Fig. 4: FOXM1 underpins holoclone-forming cells. | Nature Communications

Fig. 4: FOXM1 underpins holoclone-forming cells.

From: Single-keratinocyte transcriptomic analyses identify different clonal types and proliferative potential mediated by FOXM1 in human epidermal stem cells

Fig. 4

a Western analysis of total cell extracts from cultures generated by holoclones, meroclones, and paraclones isolated by clonal analysis (see “Methods” and Fig. 1a) of sub-confluent normal human primary keratinocytes. Clonal conversion is marked by progressive decrease of FOXM1, p63, and survivin. β1 and β4 integrins remain stable in holoclones and meroclones and are downregulated in paraclones. 14-3-3σ progressively increases during clonal conversion (representative images of n = 3). b Western analysis of total cell extracts from keratinocytes transduced with the indicated shRNA shows that p63 and survivin are almost undetectable in FOXM1-depleted keratinocytes (representative images of n = 3). c Left: clonal analysis of shRNA-transduced clonogenic keratinocytes (see “Methods”) from cultures initiated from three different biopsies of healthy skin (K5, K52, and K71). The percentage of holoclones, meroclones, and paraclones is indicated in red, light blue, and gray columns, respectively (n = 346 clones analyzed). Right: representative cultures generated by these transduced clones. d Representative image of immunofluorescence analysis of FOXM1 in 5-day colonies derived from control (CNT) and FOXM1-transduced keratinocytes. Scale bars, 20 μm. Representative images of three independent experiments are shown. e Western analysis of total cell extracts from cultures generated by control and FOXM1-C or B-transduced keratinocytes shows that p63 and survivin increase after FOXM1 overexpression (representative images of n = 3). f Left: clonal analysis of CNT and FOXM1-transduced clonogenic keratinocytes (see “Methods”) performed at passage 3 (p3) and at passage 6 (p6) and 8 (p8) of cultures initiated from three different biopsies of healthy skin (K38, K49, and K57). The percentage of holoclones, meroclones, and paraclones is indicated in red, light blue, and gray columns, respectively (n = 253 clones analyzed). Right: representative cultures generated by these transduced clones.

Back to article page