Fig. 6: LIF promotes epidermal progenitor renewal and is inhibited by SFRP1. | Cell Death & Disease

Fig. 6: LIF promotes epidermal progenitor renewal and is inhibited by SFRP1.

From: Comprehensive secretome profiling and CRISPR screen identifies SFRP1 as a key inhibitor of epidermal progenitor proliferation

Fig. 6

A Quantitative RT-PCR of SFRP1 and LIF mRNA in control vs. SFRP1-depleted keratinocytes across an in vitro differentiation time course (day 0-2-4). SFRP1_201, isoform SFRP1_201; SFRP1, total SFRP1; LIF_201, isoform LIF_201; LIF, total isoforms. Data are mean ± SEM. (n = 4, multiple paired student’s t-test). p values denoted above comparisons. B Immunoblot of LIF protein in control vs SFRP1-depleted primary keratinocytes across an in vitro differentiation time course. Relative intensity of LIF is denoted, using shSFRP1 at day 4 = 1.0 for comparison. C ELISA of LIF in control vs. SFRP1-depleted primary keratinocytes. shControl was set to 1 for comparison. Data are mean ± SEM. (n = 2, two-tailed ratio student’s t-test). D Immunoblot of SFRP1 in primary keratinocytes following control, SFRP1 knockdown (shSFRP1), and/or LIF knockdown (shLIF). Relative intensity of SFRP1 is denoted and normalized to beta tubulin. sgControl was set to 1 for comparison. E ELISA of LIF in primary keratinocytes following control, shSFRP1, and/or shLIF. Data are mean ± SD. shControl was set to 1 for comparison. (n = 3, one-way ANOVA with a Tukey’s HSD post hoc test). F Crystal violet staining of keratinocyte colonies generated following treatment with shControl, shSFRP1, shLIF, and/or recombinant human LIF (rhLIF). G Quantification of colony formation in (F). ShControl was set to 1 for comparison. Data are means ± SD. (n = 6, one-way ANOVA with a Tukey’s HSD post hoc test). H Immunoblot of primary keratinocyte protein lysates following treatment with shControl, shSFRP1, and/or ß-catenin knockdown (shCTNNB1). Relative intensity of SFRP1 and CTNNB1 is denoted and normalized to beta tubulin. shControl was set to 1 for comparison. I ELISA of LIF in primary keratinocytes following control, shSFRP1, and/or shCTNNB1. Data are mean ± SD. shControl was set to 1 for comparison. (n = 2, one way ANOVA with Tukey’s HSD). J Crystal violet staining of keratinocyte colonies generated following treatment with shControl, shSFRP1, and/or shCTNNB1. K Quantification of colony formation in (J). ShControl was set to 1 for comparison. Data are means ± SD. (n = 6, one-way ANOVA with a Tukey’s HSD post hoc test).

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