siRNA-mediated downregulation of HMB45 in differentiated ADSCs.

siRNA resulted in the downregulation of HMB45 in undifferentiated ADSCs and differentiated ADSCs.</p

The expression of melanocytic markers in undifferentiated and differentiated ADSCs detected by immunofluorescence microscopy.

The expression levels of HMB45, MITF, MelEM, Mel2, MelanA, MATP, and tyrosinase (TYR) in undifferentiated and differentiated ADSCs, as well as NHEMs, were examined by immunofluorescence microscopy. All melanocytic markers except for TYR were detected.</p

The expression of HMB45 and MITF in Mel2-positive and Mel2-negative cells.

The expression of HMB45 and MITF in Mel2-positive cells was increased compared to that in Mel2-negative cells, as shown by immunofluorescence microscopy (A). The expression of HMB45 and MITF in Mel2-positive cells was increased compared to that in Mel2-negative cells, as shown by RT-PCR (B). There was almost no difference in the expression of KIT in Mel2-positive cells and Mel2-negative cells, as shown by RT-PCR (B). The expression of MITF in Mel2-positive cells was increased compared to that in Mel2-negative cells, as shown by western blot analysis (C).</p

The expression of melanocytic markers in undifferentiated and differentiated ADSCs detected by RT-PCR.

The expression levels of HMB45, MITF, PAX3, and KIT in undifferentiated ADSCs and differentiated ADSCs were compared with those in melanocytes by RT-PCR. The expression levels of HMB45, MITF, PAX3, and KIT were higher in differentiated ADSCs than in undifferentiated ADSCs.</p

Three-dimensional epidermal culture using normal human epidermal keratinocytes (NHEKs) and normal human epidermal melanocytes (NHEMs) and NHEKs and differentiated ADSCs.

HMB45- and tyrosinase (TYR)-positive cells were observed in the basal layer of both epidermis-like structures (A). Melanin deposits were induced by UVB irradiation in 3D epidermal culture using NHEKs and NHEMs, as well as in NHEKs and differentiated ADSCs and were detected by Fontana-Masson staining (B).</p

Immunofluorescence and immunoelectron microscopy with an anti-HMB45 antibody in undifferentiated ADSCs, differentiated ADSCs and melanoma cells.

Note that the immunostaining intensity was increased in differentiated ADSCs. Immunogold particles mainly reacted with tubular structures due to the lack of mature melanosomes.</p

The expression of HMB45 and MITF in undifferentiated and differentiated ADSCs by western blotting.

HMB45 expression in undifferentiated and differentiated ADSCs was detected by western blotting (A). MITF expression in undifferentiated and differentiated ADSCs was detected by western blotting (B). The expression of MITF was increased in differentiated ADSCs.</p

Pigmented cells were detected among differentiated ADSCs in an L-DOPA reaction assay.

An L-DOPA reaction assay was performed on undifferentiated ADSCs, differentiated ADSCs, NHEMs, and Mel2-positive and Mel2-negative cells. Pigmented L-DOPA-positive cells were observed among differentiated ADSCs and NHEMs (A). The L-DOPA reaction was not significantly different in Mel2-positive cells and Mel2-negative cells (B).</p

S1 File -

(DOCX)</p

Comparison of architecture of PL and IL of vmPFC between OXT and PBS groups at P40.

Nissl-stained sections of OXT (A) and PBS (B) groups. Scale bar: 100 μm. (TIF)</p