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. 2021 Oct 17;11(20):9918–9936. doi: 10.7150/thno.66378

Figure 6.

Figure 6

Downregulation of melanogenesis by the loss of CRTC3 without affecting melanocyte development. (A) Microscopic images of cultured primary melanocytes from control (CRTL) versus CRTC3 null mice at day 30 and 70 (Bar = 100 µm). (B) Protein level of melanocytic genes including CRTC3, MITF, Tyr, Tyrp1, DCT, PCREB, CREB, Pmel, SOX10 in 70-d cultured primary melanocytes from CTRL and CRTC3 null mice in response FSK for 6 h. (C) Heatmap of significant differentially expressed melanocyte-relevant genes by RNA sequencing experiments in 1-day-old CTRL and CRTC3 null mice tail skin. (D) Protein level of CRTC3, SOX10, MITF, Tyr, Tyrp1, and DCT in CTRL and CRTC3 null mice. α-tubulin was used as an internal loading control. (E) Relative mRNA expression of melanogenesis genes, CRTC3 and SOX10 form 1-day-old CTRL and CRTC3 null mice tail skins (n=4, each) as assessed by qRT-PCR. Immunohistochemistry using SOX10 antibodies in the tail skin of (F) 1-day-old and (G) 2-month-old CTRL and CRTC3 null mice (Bar = 50 µm). (H) Relative mRNA level of SOX10 in tail skin of 2-month-old CTRL and CRTC3 null mice. (I) Protein levels of SOX10 and Pmel in the tail skin from 2-month-old CTRL and CRTC3 null mice. (J) Protein levels of SOX10 and Pmel in control or CRTC3 K/D Mel-ab cells with FSK stimulation for 72 h (K) Immunofluorescence staining using Pmel antibody with nuclear DAPI staining of 2-month-old CTRL and CRTC3 null mice tail skin (Bar = 50 µm).