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Compound 053 enhances the therapeutic effect of GEM by inhibiting autophagy in the MDA-MB-231 xenograft model. (A) Mouse treatment plan. (B) The tumor size was measured with a caliper every 3 days and the volume was calculated (n=5). (C) Tumor morphology of mouse removed at 21 days of treatment. (D) The tumors were removed and weighed at 21 days of treatment. Data are presented as mean ± SEM (n=5). Statistical significance was determined by one-way ANOVA followed by Dunnett's multiple comparisons test to compare all experimental groups against a single control group. *** P<0.001 vs. Control. (E) Protein levels of Ki67, LC3B, p62, PD-L1 and <t>PD-L2</t> were detected using immunohistochemical staining in tumor tissues of mice after 21 days of treatment. Nuclei are localized in blue and target proteins are localized in brown. Scale bar, 50 μ m. (F) Body weight was measured every 3 days. (G) The TUNEL assay was used to evaluate DNA fragmentation in mouse tumor tissues after 21 days of treatment. Blue fluorescence indicates nuclear staining (DAPI), while green fluorescence specifically labels DNA breaks associated with apoptosis. Scale bar, 100 μ m. (H) H&E staining of tumor tissues and major organs (heart, liver, spleen, lungs and kidneys) of mice after 21 days of treatment. Nuclei are blue-purple and cytoplasm is red. Scale bar, 100 μ m. 053, FZU-0045-053; GEM, gemcitabine; LC3B, microtubule-associated protein 1A/1B-light chain 3B; p62/SQSTM1, sequestosome 1; PD-L1/2, programmed death-ligand 1/2.
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Compound 053 enhances the therapeutic effect of GEM by inhibiting autophagy in the MDA-MB-231 xenograft model. (A) Mouse treatment plan. (B) The tumor size was measured with a caliper every 3 days and the volume was calculated (n=5). (C) Tumor morphology of mouse removed at 21 days of treatment. (D) The tumors were removed and weighed at 21 days of treatment. Data are presented as mean ± SEM (n=5). Statistical significance was determined by one-way ANOVA followed by Dunnett's multiple comparisons test to compare all experimental groups against a single control group. *** P<0.001 vs. Control. (E) Protein levels of Ki67, LC3B, p62, PD-L1 and <t>PD-L2</t> were detected using immunohistochemical staining in tumor tissues of mice after 21 days of treatment. Nuclei are localized in blue and target proteins are localized in brown. Scale bar, 50 μ m. (F) Body weight was measured every 3 days. (G) The TUNEL assay was used to evaluate DNA fragmentation in mouse tumor tissues after 21 days of treatment. Blue fluorescence indicates nuclear staining (DAPI), while green fluorescence specifically labels DNA breaks associated with apoptosis. Scale bar, 100 μ m. (H) H&E staining of tumor tissues and major organs (heart, liver, spleen, lungs and kidneys) of mice after 21 days of treatment. Nuclei are blue-purple and cytoplasm is red. Scale bar, 100 μ m. 053, FZU-0045-053; GEM, gemcitabine; LC3B, microtubule-associated protein 1A/1B-light chain 3B; p62/SQSTM1, sequestosome 1; PD-L1/2, programmed death-ligand 1/2.
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Compound 053 enhances the therapeutic effect of GEM by inhibiting autophagy in the MDA-MB-231 xenograft model. (A) Mouse treatment plan. (B) The tumor size was measured with a caliper every 3 days and the volume was calculated (n=5). (C) Tumor morphology of mouse removed at 21 days of treatment. (D) The tumors were removed and weighed at 21 days of treatment. Data are presented as mean ± SEM (n=5). Statistical significance was determined by one-way ANOVA followed by Dunnett's multiple comparisons test to compare all experimental groups against a single control group. *** P<0.001 vs. Control. (E) Protein levels of Ki67, LC3B, p62, PD-L1 and <t>PD-L2</t> were detected using immunohistochemical staining in tumor tissues of mice after 21 days of treatment. Nuclei are localized in blue and target proteins are localized in brown. Scale bar, 50 μ m. (F) Body weight was measured every 3 days. (G) The TUNEL assay was used to evaluate DNA fragmentation in mouse tumor tissues after 21 days of treatment. Blue fluorescence indicates nuclear staining (DAPI), while green fluorescence specifically labels DNA breaks associated with apoptosis. Scale bar, 100 μ m. (H) H&E staining of tumor tissues and major organs (heart, liver, spleen, lungs and kidneys) of mice after 21 days of treatment. Nuclei are blue-purple and cytoplasm is red. Scale bar, 100 μ m. 053, FZU-0045-053; GEM, gemcitabine; LC3B, microtubule-associated protein 1A/1B-light chain 3B; p62/SQSTM1, sequestosome 1; PD-L1/2, programmed death-ligand 1/2.
Pd L2 Primary Antibodies, supplied by Proteintech, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Compound 053 enhances the therapeutic effect of GEM by inhibiting autophagy in the MDA-MB-231 xenograft model. (A) Mouse treatment plan. (B) The tumor size was measured with a caliper every 3 days and the volume was calculated (n=5). (C) Tumor morphology of mouse removed at 21 days of treatment. (D) The tumors were removed and weighed at 21 days of treatment. Data are presented as mean ± SEM (n=5). Statistical significance was determined by one-way ANOVA followed by Dunnett's multiple comparisons test to compare all experimental groups against a single control group. *** P<0.001 vs. Control. (E) Protein levels of Ki67, LC3B, p62, PD-L1 and <t>PD-L2</t> were detected using immunohistochemical staining in tumor tissues of mice after 21 days of treatment. Nuclei are localized in blue and target proteins are localized in brown. Scale bar, 50 μ m. (F) Body weight was measured every 3 days. (G) The TUNEL assay was used to evaluate DNA fragmentation in mouse tumor tissues after 21 days of treatment. Blue fluorescence indicates nuclear staining (DAPI), while green fluorescence specifically labels DNA breaks associated with apoptosis. Scale bar, 100 μ m. (H) H&E staining of tumor tissues and major organs (heart, liver, spleen, lungs and kidneys) of mice after 21 days of treatment. Nuclei are blue-purple and cytoplasm is red. Scale bar, 100 μ m. 053, FZU-0045-053; GEM, gemcitabine; LC3B, microtubule-associated protein 1A/1B-light chain 3B; p62/SQSTM1, sequestosome 1; PD-L1/2, programmed death-ligand 1/2.
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Compound 053 enhances the therapeutic effect of GEM by inhibiting autophagy in the MDA-MB-231 xenograft model. (A) Mouse treatment plan. (B) The tumor size was measured with a caliper every 3 days and the volume was calculated (n=5). (C) Tumor morphology of mouse removed at 21 days of treatment. (D) The tumors were removed and weighed at 21 days of treatment. Data are presented as mean ± SEM (n=5). Statistical significance was determined by one-way ANOVA followed by Dunnett's multiple comparisons test to compare all experimental groups against a single control group. *** P<0.001 vs. Control. (E) Protein levels of Ki67, LC3B, p62, PD-L1 and <t>PD-L2</t> were detected using immunohistochemical staining in tumor tissues of mice after 21 days of treatment. Nuclei are localized in blue and target proteins are localized in brown. Scale bar, 50 μ m. (F) Body weight was measured every 3 days. (G) The TUNEL assay was used to evaluate DNA fragmentation in mouse tumor tissues after 21 days of treatment. Blue fluorescence indicates nuclear staining (DAPI), while green fluorescence specifically labels DNA breaks associated with apoptosis. Scale bar, 100 μ m. (H) H&E staining of tumor tissues and major organs (heart, liver, spleen, lungs and kidneys) of mice after 21 days of treatment. Nuclei are blue-purple and cytoplasm is red. Scale bar, 100 μ m. 053, FZU-0045-053; GEM, gemcitabine; LC3B, microtubule-associated protein 1A/1B-light chain 3B; p62/SQSTM1, sequestosome 1; PD-L1/2, programmed death-ligand 1/2.
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Compound 053 enhances the therapeutic effect of GEM by inhibiting autophagy in the MDA-MB-231 xenograft model. (A) Mouse treatment plan. (B) The tumor size was measured with a caliper every 3 days and the volume was calculated (n=5). (C) Tumor morphology of mouse removed at 21 days of treatment. (D) The tumors were removed and weighed at 21 days of treatment. Data are presented as mean ± SEM (n=5). Statistical significance was determined by one-way ANOVA followed by Dunnett's multiple comparisons test to compare all experimental groups against a single control group. *** P<0.001 vs. Control. (E) Protein levels of Ki67, LC3B, p62, PD-L1 and <t>PD-L2</t> were detected using immunohistochemical staining in tumor tissues of mice after 21 days of treatment. Nuclei are localized in blue and target proteins are localized in brown. Scale bar, 50 μ m. (F) Body weight was measured every 3 days. (G) The TUNEL assay was used to evaluate DNA fragmentation in mouse tumor tissues after 21 days of treatment. Blue fluorescence indicates nuclear staining (DAPI), while green fluorescence specifically labels DNA breaks associated with apoptosis. Scale bar, 100 μ m. (H) H&E staining of tumor tissues and major organs (heart, liver, spleen, lungs and kidneys) of mice after 21 days of treatment. Nuclei are blue-purple and cytoplasm is red. Scale bar, 100 μ m. 053, FZU-0045-053; GEM, gemcitabine; LC3B, microtubule-associated protein 1A/1B-light chain 3B; p62/SQSTM1, sequestosome 1; PD-L1/2, programmed death-ligand 1/2.
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Compound 053 enhances the therapeutic effect of GEM by inhibiting autophagy in the MDA-MB-231 xenograft model. (A) Mouse treatment plan. (B) The tumor size was measured with a caliper every 3 days and the volume was calculated (n=5). (C) Tumor morphology of mouse removed at 21 days of treatment. (D) The tumors were removed and weighed at 21 days of treatment. Data are presented as mean ± SEM (n=5). Statistical significance was determined by one-way ANOVA followed by Dunnett's multiple comparisons test to compare all experimental groups against a single control group. *** P<0.001 vs. Control. (E) Protein levels of Ki67, LC3B, p62, PD-L1 and <t>PD-L2</t> were detected using immunohistochemical staining in tumor tissues of mice after 21 days of treatment. Nuclei are localized in blue and target proteins are localized in brown. Scale bar, 50 μ m. (F) Body weight was measured every 3 days. (G) The TUNEL assay was used to evaluate DNA fragmentation in mouse tumor tissues after 21 days of treatment. Blue fluorescence indicates nuclear staining (DAPI), while green fluorescence specifically labels DNA breaks associated with apoptosis. Scale bar, 100 μ m. (H) H&E staining of tumor tissues and major organs (heart, liver, spleen, lungs and kidneys) of mice after 21 days of treatment. Nuclei are blue-purple and cytoplasm is red. Scale bar, 100 μ m. 053, FZU-0045-053; GEM, gemcitabine; LC3B, microtubule-associated protein 1A/1B-light chain 3B; p62/SQSTM1, sequestosome 1; PD-L1/2, programmed death-ligand 1/2.
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Compound 053 enhances the therapeutic effect of GEM by inhibiting autophagy in the MDA-MB-231 xenograft model. (A) Mouse treatment plan. (B) The tumor size was measured with a caliper every 3 days and the volume was calculated (n=5). (C) Tumor morphology of mouse removed at 21 days of treatment. (D) The tumors were removed and weighed at 21 days of treatment. Data are presented as mean ± SEM (n=5). Statistical significance was determined by one-way ANOVA followed by Dunnett's multiple comparisons test to compare all experimental groups against a single control group. *** P<0.001 vs. Control. (E) Protein levels of Ki67, LC3B, p62, PD-L1 and PD-L2 were detected using immunohistochemical staining in tumor tissues of mice after 21 days of treatment. Nuclei are localized in blue and target proteins are localized in brown. Scale bar, 50 μ m. (F) Body weight was measured every 3 days. (G) The TUNEL assay was used to evaluate DNA fragmentation in mouse tumor tissues after 21 days of treatment. Blue fluorescence indicates nuclear staining (DAPI), while green fluorescence specifically labels DNA breaks associated with apoptosis. Scale bar, 100 μ m. (H) H&E staining of tumor tissues and major organs (heart, liver, spleen, lungs and kidneys) of mice after 21 days of treatment. Nuclei are blue-purple and cytoplasm is red. Scale bar, 100 μ m. 053, FZU-0045-053; GEM, gemcitabine; LC3B, microtubule-associated protein 1A/1B-light chain 3B; p62/SQSTM1, sequestosome 1; PD-L1/2, programmed death-ligand 1/2.

Journal: International Journal of Oncology

Article Title: Discovery of the late autophagy inhibitor FZU-0045-053 and its anti-breast cancer and immunomodulatory effects

doi: 10.3892/ijo.2025.5823

Figure Lengend Snippet: Compound 053 enhances the therapeutic effect of GEM by inhibiting autophagy in the MDA-MB-231 xenograft model. (A) Mouse treatment plan. (B) The tumor size was measured with a caliper every 3 days and the volume was calculated (n=5). (C) Tumor morphology of mouse removed at 21 days of treatment. (D) The tumors were removed and weighed at 21 days of treatment. Data are presented as mean ± SEM (n=5). Statistical significance was determined by one-way ANOVA followed by Dunnett's multiple comparisons test to compare all experimental groups against a single control group. *** P<0.001 vs. Control. (E) Protein levels of Ki67, LC3B, p62, PD-L1 and PD-L2 were detected using immunohistochemical staining in tumor tissues of mice after 21 days of treatment. Nuclei are localized in blue and target proteins are localized in brown. Scale bar, 50 μ m. (F) Body weight was measured every 3 days. (G) The TUNEL assay was used to evaluate DNA fragmentation in mouse tumor tissues after 21 days of treatment. Blue fluorescence indicates nuclear staining (DAPI), while green fluorescence specifically labels DNA breaks associated with apoptosis. Scale bar, 100 μ m. (H) H&E staining of tumor tissues and major organs (heart, liver, spleen, lungs and kidneys) of mice after 21 days of treatment. Nuclei are blue-purple and cytoplasm is red. Scale bar, 100 μ m. 053, FZU-0045-053; GEM, gemcitabine; LC3B, microtubule-associated protein 1A/1B-light chain 3B; p62/SQSTM1, sequestosome 1; PD-L1/2, programmed death-ligand 1/2.

Article Snippet: The sections were then blocked with 5% BSA for 1 h at room temperature and subsequently incubated overnight at 4°C with the following primary antibodies: Anti-Ki67 antibody (cat. no. GB111499-100; 1:1,000; Wuhan Servicebio Technology Co., Ltd), anti-LC3 antibody (cat. no. 81004-1-RR; 1:1,000; Proteintech Group, Inc), anti-p62 antibody (cat. no. GB11239-1-100; 1:1,000; Wuhan Servicebio Technology Co., Ltd), anti-PD-L1 antibody (cat. no. GB150059-100; 1:1,000; Wuhan Servicebio Technology Co., Ltd), anti-PD-L2 antibody (cat. no. 27406-1-AP; 1:4,000; Proteintech Group, Inc), anti-CD3 antibody (cat. no. GB11014-100; 1:1,000; Wuhan Servicebio Technology Co., Ltd), anti-CD4 antibody (cat. no. GB11064-100; 1:1,000; Wuhan Servicebio Technology Co., Ltd), anti-CD8 antibody (cat. no. GB11068; 1:1,000; Wuhan Servicebio Technology Co., Ltd), anti-4-1BB/CD137 antibody (cat. no. A2025; 1:200; ABclonal Biotech Co., Ltd), anti-CD134/OX40 antibody (cat. no. 32621-1-AP; 1:500; Proteintech Group, Inc), anti-PD1 antibody (cat. no. GB153744-100; 1:400; Wuhan Servicebio Technology Co., Ltd) and anti-TIM3 antibody (cat. no. 11872-1-AP; 1:1,000; Proteintech Group, Inc).

Techniques: Control, Immunohistochemical staining, Staining, TUNEL Assay, Fluorescence