Journal: Journal of Translational Medicine

Article Title: The miR-15b-5p/miR-379-3p-FOXO axis regulates cell cycle and apoptosis in scleral remodeling during experimental myopia

doi: 10.1186/s12967-024-05523-x

Figure Lengend Snippet: The primer sequence of the target genes

Article Snippet: Cyclin D1 (CCND1) , 1:1000 , Bioss, China.

Techniques: Sequencing

Journal: Journal of Translational Medicine

Article Title: The miR-15b-5p/miR-379-3p-FOXO axis regulates cell cycle and apoptosis in scleral remodeling during experimental myopia

doi: 10.1186/s12967-024-05523-x

Figure Lengend Snippet: List of Western blotting primary antibody

Article Snippet: Cyclin D1 (CCND1) , 1:1000 , Bioss, China.

Techniques: Western Blot

Journal: Journal of Translational Medicine

Article Title: The miR-15b-5p/miR-379-3p-FOXO axis regulates cell cycle and apoptosis in scleral remodeling during experimental myopia

doi: 10.1186/s12967-024-05523-x

Figure Lengend Snippet: List of Immunofluorescence staining primary antibody

Article Snippet: Cyclin D1 (CCND1) , 1:1000 , Bioss, China.

Techniques: Immunofluorescence, Staining

Journal: Journal of Translational Medicine

Article Title: The miR-15b-5p/miR-379-3p-FOXO axis regulates cell cycle and apoptosis in scleral remodeling during experimental myopia

doi: 10.1186/s12967-024-05523-x

Figure Lengend Snippet: Scleral cell cycle measurement and apoptotic analysis. A , B Measurement of CDK2 and CCND1 expression at the mRNA level in the sclera of the guinea pigs in NC and LIM groups (n = 4–6). C CDK2 and CCND1 immunoblots. D , E Measurement of CDK2 and CCND1 expression at protein level by Western blot in the sclera of the guinea pigs in NC and LIM groups (n = 6). F Measurement of CDK2 and CCND1 expression at protein level by immunofluorescence in the sclera of the guinea pigs in NC and LIM groups. (Western Blot cropped) (n = 6)

Article Snippet: Cyclin D1 (CCND1) , 1:1000 , Bioss, China.

Techniques: Expressing, Western Blot, Immunofluorescence

Journal: PeerJ

Article Title: Production of recombinant human epidermal growth factor fused with HaloTag protein and characterisation of its biological functions

doi: 10.7717/peerj.17806

Figure Lengend Snippet: (A) Validation of the differentially expressed genes by q-PCR with eight replicates. (B and C) Western blot analysis of the expression of proteins related to cell proliferation. (D and E) Cell immunofluorescence detection of the protein levels of c-Jun and CCND1. Three biological replicates were performed for Western blot assay. The p values of the comparison between control group and Halo-rhEGF group are all less than 0.05. Scale bar: 200 μm.

Article Snippet: The membrane was blocked with 5% (w/v) skimmed milk in TBST (20 mM Tris, 150 mM NaCl and 0.1% Tween in water, pH 7.5) and incubated with primary antibodies against ERK1/2, phos-ERK1/2, c-Jun, phos-c-Jun, Tubulin, Cyclin D1 (CCND1) at 4 °C for 24 h. The membranes were incubated with HRP-conjugated secondary antibodies at room temperature for 1 h and the labelled protein bands were visualized on Amersham Imager 600 (Amersham Biosciences, Piscataway, NJ, USA).

Techniques: Western Blot, Expressing, Immunofluorescence, Comparison, Control

Journal: Nagoya Journal of Medical Science

Article Title: Protective effect of Sasa veitchii extract against all-trans-retinoic acid-induced inhibition of proliferation of cultured human palate cells

doi: 10.18999/nagjms.86.2.223

Figure Lengend Snippet: Involvement with apoptosis and cell cycle by treatment with of SE against at RA in HEPM cells Fig. 3A: Apotracker staining (green) of HEPM cells after treatment with 100 μM at RA and 0.03% SE for 48 h. The nuclei were counterstained with Hoechst 33342 (blue). Scale bar, 50 μm. Fig. 3B: Immunoblotting of HEPM cells after treatment with 100 μM at RA and 0.03% SE for 48 h. β-actin was served as an internal control. *p < 0.05 and ***p < 0.001 versus at RA. at RA: all-trans -retinoic acid HEPM: human embryonic palatal mesenchymal SE: Sasa veitchii extract CCND1: cyclin D1

Article Snippet: Rabbit anti-cleaved caspase-3 polyclonal antibody (1:2,500 dilution; Cell Signaling Technology, Beverly, MA), mouse anti-cyclin D1 (CCND1) monoclonal antibody (1:1,000 dilution; Santa Cruz Biotechnology, Dallas, TX), mouse anti-CCNA (1:1,000 dilution; Santa Cruz Biotechnology), anti-mouse CCNB (1:1,000 dilution; Santa Cruz Biotechnology), anti-mouse CCNE (1:1,000 dilution; Santa Cruz Biotechnology), and anti-mouse β-actin monoclonal antibody (1:2,500 dilution; MBL, Aichi, Japan) were used as primary antibodies for immunoblotting.

Techniques: Staining, Western Blot, Control

Journal: Nagoya Journal of Medical Science

Article Title: Protective effect of Sasa veitchii extract against all-trans-retinoic acid-induced inhibition of proliferation of cultured human palate cells

doi: 10.18999/nagjms.86.2.223

Figure Lengend Snippet: Proposed mechanism of SE against at RA-induced cell proliferation inhibition ERBB2: Erb-B2 receptor tyrosine kinase 2 JADE1 : jade family PHD finger 1 CCND1: cyclin D1

Article Snippet: Rabbit anti-cleaved caspase-3 polyclonal antibody (1:2,500 dilution; Cell Signaling Technology, Beverly, MA), mouse anti-cyclin D1 (CCND1) monoclonal antibody (1:1,000 dilution; Santa Cruz Biotechnology, Dallas, TX), mouse anti-CCNA (1:1,000 dilution; Santa Cruz Biotechnology), anti-mouse CCNB (1:1,000 dilution; Santa Cruz Biotechnology), anti-mouse CCNE (1:1,000 dilution; Santa Cruz Biotechnology), and anti-mouse β-actin monoclonal antibody (1:2,500 dilution; MBL, Aichi, Japan) were used as primary antibodies for immunoblotting.

Techniques: Inhibition

Journal: APL Bioengineering

Article Title: Survivin as a mediator of stiffness-induced cell cycle progression and proliferation of vascular smooth muscle cells

doi: 10.1063/5.0150532

Figure Lengend Snippet: Survivin is required for stiffness-mediated cell cycle progression and proliferation. (a)–(g) hVSMCs were transfected with control siRNA or siRNAs to survivin [#1 and #2], synchronized to G 0 via serum starvation, and plated on soft or stiff hydrogels with 10% FBS for 24 h. Total cell lysates were analyzed by RT-qPCR (a)–(c) or immunoblotting (d)–(g) to determine mRNA and protein levels of survivin (a) and (e), cyclin D1 [CCND1; (b) and (f)], and cyclin A [CCNA; (c) and (g)]. Expression levels were normalized to those in hVSMCs treated with control siRNA on stiff hydrogels. n = 4 (a), n = 3 (b), n = 3 (c), n = 5 (e), n = 3 (f), and n = 5 (g), independent experiments. S-phase entry and cell proliferation with survivin knockdown (h)–(i) and overexpression (j)–(k) were assessed by EdU incorporation (h) and (j) and cell counting (i) and (k), respectively, and normalized to values from hVSMCs treated with control siRNA on stiff hydrogels (h)–(i) or infected with GFP adenovirus (Adv) on soft hydrogels ( j)–(k). n = 5 (h), n = 3 (i), n = 4 (j), and n = 4 (k), independent experiments. Data are means + SEMs. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: The following survivin ( BIRC5 ), FAK ( PTK2 ), and cyclin D1 ( CCND1 ) siRNAs were obtained from Ambion: survivin siRNA #1 (ID no. 121294), 5′-CCACUUCCAGGGUUUAUUCtt-3′; survivin siRNA #2 (ID no. 121295), 5′-GCCAUUCUAAGUCAUUGGGtt-3′; FAK siRNA #1 (ID no. 157448), 5′-CCUAGCAGACUUUAACCAAtt-3′; FAK siRNA #2 (ID no. 61352), 5′-GGCAUGGAGAUGCUACUGAtt-3′; and cyclin D1 siRNA (ID no. 42828), 5′-GGAGAACAAACAGAUCAUCtt-3′.

Techniques: Transfection, Quantitative RT-PCR, Western Blot, Expressing, Over Expression, Cell Counting, Infection

Journal: APL Bioengineering

Article Title: Survivin as a mediator of stiffness-induced cell cycle progression and proliferation of vascular smooth muscle cells

doi: 10.1063/5.0150532

Figure Lengend Snippet: Survivin is an upstream regulator of stiffness-induced cell cycle progression. (a) and (b) hVSMCs were transfected with control siRNA or siRNAs to cyclin D1, synchronized to G 0 via serum starvation, and plated on stiff hydrogels with 10% FBS for 24 h. Total cell lysates were analyzed by RT-qPCR to determine mRNA levels of cyclin D1 (CCND1) (a) and survivin (b). Expression levels were normalized to those in hVSMCs treated with control siRNA on stiff hydrogels. n = 3, independent experiments. (c) G 0 -synchronized hVSMCs were treated with either 5 or 30 μ M mitomycin C (MMC) and then cultured on soft or stiff hydrogels for 24 h. Survivin induction was analyzed by immunoblotting. Survivin protein levels were normalized to those in hVSMCs treated with DMSO on stiff hydrogels (d). n = 4–8, independent experiments.

Article Snippet: The following survivin ( BIRC5 ), FAK ( PTK2 ), and cyclin D1 ( CCND1 ) siRNAs were obtained from Ambion: survivin siRNA #1 (ID no. 121294), 5′-CCACUUCCAGGGUUUAUUCtt-3′; survivin siRNA #2 (ID no. 121295), 5′-GCCAUUCUAAGUCAUUGGGtt-3′; FAK siRNA #1 (ID no. 157448), 5′-CCUAGCAGACUUUAACCAAtt-3′; FAK siRNA #2 (ID no. 61352), 5′-GGCAUGGAGAUGCUACUGAtt-3′; and cyclin D1 siRNA (ID no. 42828), 5′-GGAGAACAAACAGAUCAUCtt-3′.

Techniques: Transfection, Quantitative RT-PCR, Expressing, Cell Culture, Western Blot