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. Author manuscript; available in PMC: 2022 Jan 1.
Published in final edited form as: J Invest Dermatol. 2020 Jun 30;141(1):152–163. doi: 10.1016/j.jid.2020.05.114

Figure 2. Phenformin enhances human and mouse keratinocyte differentiation induced by suspension assays in vitro.

Figure 2.

a-d: Human keratinocytes cultured in suspension treated with 1 mM phenformin (Phen) or vehicle were collected at the indicated time points for qRT-PCR analysis of keratinocyte differentiation markers CK5, CK1, CK10 and loricrin as indicated. Expression levels of different markers were calculated relative to the control cells at 2 h in suspension. e: Lysates from human keratinocytes treated as in a-d were analyzed for expression of the markers indicated by western blot. f: Quantification of band densities of samples in e from 12 h after suspension. g: Fresh mouse keratinocytes cultured in suspension with phenformin or vehicle were collected for qRT-PCR analysis. h-i: Human keratinocytes were transfected with siRNAs for AMPK α1 and α2 (siAMPKα) or a scrambled siRNA as a control. Cells were collected for analyses by qRT-PCR (h) and western blot (i) for the indicated mRNAs and proteins after 24 h of treatment with 1 mM phenformin or vehicle. ** P<0.01, *** P<0.005, **** P<0.001 by Student’s two tailed t-test.