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. 2022 Feb 4;25(3):103871. doi: 10.1016/j.isci.2022.103871

Figure 4.

Figure 4

Influences of irradiated SZ95 conditioned medium on NHMs melanogenesis and NHFs phenotype

(A) Experimental SZ95 pool medium description and phase-contrast analysis of NHMs treated with S or R UVA 5 J/cm2 for 5 days.

(B) The mRNA expression levels of MITF, TYR, SOX9, and WNT5a in NHMs after addition with S or R UVA 5 J/cm2 for 48h. Results are presented as the mean ± SD of three independent experiments and are expressed as the fold change respect to untreated control cells (∗p < 0.05 vs Ctr).

(C) Western blot analysis of tyr, p53, and p21 protein expression in NHMs after addition with S or R UVA 5 J/cm2 for 72h. GAPDH was used as an equal loading control. Representative blots are shown. Densitometric scanning of band intensities was performed to quantify the change of protein expression (control value taken as 1-fold in each case).

(D) Analysis of tyrosinase activity on NHMs after addition with S or R UVA 5 J/cm2 for 4 days. Results are presented as the mean ± SD of three independent experiments and are expressed as the fold change respect to untreated control cells (∗p < 0.05).

(E) Melanin content evaluation in NHMs after addition with S or R UVA 5 J/cm2 for 5 days. Results are presented as the mean ± SD of three independent experiments and are expressed as ratio of μg melanin/mg protein (∗p < 0.05 versus Ctr).

(F) Experimental SZ95 pool medium description and phase-contrast analysis of NHFs added with S or R UVA 5 J/cm2 for 4 days.

(G) Western blot analysis of α-SMA, p53, and p21 protein expression in NHFs after addition with S or R UVA 5 J/cm2 for 4 days. GAPDH was used as an equal loading control. Representative blots are shown. Densitometric scanning of band intensities was performed to quantify the change of protein expression (control value taken as 1-fold in each case).

(H) The mRNA expression levels of IL-1α, IL-1β, IL-6, IL-8, b-FGF, EDN1, KGF, NRG1, SCF, VEGF, DKK1, WIF1, GDF15, α-SMA, and MMP1 in NHFs after addition with S or R UVA 5 J/cm2 for 48h. Results are presented as the mean ± SD of three independent experiments and are expressed as the fold change respect to untreated control cells (∗p < 0.05, ∗∗p < 0.01 vs Ctr).