antibodies against mitf  (Huabio Inc)

Journal: International Journal of Molecular Medicine

Article Title: Giant Centella asiatica , a novel cultivar rich in madecassoside and asiaticoside, suppresses α-melanocyte-stimulating hormone-induced melanogenesis through MC1R binding

doi: 10.3892/ijmm.2024.5454

Figure Lengend Snippet: Inhibition of the melanogenesis-related signaling and tyrosinase activity by the GCA extract. (A) The GCA extract reduces cAMP levels, indicating its potent modulatory effect on cAMP-dependent pathways critical for melanogenesis. ## P<0.01 between the control and α-MSH. Data are presented as mean values ± SD (n=3). Mean values with different letters indicate statistically significant differences among the treatment groups, including the α-MSH-induced group, as determined by one-way ANOVA followed by Tukey's HSD test (P<0.05). (B) Concentration-dependent inhibitory effect of the GCA extract and arbutin on mushroom tyrosinase activity. Significant differences were observed between the tyrosinase and tyrosinase group + L-DOPA group ( ## P<0.01). Moreover, the addition of GCA at concentrations of 25, 50 and 100 µ g/ml and arbutin 100 µ g/ml to the tyrosinase + L-DOPA group significantly reduced the relative mushroom tyrosinase activity compared with that in the tyrosinase + L-DOPA group, as determined by one-way ANOVA followed by Tukey's HSD test (P<0.05). (C) Effects of GCA extract on the expression and phosphorylation of PKA, CREB, MITF and tyrosinase in B16F10 cells. Protein expression levels were determined in cell lysates using specific antibodies by immunoblotting. β-actin was used as a loading control. Representative western blots from three independent experiments are shown (n=3). Significant differences between untreated control and α-MSH-induced group ( ## P<0.01). Mean values with different letters indicate statistically significant differences among the treatment groups, including the α-MSH-induced group, as determined by one-way ANOVA followed by Tukey's HSD test (P<0.05). α-MSH, α-melanocyte stimulating hormone; CA, Centella asiatica ; GCA, Giant CA; PKA, cAMP-dependent protein kinase; CREB, cAMP response element-binding protein; MITF, microphthalmia-associated transcription factor.

Article Snippet: Antibodies against MITF (cat. no. AB59201), phospho-PKA (p-PKA; cat. no. 4781S), PKA (cat. no. 4782S), p-CREB (cat. no. 9198S) and CREB (cat. no. 9197S) were purchased from Cell Signaling Technology, Inc.

Techniques: Inhibition, Activity Assay, Control, Concentration Assay, Expressing, Phospho-proteomics, Western Blot, Binding Assay

Journal: International Journal of Molecular Sciences

Article Title: Momordica charantia Extract Ameliorates Melanoma Cell Proliferation and Invasion into Mouse Lungs by Suppressing PAX3 Expression

doi: 10.3390/ijms252312800

Figure Lengend Snippet: Effects of MC extract treatment on the expression levels of malignant tumor survival genes located downstream of microphthalmia-associated transcription factor (MITF) in the lungs of melanoma-bearing mice. The MITF ( a , b ), cyclin-dependent kinase 2 (CDK2) ( a , c ), hepatocyte growth factor receptor c-Met ( a , d ), B-cell/CLL lymphoma 2 (Bcl2) ( a , e ), and Ras-related protein (RAB27A) ( a , f ) levels were analyzed. Values are expressed in terms of mean ± SD ( n = 5 animals). Intensity was calculated from five random visual fields with a constant area by using the ImageJ software. Scale bar = 100 μm. * p < 0.05; ** p < 0.01.

Article Snippet: After electrophoresis, the membranes were incubated with primary antibodies against MITF (1:1000; Proteintech Group), CDK2 (1:1000; Abcam), c-Met (1:1000; Abcam), Bcl2 (1:1000; Bioss Inc., Woburn, MA, USA), RAB27A (1:1000; Proteintech Group), and β-actin (1:5000; Sigma-Aldrich, St. Louis, MO, USA) for 1 h at room temperature. β-actin was used as the loading control.

Techniques: Expressing, Software

Journal: International Journal of Molecular Sciences

Article Title: Momordica charantia Extract Ameliorates Melanoma Cell Proliferation and Invasion into Mouse Lungs by Suppressing PAX3 Expression

doi: 10.3390/ijms252312800

Figure Lengend Snippet: Mechanism underlying the inhibition of melanoma cell proliferation and invasion by the MC extract. PAX3 increases the activity of MITF and promotes the proliferation and invasion of melanoma by increasing CDK2, c-MET, Bcl2, and RAB27A. PAX3 also suppresses PTEN and activates signaling from PIP3 to AKT, mTORC1, and S6K1, which promotes melanoma proliferation. The MC extract improves melanoma by suppressing the MITF pathway and PIP3 pathway through the control of PAX3.

Article Snippet: After electrophoresis, the membranes were incubated with primary antibodies against MITF (1:1000; Proteintech Group), CDK2 (1:1000; Abcam), c-Met (1:1000; Abcam), Bcl2 (1:1000; Bioss Inc., Woburn, MA, USA), RAB27A (1:1000; Proteintech Group), and β-actin (1:5000; Sigma-Aldrich, St. Louis, MO, USA) for 1 h at room temperature. β-actin was used as the loading control.

Techniques: Inhibition, Activity Assay, Control

Journal: Cell Death & Disease

Article Title: Transcription factor activating enhancer-binding protein 2ε (AP2ε) modulates phenotypic plasticity and progression of malignant melanoma

doi: 10.1038/s41419-024-06733-3

Figure Lengend Snippet: A Representative Western Blot images depicting murine MITF and BRN2 protein levels in primary AP2ε -/- /Tg(GRM1) and Tg(GRM1) cells (left panels) and human MITF and BRN2 in Mel Juso cells transfected with the AP2ε-overexpression (hAP2ε) plasmid and control vector pCMX (right panels). β-Actin served as loading control. B Immunohistochemical staining of MITF and BRN2 protein expression in Mel Im melanoma cells bioprinted in CIB (Magnification: 20x). Quantification of percentage of cells with “no”, “weak”, or “strong” MITF and BRN2 expression. Data are represented as mean ± SEM; * p < 0.05; ns : not significant (Two-way-ANOVA with Fisher’s LSD test). C mRNA- and protein expression analysis for Snail1 in AP2ε -/- /Tg(GRM1) and Tg(GRM1) cells ( n = 3). D mRNA- and protein expression analysis for Snail1 in Mel Juso cells transfected with the AP2ε-overexpression (hAP2ε) plasmid and control vector pCMX ( n = 3). E mRNA- and protein expression analysis for E-cadherin in AP2ε -/- /Tg(GRM1) and Tg(GRM1) cells ( n = 3). F mRNA- and protein expression analysis for E-cadherin in Mel Juso cells transfected with the AP2ε-overexpression (hAP2ε) plasmid and control vector pCMX ( n = 3). Data information: All data from at least three independent experiments are represented as mean ± SEM; * p < 0.05 (Two-tailed Student’s t-test; ns : not significant).

Article Snippet: Primary antibodies against MITF (1:500 in 5% BSA, Santa Cruz, sc-515925, RRID:AB_2828036), BRN2 (1:500 in 5% BSA/1x TBS-T, Santa Cruz, sc-393324, RRID:AB_2737347), Snail1 (1:1,000 in 5% BSA/1x TBS-T, Cell Signaling Technology Cat# 3879, RRID:AB_2255011) and E-cadherin (1:1,000 in 5% BSA/1x TBS-T, Cell Signaling Technology Cat# 3195, RRID:AB_2291471) were incubated overnight at 4 °C.

Techniques: Western Blot, Transfection, Over Expression, Plasmid Preparation, Control, Immunohistochemical staining, Staining, Expressing, Two Tailed Test