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. 2020 Oct 22;19:294–307. doi: 10.1016/j.omto.2020.10.009

Figure 1.

Figure 1

Therapeutic Potential of miR-634 Ointment for cSCC

(A) Cell growth assay in 2D or 3D in vitro. Scale bars, 2D, 50 μm; 3D, 100 μm. Bars indicate the SD. The error bars are not visible because they are too small. Data are presented as mean ± SD. p values were calculated using the two-sided Student’s t test (∗p = 0.0001, ∗∗p = 0.002). (B) Western blotting analysis of resected tumors. Cell lysates were separated by SDS-PAGE and immunoreacted with the indicated antibodies. (C) Representative images of mitochondrial staining. Images are represented in the grayscale. Scale bars, 2 μm. (D) Western blotting analysis of resected tumors. Cell lysates were separated by SDS-PAGE and immunoreacted with the indicated antibodies. The arrow indicates the band corresponding to LC3B-II, an autophagosome marker. (E) Experimental schedule for therapeutic applications. (F) In vivo tumor growth assay. Upper panel: representative images of subcutaneous tumors at day 6 after initial treatment. Lower left panel: tumor volume in mice treated with miR-NC ointment (n = 7) or miR-634 ointment (n = 7). Errors bars indicate the SD. Lower right panel: scatterplot of tumor weight in mice treated with miR-NC ointment or miR-634 ointment. Data are presented as mean ± SD. p values were calculated using the two-sided Student’s t test (∗p = 0.0017, ∗∗p < 0.0001). (G) Representative images of ISH in tumors from mice treated with miR-NC ointment or miR-634 ointment. The miR-634-specific probe appears purple in the cytoplasm, and the nucleus was counterstained with nuclear fast red. Scale bars, 100 μm. (H) qRT-PCR analysis of miR-634 expression in resected tumors. miR-634 expression levels in tumors from mice treated with miR-NC ointment (n = 4) or miR-634 ointment (n = 4) were measured by qRT-PCR, and the results are shown in the boxplot. P values were calculated using the two-sided Student’s t test (∗p = 0.0326). (I) Western blotting analysis of resected tumors. Cell lysates were separated by SDS-PAGE and immunoreacted with the indicated antibodies. (J) Relative expression levels of target genes. The expression levels of target genes in tumors treated with miR-634 ointment relative to those treated with miR-NC ointment are presented as boxplots. P values were calculated using the two-sided Student’s t test.