Fig 2: HIF-1α activation strengthened the depressed effect of CNN1 in BC cells. (A) The protein levels of CNN1, HIF-1α, and PDK1 were tested by Western blotting in T24 and UMUC-3 cells. (B) T24 and UMUC-3 cell proliferation was detected by CCK8 assay. (C) T24 and UMUC-3 cell invasion was tested by transwell assay (bar = 50 µm). (D) T24 and UMUC-3 cell migration was detected by transwell assay (bar = 50 µm). (E) The protein levels of GLUT1, PKM2, and LDHA in T24 and UMUC-3 cells were tested by Western blotting. (F) The glucose consumption, (G) lactate production, and (H) ATP levels of T24 and UMUC-3 cells were tested by the corresponding kit. **p < 0.01, ***p < 0.001 vs. vector.

Fig 3: CNN1 overexpression suppresses BC cell growth in vivo. (A) Tumors were gathered from mice. (B) Tumor volume curve was analyzed. (C) Tumor weight was monitored. (D) Nude mouse tumor tissues were used for immunohistochemical experiments to detect the expression levels of Ki-67 and CNN1. **p < 0.01, ***p < 0.001 vs. vector.

Fig 4: The influences of CNN1 on BC cell properties and glycolysis. (A) qRT-PCR was utilized to determine the mRNA expression of CNN1 in T24 and UMUC-3 cells. (B) Western blotting was utilized to test the protein level of CNN1 in T24 and UMUC-3 cells. (C) The influence of CNN1 overexpression on T24 and UMUC-3 cell proliferation was tested by CCK8 assay. (D) The influences of CNN1 overexpression on T24 and UMUC-3 cell invasion were determined by transwell assay (bar = 50 µm). (E) The impacts of CNN1 overexpression on T24 and UMUC-3 cell migration were determined by transwell assay (bar = 50 µm). (F) The protein levels of GLUT1, PKM2, and LDHA in T24 and UMUC-3 cells were tested by Western blotting. (G) The glucose consumption, (H) lactate production, and (I) ATP levels of T24 and UMUC-3 cells were tested by the corresponding kit.*p < 0.05, **p < 0.01, ***p < 0.001 vs. vector.

Fig 5: CNN1 is lowly expressed in BC tissues and cells. (A) The qRT-PCR experiment was utilized to test the mRNA expression level of CNN1 in the cancerous tissues and adjacent normal tissues of BC patients. ***p < 0.001 vs. normal tissues. (B) Western blotting was utilized to test the protein level of CNN1 in the cancerous tissues and adjacent normal tissues of BC patients. ***p < 0.001 vs. normal tissues. (C) Kaplan–Meier analysis was utilized to inquiry the OS of CNN1 in BC patients, which were determined by Log-rank test. (D) CNN1 mRNA expression level in SV-HUC, EJ, T24, 5637, UMUC-3, and J82 cell lines. *p < 0.001, ***p < 0.001 vs. SV-HUC. (E) CNN1 protein expression level in SV-HUC, EJ, T24, 5637, UMUC-3, and J82 cell lines. *p < 0.05, **p < 0.01, ***p < 0.001 vs. SV-HUC.