Tyrosinase Antibody - #AF5491

Figure 4. P4HB regulates the ubiquitination degradation of IRF1. A. Changes in TYR activity in P4HB overexpression or knockdown A375 cells. B. Changes in TYR mRNA expression in Sal treated, P4HB overexpression, and P4HB knockdown A375 cells. C. Western blot analysis of TYR expression in P4HB overexpression or knockdown A375 cells. D. Proteins have interactions with P4HB and USF1 analyzed by FpClass. E. Interaction of P4HB and IRF1 in A375 cells detected by PLA. F. Western blot analysis of the expression of IRF1 and P4HB in P4HB overexpression or knockdown A375 cells. G. Western blot analysis of IRF1 expression in the nucleus of A375 cells after P4HB overexpression or knockdown. H. Effect of SAL on the ubiquitination of IRF1 as determined by Western blot. I. Western blot analysis of IRF1 expression in whole cells and nucleus of SAL-treated A375 cells. Data are expressed as mean ± SD (*P < 0.05, **P < 0.01).

Figure 4 Anti-melanogenic effect of THC on the B16F10 cells. (A) Effects of THC on the viability of B16F10 melanoma cells after 48 h treatment. (B) Melanin content comparison between control group, cells treated with 0.5 μM MSH, and 0.5 μM α-MSH with 1 μg/mL and 10 μg/mL THC, respectively after B16F10 cells were cultured for 48 h. (C) Cellular tyrosinase (TYR) activity. (D) Western blotting analysis of protein expressions for TYR, tyrosinase related protein 1 (TRP1), tyrosinase related protein 2 (TRP2), and Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as the internal reference in B16F10. Results are expressed as mean SD (n = 3). *** p < 0.001 compared to group treated with 0.5 μM α-MSH.

Figure 4 ER retention of variant TYR as the cause of functional loss.A, diagram of TYR::GFP fusion proteins. B, the melanin production of fusion proteins. The fusion vectors were respectively transfected in 293T cells, without l-Tyr treatment. The melanin production was visualized by the color of cell pellets. The scale bars represent 75 μm. C, subcellular localization of translated fusion TYR among organelles. The fusion vectors were cotransfected with ER-dsRed and Golgi-dsRed reporter vectors, respectively, in 293T cells. After 48 h, cells were fixed and observed under the confocal microscopy. The scale bars represent 10 μm. D, 293T cells were transfected with 500 ng fusion vectors, respectively, in a 6-well plate. After 48 h, cells were harvested for Endo H digestion. The digested proteins were separated by Western blot. E and F, 293T cells were transfected with 1 μg fusion vectors, respectively. After 24 h, cells were treated with 100 μg/ml CHX and collected respectively at desired time points (0, 1.5, 3, 6, 12, and 24 h). The half-lives of fusion proteins were measured by Western blot. CHX, cycloheximide; Endo H, endoglycosidase H; ER, endoplasmic reticulum; l-Tyr, l-tyrosine; SP, signal peptide; TYR, tyrosinase.