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. 2020 Dec 8;10(12):1645. doi: 10.3390/biom10121645

Figure 5.

Figure 5

Effects of BDNF alone or that combined with K252a on the proliferation in Ishikawa cells. Ishikawa cells were monocultured with 100 ng/mL K252a for 30 min and then cocultured with 200 ng/mL BDNF for 24 h. (A) Cell counting kit-8 analysis was performed for the detection of cell viability. (B) Colony formation assay was used in detecting control cell proliferation. (C) Colony formation assay was used in detecting the cell proliferation treated with BDNF. (D) Colony formation assay was used in detecting the cell proliferation treated with K252a. (E) Colony formation assay was used in detecting the cell proliferation treated with K252a and BDNF. (F) Relative mRNA expression levels of CCND1 and CCNE2 genes were detected by qRT-PCR. (G) Protein levels of CCND1 and CCNE2 genes were detected by Western blot analysis. (H) Relative protein levels of CCND1 and CCNE2 genes were analyzed by grey scanning. The expression levels of mRNA and proteins were normalized by GADPH gene, which plays an important role as an internal reference calibration for q RT-PCR and Western blot analysis. Statistical analysis is shown. * p < 0.05, ** p < 0.01.