Fig 1: Effect of APAF1 knockdown on the proliferation and apoptosis of A-431 cells. (A) The expression of miR-186 was examined by reverse transcription-quantitative polymerase chain reaction in A-431 cells with or without si-APAF1 in the presence of miR-186 inhibitors. (B) Western blotting was performed to detect the expression of APAF1, LC3B and Beclin1 proteins in A-431 cells treated with miR-186 inhibitors in the presence or absence of si-APAF1. (C) The invasive ability of A-431 cells treated with miR-186 inhibitors alone or in combination with si-APAF1 was measured using a Matrigel assay. Magnification, ×200. (D) Hoechst 33258 staining was performed to assess the apoptosis of A-431 cells transfected with miR-186 inhibitors with or without si-APAF1. Magnification, ×400. **P<0.01 vs. Inhibitor+NC. APAF1, apoptotic protease activating factor 1; LC3-B, light chain 3B; miR, microRNA; si, short interfering; NC, negative control.
Fig 2: miR-186 and APAF1 expression and their correlation in cSCC tissues. Reverse transcription-quantitative polymerase chain reaction was used to detect the expression of (A) miR-186 and (B) APAF1 in cSCC tissues and their corresponding controls. (C) Correlation between miR-186 expression and APAF1 expression. (D) Western blotting was performed to detect the protein expression of APAF1 in cSCC tissues and corresponding controls. ***P<0.001. miR, microRNA; cSCC, cutaneous squamous cell carcinoma; APAF1, apoptotic protease activating factor 1.
Fig 3: Analysis of apoptosis, drug resistance, and metastasis factors. Representative photomicrographs of immunohistochemistry of tumor fragments of mice receiving different treatments (A). Immunohistochemistry score by anti-FADD (B), anti-BCL-2 (C), anti-caspase-3 (D), relative messenger ribonucleic acids (mRNA) expression by RT-PCR for FADD and apoptotic protease activating factor 1 (APAF -1) (E), multidrug resistance protein 1 (MDR1) and survivin (F), and C-X-C chemokine receptor type 4 (CXCR4), and monocyte-derived chemokine (CCL22) (G). All treatment groups were compared to the negative control group (**** p < 0.0001). Comparison between OXA (5 mg/kg) and NPs 1 (5 mg/kg, ♦ p < 0.05) as well as between OXA (5 mg/kg) and NPs 2 (5 mg/kg, ♦♦ p < 0.01) was also performed (p < 0.0001 for both). Magnification: 40×.
Fig 4: Autophagy inhibitor treatment ameliorates exosome H/R injury-inhibiting effects. (A) EdU assay analysis of proliferation rates of H9c2 cells treated with 3-MA following H/R and exosome treatment (magnification, ×100). (B) Hoechst 33258 staining of apoptosis of H9c2 cells treated with 3-MA following H/R and exosome treatment (magnification, ×200). (C) Western blot analysis of apoptosis and autophagy-related protein expression in H9c2 cells treated with 3-MA after H/R and exosome treatment. GAPDH was used as the internal standard. **P<0.01 vs. control. H/R, hypoxia-reoxygenation; EdU, 5-ethynyl-2′-deoxyuridine; 3-MA, 3-Methyladenine; Apaf1, apoptotic protease activating factor-1; ATG13, autophagy-related protein 13; Bax, BCL2-associated X protein; H/R, hypoxia-reoxygenation.
Fig 5: Inhibition of rat myocardial damages by MSC-derived exosomes. (A) TTC staining and (B) H&E, Masson and TUNEL staining of the Sham, I/R and I/R+Exo groups (magnification, ×400). (C) Reverse transcription-quantitative PCR and (D) western blot analysis of Apaf1 and ATG13 mRNA and protein levels in the Sham, I/R and I/R+Exo groups. GAPDH was applied as the internal standard. **P<0.01 vs. Sham group; #P<0.05 vs. I/R group. MSC, mesenchymal stem cell; TTC, 2,3,5-triphenyltetrazolium chloride; H&E, hematoxylin and eosin; I/R, ischemia reperfusion; Exo, exosomes; Apaf1, apoptotic protease activating factor-1; ATG13, autophagy-related protein 13.
Supplier Page from Abcam for Anti-APAF1 antibody