Cyclin E1 Antibody - #AF0144

Figure 3 Reduction in ATP production hinders cell proliferation and contributes to G1/S arrest in GBM. (A-B) The relative viability of TBD0220 (A) and U87MG (B) cells was measured using a CCK-8 kit (n = 3). (C-D) Colony formation assay to detect GBM cell growth (D), and the quantification of colony numbers (C) (n = 3). (E) Cell cycle analysis using flow cytometry after incubation with different treatments like EPIC (20 µM), AA (25 µM), or EPIC (20 µM) + AA (25 µM) for 48 h, and the results are plotted as a histogram (n = 3) (F) Representative western blotting showing the expression of p21 and Rb and their downstream targets. The results were normalized to Tubulin with the control group as 1. Protein expression was quantified by ImageJ. (G) Representative confocal images of CDK6 after the treatment with AA (25 µM), EPIC (20 µM), or EPIC (20 µM) + AA (25 µM) for 48 h (n = 6). (H) The quantitative analysis of fluorescence images of CDK6 (n = 6). All data are shown as the mean values ± SD, and p values are based on one-way or two-way ANOVA. ****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05. Scale bar = 20 μm.

Fig. 2 Functional effects of lncRNA RP11-197K6.1 knockdown in CRC cells. A: Confirmation of lncRNA RP11-197K6.1 knockdown efficiency in HCT116 cells by RT-qPCR. B: Reduction in the proliferation of HCT116 and SW480 cells post-knockdown, as determined by proliferation assays. C: Increase in the apoptosis of HCT116 and SW480 cells following lncRNA RP11-197K6.1 knockdown, as measured by flow cytometry. D & E: Decrease in the migration and invasion of HCT116 and SW480 cells post-knockdown, as assessed by wound healing (scale = 200 μm) and Transwell assays (scale = 50 μm), respectively. 2 F: Changes in the levels of proteins related to the proliferation, apoptosis, and migration of HCT116 and SW480 cells after lncRNA RP11-197K6.1 knockdown, as analyzed by western blotting assay (**P 

Fig. 5. |SiNPs affected the protein expressions of cell cycle regulators. (A) The protein expressions of cyclin A1, CDK2, cyclin E1, CDK1, and cyclin B1 were significantly decreased in the testis after exposure to SiNPs.

Figure 4 |The loss of RECQL4 induces cell cycle arrest and cellular senescence. . (D) The protein levels of c-myc, p21, cyclin D, CDK6, cyclin E, Bax, and Bcl-2 were determined by Western blot in stable Tet-on inducible RECQL4 knockdown cell lines (KYSE30 and TE-1 cells) (+Dox) and controls (–Dox). Experiments were independently repeated 3 times. All data indicate the mean ±SD. *P < 0.05; **P < 0.01; ***P < 0.001.

Figure 4 The loss of RECQL4 induces cell cycle arrest and cellular senescence. (A) Depletion of RECQL4 by siRNA. RECQL4 protein levels were measured by Western blot. KYSE30 and TE-1 cells were transfected with siRNA duplexes (200 nM) specific to RECQL4 or negative oligo in serum-free medium for 4 h, then replaced with complete medium for 24 h. Whole cell extracts were collected for Western blot analysis using RECQL4 antibodies. (B) Cell cycle distributions in RECQL4 knockdown cell lines (KYSE30 and TE-1 cells) and controls were determined by flow cytometry. (C) Cellular senescence was examined by SA-β-gal staining. Microscopic magnification (×200), Scale bar: 50 μm. (D) The protein levels of c-myc, p21, cyclin D, CDK6, cyclin E, Bax, and Bcl-2 were determined by Western blot in stable Tet-on inducible RECQL4 knockdown cell lines (KYSE30 and TE-1 cells) (+Dox) and controls (–Dox). Experiments were independently repeated 3 times. All data indicate the mean ± SD. *P < 0.05; **P < 0.01; ***P < 0.001.

Figure 4 OTUB1 promotes and regulates the expression of Cyclin E1. (A) The relationship between OTUB1 and Cyclin E1 was analyzed via gene MANIA database. (B) The protein expression of Cyclin E1 in BPH, ADPC, and CRPC was detected by IHC staining. (C) Western blotting detecting the expression of Cyclin E1 in PC3 and C4-2 cells transfected with otub1 overexpression and otub1 c91s. (D) Western blotting detecting the expression of Cyclin E1 in PC3 and C4-2 cells transfected with OTUB1 siRNA. (E) Western blotting detecting the expression of Cyclin E1 in PC3 and C4-2 cells transfected with an increasing gradient OTUB1.

Figure 7 OTUB1 promotes the growth of PC3 cell via increasing Cyclin E1 in vivo. (A,B) PC3 cells transfected with OTUB1 overexpression and otub1 c91s were transplanted subcutaneously in nude mice. The effect of otub1 overexpression, otub1 c91s, and RO-3306 on the growth of PCa. (C) The tumor volume was measured with a caliper. (D) The tumor volumes were measured daily when the diameter grew into 2 mm. (E) IHC staining detecting the expression of OTUB1, ki-67, and Cyclin E1 in these tumors from nude mice. ****P < 0.0001.