus control group. ( B , C ) Cytotoxicity of RAW264.7 cells treated by TRAM-34 (0, 1, 5, 10, 15 μM) with or without LCWE (1 μg/mL) was determined by CCK8 assay. Results (n = 5) are expressed as the mean ± SD, * p < 0.05, ** p < 0.01 versus untreated group, # p < 0.05 versus LCWE-treated group. ( D ) The representative microphotographs showed phenotypic changes of RAW 264.7 cells in responsive to different stimuli (LCWE (1 μg/mL), TRAM-34 (10 μM)) for 12 h. Magnification = 100× or 200×. Scale bar = 100 µm.

Article Snippet: KCa3.1 inhibitor TRAM-34, NF-κB inhibitor PDTC and STAT3 inhibitor AG490 were purchased from MCE (Shanghai, China).

Techniques: Inhibition, Activation Assay, Western Blot, Control, CCK-8 Assay

Journal: Journal of Inflammation Research

Article Title: KCa3.1 Inhibition of Macrophages Suppresses Inflammatory Response Leading to Endothelial Damage in a Cell Model of Kawasaki Disease

doi: 10.2147/JIR.S297131

Figure Lengend Snippet: Blocking KCa3.1 with TRAM-34 inhibits LCWE-induced inflammatory response in RAW264.7 cells. RAW264.7 cells were pretreated with TRAM-34 (0, 1, 5, 10 μM) 30 min, then exposed to LCWE (1 μg/mL) for 12 h. The mRNA expressions of IL-1β ( A ), IL-6 ( B ), TNF-α ( C ), MCP-1 ( D ), iNOS ( E ) and MMP-9 ( F ) were assayed by qRT-PCR, respectively. The protein levels of iNOS ( G ) as well as MMP-9 ( I ) were detected using Western blot. ( H ) Culture supernatants of RAW264.7 cells were collected for NO detection by Griess reagent. All results are expressed as the mean ± SD. *** p < 0.001, **** p < 0.0001 versus untreated group; # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001 versus LCWE group.

Article Snippet: KCa3.1 inhibitor TRAM-34, NF-κB inhibitor PDTC and STAT3 inhibitor AG490 were purchased from MCE (Shanghai, China).

Techniques: Blocking Assay, Quantitative RT-PCR, Western Blot

Journal: Journal of Inflammation Research

Article Title: KCa3.1 Inhibition of Macrophages Suppresses Inflammatory Response Leading to Endothelial Damage in a Cell Model of Kawasaki Disease

doi: 10.2147/JIR.S297131

Figure Lengend Snippet: KCa3.1 inhibition by TRAM-34 suppresses inflammatory reaction in LCWE-stimulated mouse peritoneal macrophages. Mouse peritoneal macrophages were pretreated with TRAM-34 (10 μM) 30 min, then stimulated with LCWE (1 μg/mL) for 12 h. The expressions of IL-1β ( A ), IL-6 ( B ), TNF-α ( C ), MCP-1 ( D ), iNOS ( E ) and MMP-9 ( F ) at the mRNA levels were measured using qRT-PCR, respectively. Effects of TRAM-34 pretreatment on the protein expressions of iNOS ( G ) and MMP-9 ( H ) were analyzed by Western blot. All results are expressed as the mean ± SD. ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus control group; # p < 0.05, ## p < 0.01, ### p < 0.001 versus LCWE group.

Article Snippet: KCa3.1 inhibitor TRAM-34, NF-κB inhibitor PDTC and STAT3 inhibitor AG490 were purchased from MCE (Shanghai, China).

Techniques: Inhibition, Quantitative RT-PCR, Western Blot, Control

Journal: Journal of Inflammation Research

Article Title: KCa3.1 Inhibition of Macrophages Suppresses Inflammatory Response Leading to Endothelial Damage in a Cell Model of Kawasaki Disease

doi: 10.2147/JIR.S297131

Figure Lengend Snippet: KCa3.1 blockade by TRAM-34 inhibits the activation of NF-κB and STAT3 caused by LCWE stimulation in RAW264.7 cells. RAW264.7 cells were stimulated with LCWE (1 μg/mL) for 12 h after 30 min pretreatment of TRAM-34 (0, 1, 5, 10 μM). ( A ) The nuclear or cytoplasmic protein levels of NF-κB p65 were detected by Western blot. ( B ) The expressions of p-STAT3 and STAT3 were examined via Western blot. ( C ) The protein levels of p-p38 and p38 were assayed by Western blot. All results are expressed as the mean ± SD. * p < 0.01, ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus untreated group; # p < 0.05, ## p < 0.01, ### p < 0.001 versus LCWE group.

Article Snippet: KCa3.1 inhibitor TRAM-34, NF-κB inhibitor PDTC and STAT3 inhibitor AG490 were purchased from MCE (Shanghai, China).

Techniques: Activation Assay, Western Blot

Journal: Journal of Inflammation Research

Article Title: KCa3.1 Inhibition of Macrophages Suppresses Inflammatory Response Leading to Endothelial Damage in a Cell Model of Kawasaki Disease

doi: 10.2147/JIR.S297131

Figure Lengend Snippet: KCa3.1 inhibition of macrophages alleviates MCAECs inflammation induced by LCWE-treated RAW-CM. MCAECs were incubated with RAW264.7 cells-conditioned medium from various groups (Control, LCWE, LCWE+TRAM-34, LCWE+PDTC, LCWE+AG490) for 12 h. Expressions of IL-1β ( A ), TNF-α ( B ), MCP-1 ( C ) at the mRNA levels was examined by qRT-PCR. All results are expressed as the mean ± SD. *** p < 0.001, **** p < 0.0001 versus control group; # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001 versus LCWE group.

Article Snippet: KCa3.1 inhibitor TRAM-34, NF-κB inhibitor PDTC and STAT3 inhibitor AG490 were purchased from MCE (Shanghai, China).

Techniques: Inhibition, Incubation, Control, Quantitative RT-PCR

Journal: Journal of Inflammation Research

Article Title: KCa3.1 Inhibition of Macrophages Suppresses Inflammatory Response Leading to Endothelial Damage in a Cell Model of Kawasaki Disease

doi: 10.2147/JIR.S297131

Figure Lengend Snippet: KCa3.1 inhibition of macrophages attenuates MCAECs injury mediated by LCWE-treated RAW-CM. MCAECs were incubated with RAW264.7 cells-conditioned medium from various groups (Control, LCWE, LCWE+TRAM-34, LCWE+PDTC, LCWE+AG490) 12 h for the detection of cell viability, MCAECs injury-related protein and STAT3 phosphorylation, or 1 h for the detection of phosphorylation of NF-κB p65. ( A ) Cell survival of MCAECs was determined by using CCK8 assay (n=5). ( B ) Expressions of VCAM-1, MMP-9, IL-1β and KCa3.1 were analyzed by Western blot. ( C ) Expressions of p-p65 and p65 at protein levels were measured by Western blot. ( D ) The protein levels of p-STAT3 and STAT3 were detected by Western blot. All results are expressed as the mean ± SD. *** p < 0.001, **** p < 0.0001 versus control group; # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001 versus LCWE group.

Article Snippet: KCa3.1 inhibitor TRAM-34, NF-κB inhibitor PDTC and STAT3 inhibitor AG490 were purchased from MCE (Shanghai, China).

Techniques: Inhibition, Incubation, Control, CCK-8 Assay, Western Blot