rabbit polyclonal anti cdk5  (Bioss)

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Effect of Sirtuin-1 on Synaptic Plasticity in Nucleus Accumbens in a Rat Model of Heroin Addiction

doi: 10.12659/MSM.910550

Figure Lengend Snippet: Cells positive for SIRT1, Cdk5, NF-κB, FOXO1, Cbp, and Syn in the NAc region (×400) and their optical density values ( A ) Saline control (SC) group; ( B ) Heroin-addicted (HA) group; ( C ) AAV-rSirt1 group; ( D ) AAV-rSirt1 shRNA group. # P <0.05 compared with the SC group; * P <0.05 compared with the HA group.

Article Snippet: The reagents and analysis systems used were as follows: heroin (purity 92.09%, provided by the Guizhou Public Security Bureau), pentobarbital sodium (Sigma, USA), naloxone hydrochloride (Chongqing YaoPharma, China), mouse monoclonal anti-SIRT1 (Abcam, USA), rabbit polyclonal anti-Cdk5, rabbit polyclonal anti-NF-κB (Bioss, China), rabbit monoclonal anti-FOXO1 (Abcam, USA), rabbit polyclonal anti-Cbp (Bioss, China), rabbit monoclonal anti-PSD95 (Abcam, USA), rabbit monoclonal anti-Syn (Abcam, USA), mouse monoclonal anti-beta-actin (Abcam, USA), SuperScript III RT reverse transcription kit (Invitrogen, USA) Sybr qPCR mix (Invitrogen, USA), Plasmid Mini kit (TransGen Biotech, Beijing, China), gel extraction kit (Omega), Trans2K Plus II DNA Marker (TransGen, Beijing, China), DNA primers (Invitrogen, USA), DNA sequencing (Invitrogen, USA), restriction endonuclease EcoRI (NEB, USA), T4 DNA Ligase (NEB, USA), Plasmid Maxi Kit (Qiagen, Germany), CPT High-efficiency Transfection Kit (Virotherapy Technologies, Wuhan, China).

Techniques: shRNA

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Effect of Sirtuin-1 on Synaptic Plasticity in Nucleus Accumbens in a Rat Model of Heroin Addiction

doi: 10.12659/MSM.910550

Figure Lengend Snippet: Detection of SIRT1, Cdk5, FOXO1, NF-κB, Syn, and PSD95 protein levels in each group by immunoblotting. # P <0.05 compared with the saline control group; * P <0.05 compared with the HA group .

Article Snippet: The reagents and analysis systems used were as follows: heroin (purity 92.09%, provided by the Guizhou Public Security Bureau), pentobarbital sodium (Sigma, USA), naloxone hydrochloride (Chongqing YaoPharma, China), mouse monoclonal anti-SIRT1 (Abcam, USA), rabbit polyclonal anti-Cdk5, rabbit polyclonal anti-NF-κB (Bioss, China), rabbit monoclonal anti-FOXO1 (Abcam, USA), rabbit polyclonal anti-Cbp (Bioss, China), rabbit monoclonal anti-PSD95 (Abcam, USA), rabbit monoclonal anti-Syn (Abcam, USA), mouse monoclonal anti-beta-actin (Abcam, USA), SuperScript III RT reverse transcription kit (Invitrogen, USA) Sybr qPCR mix (Invitrogen, USA), Plasmid Mini kit (TransGen Biotech, Beijing, China), gel extraction kit (Omega), Trans2K Plus II DNA Marker (TransGen, Beijing, China), DNA primers (Invitrogen, USA), DNA sequencing (Invitrogen, USA), restriction endonuclease EcoRI (NEB, USA), T4 DNA Ligase (NEB, USA), Plasmid Maxi Kit (Qiagen, Germany), CPT High-efficiency Transfection Kit (Virotherapy Technologies, Wuhan, China).

Techniques: Western Blot

Journal: Frontiers in Aging Neuroscience

Article Title: The γ-Adducin 1–357 fragment promotes tau pathology

doi: 10.3389/fnagi.2023.1241750

Figure Lengend Snippet: Antibody information.

Article Snippet: Rabbit Polyclonal anti-CDK5 , Proteintech , 10,430-1-AP , 1:1000 for Western blot.

Techniques: Western Blot, Immunofluorescence

Journal: Molecular Vision

Article Title: Cyclin-dependent kinase 5 promotes the stability of corneal epithelial cell junctions

doi:

Figure Lengend Snippet: Co-localization of E-cadherin and cyclin dependent kinase5 (Cdk5) in the cell–cell borders. Human corneal limbal epithelial (HCLE) cells were grown on chamber slides, fixed, and immunostained stained with pY15 (green; A ) and E-cadherin (red; B ). Cyclin-dependent kinase 5 (Cdk5) and E-cadherin were localized to the cell–cell boundaries, and the overlay image demonstrates the colocalization ( C ) of E-cadherin and phosphorylated (pY15) CDK5. Confluent cultures of HCLE showing CDK5 border localization ( D ) confluent HCLE cultures when treated with olomoucine show a shift in the localization of pY15 CDK5 from the cell borders to the interior ( E ). E-cadherin and CDK5 form a part of the same protein complex ( F , G ) E-cadherin coimmuneprecipitates with CDK5 (38 kDa; F). Affinity chromatography for HCLE cell lysates were pulled down for glutathione-S-transferase-cyclin-dependent kinase 5 (GST-CDK5) and that glutathione beads bound to CDK5 formed a complex with E-cadherin ( G ). ( A , B , C ) 100X images and ( D , E ) 40X images.

Article Snippet: The following antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA): anti-Cdk5 mouse monoclonal (DC-17; sc-249), anti-Cdk5 rabbit polyclonal (C-8, sc-173), anti-pY15-Cdk5 (sc-12918), and anti-p35 rabbit polyclonal (C-19).

Techniques: Staining, Affinity Chromatography

Journal: Molecular Vision

Article Title: Cyclin-dependent kinase 5 promotes the stability of corneal epithelial cell junctions

doi:

Figure Lengend Snippet: Human corneal limbal epithelial cell line suppressed for cyclin-dependent kinase 5 expression. Immunofluorescence image showing staining for cyclin-dependent kinase 5 (CDK5) (green; A ) in human corneal limbal epithelial (HCLE) cells and note the absence of Cdk5 in the ShCDK5 cells ( C ). Nuclear counterstaining 4', 6-diamidino-2-phenylindole (blue) for images in A and C ( B , D ). Lysates from these cells showing significant reduction in the expression levels of CDK5 in the plentiviral transduced HCLE (ShCDK5) cells ( E ). 100X images.

Article Snippet: The following antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA): anti-Cdk5 mouse monoclonal (DC-17; sc-249), anti-Cdk5 rabbit polyclonal (C-8, sc-173), anti-pY15-Cdk5 (sc-12918), and anti-p35 rabbit polyclonal (C-19).

Techniques: Expressing, Immunofluorescence, Staining

Journal: Molecular Vision

Article Title: Cyclin-dependent kinase 5 promotes the stability of corneal epithelial cell junctions

doi:

Figure Lengend Snippet: Cell-cell junction formation in the ShCDK5 human corneal limbal epithelial cells. Human corneal limbal epithelial (HCLE) cells ( A , B ), in the absence of cyclin-dependent kinase 5 (CDK5) with olomoucine ( C , D ) or ShCDK5 ( E , F ) and lens epithelial cells (NN1003; G , H ) were treated with olomoucine ( I, J ). The cells were allowed to form cell–cell adhesions in the presence or absence of calcium for 1 h at 37 °C, and particle aggregates were cytospin on to glass slides to measure the size of the aggregates. Calcium-dependent E-cadherin junctions were formed in the presence of CDK5 ( B , K ) in the HCLE cells in contrast to N-cadherin expressing lens epithelial cells that form larger aggregates in the absence of CDK5 ( J , K ). ( A – J ) 20X images. E-cadherin and N-cadherin induction during cell–cell junction formation in the MDA-MB-231 cells ( L ). Green fluorescent protein (GFP)-E-cadherin and GFP-N-cadherin transfected cells were allowed to form cell–cell aggregates in the absence of calcium (lane 1), presence (lane 2) and with olomoucine (lane 3). Cells containing E-cadherin form more cell–cell aggregates (group 3) while the N-cadherin (group 4) aggregates increases in the presence of olomoucine. N-cadherin induction in the ShCDK5 and olomoucine HCLE ( M ) CDK5 inhibition leading to degradation of E-cadherin leads to induction of N-cadherin. P120 coimmuneprecipitates with N-cadherin to stabilize the junctional complex in the ShCDK5 and olomoucine-treated HCLE. IP=immunoprecipitation; WCl=whole cell lysate.

Article Snippet: The following antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA): anti-Cdk5 mouse monoclonal (DC-17; sc-249), anti-Cdk5 rabbit polyclonal (C-8, sc-173), anti-pY15-Cdk5 (sc-12918), and anti-p35 rabbit polyclonal (C-19).

Techniques: Expressing, Transfection, Inhibition, Immunoprecipitation

Journal: Molecular Vision

Article Title: Cyclin-dependent kinase 5 promotes the stability of corneal epithelial cell junctions

doi:

Figure Lengend Snippet: Total internal reflection fluorescence analysis for the E-cadherin internalization pathway in the human corneal limbal epithelial and ShCDK5 cells. Green fluorescent protein (GFP)-E-cadherin transfected ( A – B ) epithelial cells with and without olomoucine and ShCDK5 cultures were analyzed for E-cadherin particle movement for distance and tortuosity from the cell–cell borders at a constant time ( A ). The tortuosity or particle tracking of the E-cadherin-containing vesicles was recorded. The E-cadherin vesicles moved in long paths (arrows) from the cell borders to the interior, and such long paths were significantly greater in the ShCDK5 suggestive of internalization than the control human corneal limbal epithelial (HCLE) cells. Total internal reflection fluorescence (TIRF) analysis for the p120 ( C – H ) shows internalization pathway in the HCLE and ShCDK5. A GFP-p120 clone was transfected into the HCLE and ShCDK5 cells, and the vesicle tracking (arrows) for the tortuosity was measured. P120 vesicles remained in the cell–cell borders, and no long paths were internalized. In the HCLE and ShCDK5 cells, the p120 tortuosity paths were restricted within the cell–cell borders. Panel inserts in the HCLE cells transfected with p120 ( C ) and ShCdk5 transfected with p120 ( E ) are enlarged (zoom to a factor of 2) and shown in D and F , respectively, along with the tortuosity path for each junctional vesicle analyzed. The distance and tortuosity of paths by the E-cadherin- and p120-containing particles (n>100) were tracked at a constant time for all the particles analyzed using TIRF microscopy are described in . The E-cadherin-containing particles in the ShCdk5 cells moved fast, and these long, straight paths were internalized ( B ). The p120-containing particles in the ShCdk5 cells moved slowly and stayed near the cell–cell boundary ( C–F ). In the absence of CDK5, the E-cadherin and p120 junctional complex dissociate, promoting internalization and endocytosis of E-cadherin. Cdk5 stabilizes the junction by preventing E-cadherin-containing vesicles from being endocytosed. 100X images.

Article Snippet: The following antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA): anti-Cdk5 mouse monoclonal (DC-17; sc-249), anti-Cdk5 rabbit polyclonal (C-8, sc-173), anti-pY15-Cdk5 (sc-12918), and anti-p35 rabbit polyclonal (C-19).

Techniques: Fluorescence, Transfection, Microscopy

Journal: Molecular Vision

Article Title: Cyclin-dependent kinase 5 promotes the stability of corneal epithelial cell junctions

doi:

Figure Lengend Snippet: Rac activity is required for cell–cell adhesions in the human corneal limbal epithelial cells. The ShCDK5 human corneal limbal epithelial (HCLE) and olomoucine-treated cultures reduced Rac activity ( A ) leading to instability of the cell–cell adhesion junctions, while the HCLE cells have significantly higher Rac activity ( B ) Rho kinase activity was tested in the in ShCDK5 HCLE cells. C : Representative experiment showing a twofold increase in the Rho activity in confluent cultures of ShCDK5 and in olomoucine-treated cells when compared to the control Lamin A/C and HCLE. Significant (n=3; p≤0.05) increase in the Rho activity in the absence of Cdk5 with olomoucine ( D ) and decrease in Rac activity in the ShCdk5 and olomoucine, suggesting destabilization of cell–cell adhesions in the olomoucine and ShCDK5 cells, is represented in the graph ( B , D ). A cytoskeletal Rac modulator, IQGAP1, coimmunes precipitates with E-cadherin ( E , F ) in the ShCDK5 suggesting the internalization and degradation of E-cadherin. Stable cell–cell adhesions in HCLE cells are marked by significant reduction in the interaction of IQGAP1 with E-cadherin. ( E , F ). Note: Rho or Rac activity was measured as individual values normalized with the tubulin/actin and ratio of active Rho or active Rac against total Rho or total Rac, respectively. IB=immunoblot; IP=immunoprecipitation.

Article Snippet: The following antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA): anti-Cdk5 mouse monoclonal (DC-17; sc-249), anti-Cdk5 rabbit polyclonal (C-8, sc-173), anti-pY15-Cdk5 (sc-12918), and anti-p35 rabbit polyclonal (C-19).

Techniques: Activity Assay, Stable Transfection, Western Blot, Immunoprecipitation

Journal: Molecular Vision

Article Title: Cyclin-dependent kinase 5 promotes the stability of corneal epithelial cell junctions

doi:

Figure Lengend Snippet: Rho activity mediates the downstream regulation of CDK5 in promoting cell–cell junction formation. Cell aggregates were measured with and without Ca 2+ and defined as particle size in μm 2 using NIH image J. In the presence of the C3 transferase, a Rho kinase inhibitor, the human corneal limbal epithelial (HCLE) cells form larger cell aggregates ( A ), and there is a reduction in the particle size in the presence of CDK5 and Rho kinase inhibitors suggesting the role of Cdk5 downstream signaling via the Rho kinase in the cell–cell junction formation ( C ). Olomoucine alone ( E ) reduces the formation of cell–cell aggregates than the control untreated ( G ) HCLE. Ratio of particle size=particle size (um) in the presence of calcium/ particle size (um) without calcium is represented in the graph ( I ). A – H are 20X images.

Article Snippet: The following antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA): anti-Cdk5 mouse monoclonal (DC-17; sc-249), anti-Cdk5 rabbit polyclonal (C-8, sc-173), anti-pY15-Cdk5 (sc-12918), and anti-p35 rabbit polyclonal (C-19).

Techniques: Activity Assay

Journal: Molecular immunology

Article Title: MiR-142-5p directly targets cyclin-dependent kinase 5-mediated upregulation of the inflammatory process in acquired middle ear cholesteatoma.

doi: 10.1016/j.molimm.2021.11.017

Figure Lengend Snippet: Fig. 2. The expression of miR-142-5p is negatively correlated with CDK5 expression. (A) The relative expression of miR-142-5p in AMEC epithelial tissues compared with that in control epithelial tissues was determined by qRT-PCR. (B) There was a significant negative correlation between miR-142-5p and CDK5. (C) Western blotting was used to determine the protein expression of CDK5 in LPS-induced HaCaT cells, with the expression of GAPDH used as a reference. (D) The relative protein level of CDK5 was assessed. (E, F)The relative expression of miR-142-5p and CDK5 was measured by qRT-PCR in HaCaT cells stimulated by LPS for different times. (G) Immunofluorescence analysis was performed in HaCaT cells with prolonged LPS stimulation (×400 magnification; CDK5: green; nucleus: blue; scale bar = 20 μm). Bar graphs show quantification of the results. Each value represents the mean ± SD of three independent experiments. Statistical significance was determined using the Student’s t-test. The groups of 6 h, 12 h, 24 h, 48 h and 72 h were compared to group 0 h. *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: A rabbit polyclonal anti-CDK5 antibody (Cat. No. sc-173, diluted 1:100 for IF) was obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA) and a mouse polyclonal anti-CDK5 antibody (Cat. No. BF0121, diluted 1:2,000 for Western blot analysis and diluted 1:100 for immunohistochemistry analysis) was purchased from Affinity Biosciences (Cincinnati, OH, USA).

Techniques: Expressing, Control, Quantitative RT-PCR, Western Blot, Immunofluorescence

Journal: Molecular immunology

Article Title: MiR-142-5p directly targets cyclin-dependent kinase 5-mediated upregulation of the inflammatory process in acquired middle ear cholesteatoma.

doi: 10.1016/j.molimm.2021.11.017

Figure Lengend Snippet: Fig. 3. The expression of inflammatory factors in AMEC and control epithelial tissuesispositively correlated with CDK5 in mRNA expression level. (A–E) QRT-PCR was used to identify the relative mRNA expression of inflammatory cytokines in AMEC and control epithelial tissues. (F–J) Positive correlations between CDK5 and inflammatory cytokines were identified. Bar graphs show quantification of the results. Each value represents the mean ± SD of three independent ex periments. Statistical significance was determined using the Student’s t-test. The groups of 6 h, 12 h, 24 h, 48 h and72 h were compared to group 0 h. *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: A rabbit polyclonal anti-CDK5 antibody (Cat. No. sc-173, diluted 1:100 for IF) was obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA) and a mouse polyclonal anti-CDK5 antibody (Cat. No. BF0121, diluted 1:2,000 for Western blot analysis and diluted 1:100 for immunohistochemistry analysis) was purchased from Affinity Biosciences (Cincinnati, OH, USA).

Techniques: Expressing, Control, Quantitative RT-PCR

Journal: Molecular immunology

Article Title: MiR-142-5p directly targets cyclin-dependent kinase 5-mediated upregulation of the inflammatory process in acquired middle ear cholesteatoma.

doi: 10.1016/j.molimm.2021.11.017

Figure Lengend Snippet: Fig. 6. MiR-142-5pinhibitsCDK5 expression through targeting CDK5. (A) The protein expression of CDK5 was determined by western blotting in HaCaT cells after si-CDK5, miR-142- 5p mimic, miR-142-5p inhibitor transfection, while siRNA, mimic and inhibitor was trans fected as a negative control. (B) CDK5 protein expression level was normalized to GAPDH. (C, D) The relative expression levels of miR-142-5p and CDK5 mRNA were determined by qRT-PCR in HaCaT cells after transfection. Bar graphs show quantification of the results. Each value represents the mean ± SD of three independent experiments. Statistical significance was deter mined using the Student’s t-test. NS: no signif icance, *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: A rabbit polyclonal anti-CDK5 antibody (Cat. No. sc-173, diluted 1:100 for IF) was obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA) and a mouse polyclonal anti-CDK5 antibody (Cat. No. BF0121, diluted 1:2,000 for Western blot analysis and diluted 1:100 for immunohistochemistry analysis) was purchased from Affinity Biosciences (Cincinnati, OH, USA).

Techniques: Expressing, Western Blot, Transfection, Negative Control, Quantitative RT-PCR

Journal: Molecular immunology

Article Title: MiR-142-5p directly targets cyclin-dependent kinase 5-mediated upregulation of the inflammatory process in acquired middle ear cholesteatoma.

doi: 10.1016/j.molimm.2021.11.017

Figure Lengend Snippet: Fig. 5. MiR-142-5p exerts an inhibitory ef fect on CDK5by directly targeting its 3’-UTR region. (A)The map of wild type CDK5 vector plasmids. (B)The map of mutant type CDK5 vector plasmids. (C) The 3’-UTR region of CDK5 may be a binding site of miR142-5p, as pre dicted in TargetScan. (D) Assay of the relative activity of luciferase in HEK293T cells. Bar graphs show quantification of the results. Each value represents the mean ± SD of three inde pendent experiments. Statistical significance was determined using the Student’s t-test. *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: A rabbit polyclonal anti-CDK5 antibody (Cat. No. sc-173, diluted 1:100 for IF) was obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA) and a mouse polyclonal anti-CDK5 antibody (Cat. No. BF0121, diluted 1:2,000 for Western blot analysis and diluted 1:100 for immunohistochemistry analysis) was purchased from Affinity Biosciences (Cincinnati, OH, USA).

Techniques: Plasmid Preparation, Mutagenesis, Binding Assay, Activity Assay, Luciferase

Journal: Molecular immunology

Article Title: MiR-142-5p directly targets cyclin-dependent kinase 5-mediated upregulation of the inflammatory process in acquired middle ear cholesteatoma.

doi: 10.1016/j.molimm.2021.11.017

Figure Lengend Snippet: Fig. 8. MiR-142-5p targets the CDK5-mediated upregulation of the expression of inflammatory cytokines in mRNA level. (A-E) QRT-PCR was performed to confirm the relative expression of miR-142-5p and CDK5 mRNA in HaCaT cells after transfection, while siRNA and inhibitor were transfected as negative control. Bar graphs show quantification of the results. Each value represents the mean ± SD of three independent experiments. Statistical significance was determined using the Student’s t-test. *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: A rabbit polyclonal anti-CDK5 antibody (Cat. No. sc-173, diluted 1:100 for IF) was obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA) and a mouse polyclonal anti-CDK5 antibody (Cat. No. BF0121, diluted 1:2,000 for Western blot analysis and diluted 1:100 for immunohistochemistry analysis) was purchased from Affinity Biosciences (Cincinnati, OH, USA).

Techniques: Expressing, Quantitative RT-PCR, Transfection, Negative Control

Journal: Molecular immunology

Article Title: MiR-142-5p directly targets cyclin-dependent kinase 5-mediated upregulation of the inflammatory process in acquired middle ear cholesteatoma.

doi: 10.1016/j.molimm.2021.11.017

Figure Lengend Snippet: Fig. 7. MiR-142-5p targets the CDK5-mediated upregulation of the expression of inflammatory cytokines by Elisa test. (A-E) The expression of inflammatory cytokines was detected by ELISA test in HaCaT cells transfected with LPS, si-CDK5 or miR-142-5p inhibitor, while siRNA and inhibitor were transfected as negative control. Bar graphs show quantification of the results. Each value represents the mean ± SD of three independent experiments. Statistical significance was determined using the Student’s t-test. NS: no significance, *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: A rabbit polyclonal anti-CDK5 antibody (Cat. No. sc-173, diluted 1:100 for IF) was obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA) and a mouse polyclonal anti-CDK5 antibody (Cat. No. BF0121, diluted 1:2,000 for Western blot analysis and diluted 1:100 for immunohistochemistry analysis) was purchased from Affinity Biosciences (Cincinnati, OH, USA).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Transfection, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: Role of cyclin-dependent kinase 5 in early brain injury following experimental subarachnoid hemorrhage

doi: 10.3892/etm.2021.11070

Figure Lengend Snippet: Expression levels of Cdk5, Cdk5-pTyr15 and p25 over time following SAH. (A) Representative autoradiograms of the expression levels of Cdk5, Cdk5-pTyr15 and p25 in the temporal cortex following SAH. Quantitative analysis of the western blotting results; (B) Cdk5 protein levels significantly increased at 12 h and on day 1 after SAH; (C) Cdk5-pTyr15 protein levels increased at 6 and 12 h and on day 1 after SAH; and (D) p25 protein levels increased on days 1 and 3 after SAH. (E) Quantified ratio of Cdk5-pTyr15/Cdk5 expression. * P<0.05, ** P<0.01 compared with the sham group. Cdk5, cyclin-dependent kinase 5; Cdk5-pTyr15, Cdk5 phosphorylated at Tyr15; SAH, subarachnoid hemorrhage.

Article Snippet: In total, 20 µg proteins were then separated by 10% SDS-PAGE and transferred onto a polyvinylidene fluoride membrane, which was blocked for 30 min in 5% non-fat milk in 1X TBS-0.1% Tween at 37˚C and then incubated with primary antibodies at 37˚C for Cdk5 (1:1,000, cat. no. ab40773; Abcam), Cdk5-pTyr15 (1:1,000; cat. no. AP55874PU-N; OriGene Technologies, Inc.), p25 (1:1,000; cat. no. ab125653; Abcam) and β-actin (1:1,000; cat. no. ab8226; Abcam) overnight.

Techniques: Expressing, Western Blot

Journal: Experimental and Therapeutic Medicine

Article Title: Role of cyclin-dependent kinase 5 in early brain injury following experimental subarachnoid hemorrhage

doi: 10.3892/etm.2021.11070

Figure Lengend Snippet: Double-immunofluorescence staining of Cdk5 after SAH. Cdk5 (green), NeuN (red) and GFAP (red) in the sham and day 1 post-SAH groups; nuclei were counterstained with DAPI (blue). Overlapping images show that Cdk5 was expressed in neurons and astrocytes in both the sham and SAH groups. Cdk5 was mainly expressed in the neuronal cytoplasm in the sham group; however, Cdk5 translocated to the nucleus after SAH (white arrows). Moreover, enhanced Cdk5 immunoreactivity was detected in the astrocytes of the SAH group compared with those of the sham group after SAH (scale bars, 20 µm). Cdk5, cyclin-dependent kinase 5; NeuN, neuronal nuclei; GFAP, glial fibrillary acidic protein; SAH, subarachnoid hemorrhage.

Article Snippet: In total, 20 µg proteins were then separated by 10% SDS-PAGE and transferred onto a polyvinylidene fluoride membrane, which was blocked for 30 min in 5% non-fat milk in 1X TBS-0.1% Tween at 37˚C and then incubated with primary antibodies at 37˚C for Cdk5 (1:1,000, cat. no. ab40773; Abcam), Cdk5-pTyr15 (1:1,000; cat. no. AP55874PU-N; OriGene Technologies, Inc.), p25 (1:1,000; cat. no. ab125653; Abcam) and β-actin (1:1,000; cat. no. ab8226; Abcam) overnight.

Techniques: Double Immunofluorescence Staining

Journal: Experimental and Therapeutic Medicine

Article Title: Role of cyclin-dependent kinase 5 in early brain injury following experimental subarachnoid hemorrhage

doi: 10.3892/etm.2021.11070

Figure Lengend Snippet: Effects of roscovitine on Cdk5-pTyr15 expression. (A) Cdk5-pTyr15 expression, as demonstrated by western blotting. (B) Quantitative western blotting results for Cdk5-pTyr15/β-actin expression. (C) Quantitative western blotting results for Cdk5-pTyr15/Cdk5. Roscovitine (100 µg) treatment significantly inhibited SAH-induced Cdk5-pTyr15 upregulation. ** P<0.01 compared with the sham + vehicle group; ## P<0.01 compared with the SAH + vehicle group. Cdk5-pTyr15. SAH, subarachnoid hemorrhage; Cdk5-pTyr15, Cdk5 phosphorylated at Tyr15.

Article Snippet: In total, 20 µg proteins were then separated by 10% SDS-PAGE and transferred onto a polyvinylidene fluoride membrane, which was blocked for 30 min in 5% non-fat milk in 1X TBS-0.1% Tween at 37˚C and then incubated with primary antibodies at 37˚C for Cdk5 (1:1,000, cat. no. ab40773; Abcam), Cdk5-pTyr15 (1:1,000; cat. no. AP55874PU-N; OriGene Technologies, Inc.), p25 (1:1,000; cat. no. ab125653; Abcam) and β-actin (1:1,000; cat. no. ab8226; Abcam) overnight.

Techniques: Expressing, Western Blot