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. 2007 Sep 14;40(5):685–705. doi: 10.1111/j.1365-2184.2007.00464.x

Figure 3.

Figure 3

Bone morphogenetic factor‐4 (BMP4) in CM and effects of recombinant BMP4 on keratinocyte gene expression. (a) Immunoblot analysis of conditioned (CM) and unconditioned (uCM) media show BMP4 in the CM (24.6 ng/mL). LIF was 2.2‐fold higher in uCM than in CM. Various concentrations of recombinant BMP4 were used as standards by which to quantify the amount of BMP4 present in the CM. 3 mL lyophilized medium loaded per well. (b–g) RT‐PCR; (b) no activation of Oct‐4 or its target transcripts in keratinocytes exposed to 50 ng/mL BMP4 in the presence of FBS. (c) 100 ng/mL of BMP4 activated Oct‐4, Nanog, Utf1, Rex‐1 and Sox‐2 in the presence and (d) absence of FBS. (e) After 5 days exposure to 100 ng/mL BMP4, keratinocytes still expressed Oct‐4, Nanog, Utf1, Rex‐1 and Sox‐2. (f) Keratinocytes cultured in 1 : 1 DKSFM : uCM for 5 days without BMP4 expressed K14. (g) In keratinocytes cultured in 1 : 1 DKSFM : uCM, plus 100 ng/mL BMP4 (renewed daily), for 5 days, K14 was down‐regulated by Day 3, and extinguished by day 5. (f–g) Lane 1 = GAPDH (135 bp); Lane 2 = Keratin 14 (K14) (513 bp).