Phospho-TMEM173/STING (Ser366) Antibody - #AF7416

Fig. 5. CPC inhibits cGAS-STING pathway activation. (A) Mechanism of CPC inhibiting the activation of cGAS-STING signaling pathway. (B) Immunofluorescence staining and colocalization quantification of DAPI/TOM20/dsDNA in H9C2 cells from different treatment groups. Cytoplasmic dsDNA is indicated by yellow arrow, scale bar: 20 μm. (C) Quantitative real-time PCR analysis of mtDNA: mtDNA content was normalized to nuclear DNA. (D) Determination of the level of cGAMP in H9C2 cells. (E–I) WB of cGAS-STING pathway-related proteins in H9C2 cells, including representative images and quantitative analysis of cGAS, P-STING/STING, P-NF-κB/NF-κB and P-IRF3/IRF3. (J) Determination of the level of IL-1β in H9C2 cells. (K) Determination of the level of IL-6 in H9C2 cells. (L) Immunofluorescence staining of DAPI/TOM20/dsDNA in heart tissue from different treatment groups, the cytoplasmic dsDNA is indicated by yellow arrow, scale bar: 20 μm and 10 μm. (M) The colocalization analysis of DAPI/TOM20/dsDNA in heart tissues: the intensity profiles of the fluorescence signals. (N) Representative images of cGAS and P-STING immunohistochemical staining in heart tissues of different treatment groups, scale bar: 50 μm. Data were expressed as mean ± SE. Statistical significance was performed by one-way ANOVA with Tukey post hoc test. (n = 3, ns: P > 0.05, ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001).

Fig. 5. HPTM with NIR and RT induced accumulation of cytosolic DNA, activation of cGAS-STING and ICD. (A) Representative fluorescence images and (B) quantitative analysis of Picogreen(green) staining of K7M2 cells treated by different treatments, including HPTM, HP + NIR + RT, RT, HPTM + NIR, HPTM + RT, HPTM + NIR + RT. (C) Activation of cGAS-STING related protein was measured by Western Blot. (D) Cytokine levels of IFN-γ in K7M2 cells by ELISA on day 3 post various treatments. (E) Released ATP and (F) HMGB1 were quantified through ATP detection kits and ELISA kits, respectively. (G) Calreticulin immunofluorescence on the outer surface of K7M2RR cells was imaged by LSCM. All experiments were carried out with 3 biological replicates. # indicates a group which was compared with other groups. All values are the mean ± SD, n = 3. P values, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001,∗∗∗∗p < 0.0001. ns, not significant.