Relative quantitation of the distribution of Pmel17 and TRP1, Pmel17 and EEA1, or Pmel17, TRP1, and LAMP1 was performed on double or triple immunogold-labeled cryosections by counting the number of gold particles labeling each protein in each of the defined compartments taken randomly in 30 cell profiles. Melanosome stages were defined by morphology (see Results), lysosomes were defined by morphology and/or dense labeling for LAMP1, and early endosomes were defined as tubulovesicular structures labeling densely for EEA1. Results are presented as a percentage of the total number of gold particles for each protein in each compartment and represent a mean and standard deviation of two independent experiments.
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Quantitative Analysis of Melanosome Proteins
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Variable analysis
independent variables
- Relative quantitation of the distribution of Pmel17 and TRP1
- Relative quantitation of the distribution of Pmel17 and EEA1
- Relative quantitation of the distribution of Pmel17, TRP1, and LAMP1
- Number of gold particles labeling each protein in each of the defined compartments
- Melanosome stages were defined by morphology
- Lysosomes were defined by morphology and/or dense labeling for LAMP1
- Early endosomes were defined as tubulovesicular structures labeling densely for EEA1
- No positive or negative controls were explicitly mentioned in the provided information.
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