Genome-Wide siRNA-Based Functional Genomics of Pigmentation Identifies Novel Genes and Pathways That Impact Melanogenesis in Human Cells (original) (raw)
Figure 1
Validation of novel gene products supporting melanogensis.
A) MNT-1 cells were transfected with the indicated siRNA pools (50 nM final concentration) targeting 35 of the 94 positive regulators of melanogenesis identified in the primary screen. siRNAs targeting Ker7, a gene that does not impact pigment production, were used as a negative control (black bar). A normalized percent inhibition calculation [26] was employed to compare the consequences of each siRNA pool on pigmentation with that observed upon depletion of tyrosinase. Bars represent mean and s.e.m. for n = 3. Red bars indicate failure to significantly suppress pigmentation. The results of the analysis of these 35 genes are shown in this panel (those genes that were not putative autophagy regulators) and in Figure 3 (putative autophagy regulators). A summary of the results for all 35 genes is shown in Table S4. B) A light micrograph of a representative opaque-walled, clear-bottomed 96-well microtiter plate containing MNT-1 cell monolayers 7 days post transfection with the indicated siRNAs is shown. C) Four independent siRNAs targeting the indicated genes (see Table S3 for siRNA sequence information) were separately tested for the capacity to suppress pigmentation as in (A). Associated p-values (student's t-test) are reported in Table S4.