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. 2022 Jun 3;13(6):523. doi: 10.1038/s41419-022-04980-w

Fig. 5. Activation of the STING/IRF3/IFN-β pathway in osteosarcoma cells by the SGLT2 inhibitor.

Fig. 5

RT-PCR (a) and Western blot (b) analyses of the mRNA and protein expression levels of specific genes in MG-63 and MNNG/HOS cell lines with or without treatment with SGLT2 inhibitor (canagliflozin, 1 uM) for 48 h. GAPDH served as an internal reference. c TIMER web tool was used to search the relationship of the mRNA expression level of IFNB1 and the infiltration of CD4+/CD8+ T cells in osteosarcoma. RT-PCR (d) and ELISA (e) analyses of the mRNA and protein expression levels of IFN-β in MG-63 and MNNG/HOS cell lines with or without treatment with SGLT2 inhibitor (canagliflozin, 1 uM) for 48 h. Data are presented as the mean ± SD of three independent experiments (***p < 0.001). f ELISA analysis of the protein expression level of IFN-β in MNNG/HOS cell lines with or without treatment with SGLT2 inhibitor (canagliflozin, 0, 0.5, 1, or 2 uM) for 48 h. Data are presented as the mean ± SD of three independent experiments (ns not significant; *p < 0.05; ***p < 0.001). g ELISA analysis of the protein expression level of IFN-β in MNNG/HOS cell lines with or without treatment with SGLT2 inhibitor (canagliflozin, 1 uM) for 0, 12, 24 or 48 h. Data are presented as the mean ± SD of three independent experiments (*p < 0.05; **p < 0.01; ***p < 0.001). h Western Blot analyses of the protein expression levels in STING silenced MG-63 and MNNG/HOS cell lines with or without treatment with SGLT2 inhibitor (canagliflozin, 1 uM) for 48 h. RT-PCR (i) and ELISA (j) analyses of the mRNA and protein expression levels of IFN-β in STING silenced MG-63 and MNNG/HOS cell lines with or without treatment with SGLT2 inhibitor (canagliflozin, 1 uM) for 48 h. Data are presented as the mean ± SD of three independent experiments (***p < 0.001).