Skip to main content
. 2022 May 4;13(5):435. doi: 10.1038/s41419-022-04886-7

Fig. 4. Knockdown of KLF4 induced senescence in AECs.

Fig. 4

MLE-12 cells were treated with 10 μg/mL bleomycin in culture medium for 12, 24, 48 and 72 h. A At 72 h, MLE-12 cells became enlarged, vacuolated, and sometimes appear with multiple or enlarged nuclei and β-gal stain was preformed to mark senescent cells. B, C Western blotting was performed to evaluate the expression of P21, KLF4 and TERT expression in bleomycin group and control group (B BEAS-2B cells; C MLE-12 cells). D MLE-12 cells were transfected with KLF4 siRNAs or control siRNA (100 nM). Western blotting was performed to evaluate the expression of P21, KLF4 and TERT expression in siRNA group and control group. E KLF4 expression level was assessed with qRT-PCR. F Telomere lengths were tested after constant stimulation of bleomycin in siRNA and control group. G β-gal stain was preformed to mark senescent cells after 72 h stimulation of bleomycin in siRNA group and control group. H Double-labeled immunofluorescent staining were performed to examine the expression of SP-C (green) and TERT (red) in these two groups. *P < 0.05, **P < 0.01, ***P < 0.001 by t-test.