Figure 11. BECN1 and/or ATG5 depletion reduces the percentage of KRT1-positive keratinocytes upon FGF7 treatment. HaCaT cells were coinjected with EGFP-LC3 cDNA and ATG5 siRNA or BECN1 siRNA or both, to obtain specific gene silencing. Microinjection with an unrelated siRNA was performed as control. After injection, cells were serum-starved or treated with FGF7 as above. Quantitative immunofluorescence analysis shows that in cells microinjected (asterisks) with ATG5 siRNA, with BECN1 siRNA or with ATG5 and BECN1 siRNAs mixture, the increase of the percentage of LC3-positive autophagosomes per cell as well as the expression of KRT1 in response to FGF7 appeared significantly reduced compared with the surrounding uninjected cells in the same microscopic fields or to cells injected with control siRNA. The quantitative analysis was assessed as reported in Materials and Methods, results are expressed as mean values ± standard errors (SE). Student t test was performed and significance level has been defined as P < 0.05: *P < 0,001 vs. the corresponding control siRNA-injected cells. Scale bar: 10 μm.