Journal: Bioactive Materials

Article Title: Mechanically sensitized hydrogel microspheres trigger membrane receptor switch for cartilage repair

doi: 10.1016/j.bioactmat.2026.03.017

Figure Lengend Snippet: Mechanism of integrin receptor activation induced by membrane receptor switch. A) Schematic representation of Calpain protein activity verified using fluorophores. B) The effectiveness of the fluorescence method was verified based on different groups. C) Calpain protein activity in MSCs subjected to different treatments. D) The expression of CDK5 and p-Talin head detected by Western blot analysis. E) Quantitative analysis of the Western blot results for the CDK5 and p-Talin head protein. F) The expression of p-Talin head and p-Talin detected by Western blot analysis. G) Quantitative analysis of the Western blot results for the p-Talin head protein. H) Quantitative analysis of the Western blot results for the p-Talin protein. I) Schematic diagram of Talin head and integrin β interaction verified by SPR technique. J) The association-dissociation curves of Talin head and integrin β1. K) Integrin β1 activation was verified by flow cytometry. L) Quantitative analysis of fluorescence intensity by flow cytometry. M) The expression of FAK and p-FAK detected by Western blot analysis. N) The quantification of FAK phosphorylation levels. O) Mechanism of integrin receptor activation induced by membrane receptor switch. (∗ symbol represents comparison with HAMA group, # symbol represents comparison with OBNC group). (ns: non-significant, ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ## P < 0.01, ### P < 0.001).

Article Snippet: TRPC1 inhibitor (0.3 nM, Pico145, CAS No. 1628287-16-0), TRPM7 inhibitor (1.0 μM, VPC4, CAS No. 945604-76-2), TRPV2 inhibitor (5.0 μM, compound IV2-1, CAS No. 2242724-49-6), TRPM4 inhibitor (1.5 μM, CBA, CAS No. 351424-20-9), PIEZO1 inhibitor (2.5 μM, GsMTx4, CAS No. 1209500-46-8), integrin αvβ5 inhibitor (8.0 nM, Compound 12, CAS No.: 2615912-33-7), integrin αvβ1 inhibitor (0.3 nM, Compound C8, CAS No. 1689540-62-2), integrin α5β1 inhibitor (10 μM, ATN-161, 904763-27-5), and CDK5 inhibitor (5 nM, CDK5-IN-1, 2,639,540-19-3) were purchased from MCE Biotechnology Co., LTD. After the MSCs were treated, the cRGD solution was added at a concentration of 1:200 and incubated in the dark for 15 min, and the results were observed by fluorescence microscopy.

Techniques: Activation Assay, Membrane, Activity Assay, Fluorescence, Expressing, Western Blot, Flow Cytometry, Phospho-proteomics, Comparison

Journal: Biomolecules

Article Title: Targeting a Tau Kinase Cdk5, Cyclin-Dependent Kinase: A Blood-Based Diagnostic Marker and Therapeutic Earmark for Alzheimer’s Disease

doi: 10.3390/biom15101365

Figure Lengend Snippet: Box plot of serum level of ( A ) Cdk5 and ( B ) Mcl1 in AD, MCI and GC. mRNA expression levels of ( C ) Cdk5 and ( D ) Mcl1 in blood samples of AD, MCI and GC subjects using RT-PCR. Cyclin-Dependent Kinase 5 (Cdk5), Myeloid Cell Leukemia 1 (Mcl1), Alzheimer’s Disease (AD), Mild Cognitive Impairment (MCI) and Geriatric Control (GC). Experiments were performed in triplicate, and data is presented as mean ± SD (serum level) and mean ± SEM (mRNA level) [ n = 61 (AD), 55 (MCI) and 57 (GC)]. One-way ANOVA followed by post hoc comparison using the Bonferroni test was used to compare between groups. Significantly different at; *** p < 0.0001.

Article Snippet: The following primary antibodies, mouse IgG against human Cdk5 (Santa Cruz Biotechnology Inc., Dallas, TX, USA, cat. no. Sc6247) and rabbit IgG against human Mcl1 (Cell Signaling Technologies, cat. no. CST-45725), were immobilized on the different flow cells of a CM5 sensor chip employing an amine coupling kit, following the manufacturer’s protocol (GE Healthcare, Uppsala, Sweden).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Control, Comparison

Journal: Biomolecules

Article Title: Targeting a Tau Kinase Cdk5, Cyclin-Dependent Kinase: A Blood-Based Diagnostic Marker and Therapeutic Earmark for Alzheimer’s Disease

doi: 10.3390/biom15101365

Figure Lengend Snippet: Receiver-operating characteristic (ROC) curve of Cdk5: ( A ) AD vs. GC, ( B ) AD vs. MCI, ( C ) MCI vs. GC, and Mcl1 ( D ) AD vs. GC, ( E ) AD vs. MCI, ( F ) MCI vs. GC, determining area under curve (AUC), cutoff values, sensitivity and specificity. Alzheimer’s Disease (AD = 61), Mild Cognitive Impairment (MCI = 55), Geriatric Control (GC = 57), Cyclin-Dependent Kinase 5 (Cdk5), and Myeloid Cell Leukemia 1 (Mcl1).

Article Snippet: The following primary antibodies, mouse IgG against human Cdk5 (Santa Cruz Biotechnology Inc., Dallas, TX, USA, cat. no. Sc6247) and rabbit IgG against human Mcl1 (Cell Signaling Technologies, cat. no. CST-45725), were immobilized on the different flow cells of a CM5 sensor chip employing an amine coupling kit, following the manufacturer’s protocol (GE Healthcare, Uppsala, Sweden).

Techniques: Control

Journal: Biomolecules

Article Title: Targeting a Tau Kinase Cdk5, Cyclin-Dependent Kinase: A Blood-Based Diagnostic Marker and Therapeutic Earmark for Alzheimer’s Disease

doi: 10.3390/biom15101365

Figure Lengend Snippet: Scatter diagram showing correlation between serum protein concentration (ng/μL) and clinical scores: ( A ) Cdk5 vs. HMSE score, ( B ) Cdk5 vs. ACE III score, ( C ) Mcl1 vs. HMSE score, ( D ) Mcl1 vs. ACE III score by Pearson’s correlation analysis. Cyclin-Dependent Kinase 5 (Cdk5), Myeloid Cell Leukemia 1 (Mcl1), Hindi Mini-Mental State Examination (HMSE) and Addenbrooke’s Cognitive Examination (ACE III).

Article Snippet: The following primary antibodies, mouse IgG against human Cdk5 (Santa Cruz Biotechnology Inc., Dallas, TX, USA, cat. no. Sc6247) and rabbit IgG against human Mcl1 (Cell Signaling Technologies, cat. no. CST-45725), were immobilized on the different flow cells of a CM5 sensor chip employing an amine coupling kit, following the manufacturer’s protocol (GE Healthcare, Uppsala, Sweden).

Techniques: Protein Concentration

Journal: Biomolecules

Article Title: Targeting a Tau Kinase Cdk5, Cyclin-Dependent Kinase: A Blood-Based Diagnostic Marker and Therapeutic Earmark for Alzheimer’s Disease

doi: 10.3390/biom15101365

Figure Lengend Snippet: Ligand interaction diagram of roscovitine and YCWS showing hydrogen bonding (magenta line) with Cdk5 (PDB: IUNL) protein. ( A ) Roscovitine forms two hydrogen bonds with Cys83 and one with Asp86. ( B ) YCWS forms two hydrogen bonds with Asn144 and three each with Asp86 and Cys83. All representations are predicted by Glide SP docking in the Schrödinger suite.

Article Snippet: The following primary antibodies, mouse IgG against human Cdk5 (Santa Cruz Biotechnology Inc., Dallas, TX, USA, cat. no. Sc6247) and rabbit IgG against human Mcl1 (Cell Signaling Technologies, cat. no. CST-45725), were immobilized on the different flow cells of a CM5 sensor chip employing an amine coupling kit, following the manufacturer’s protocol (GE Healthcare, Uppsala, Sweden).

Techniques:

Journal: Biomolecules

Article Title: Targeting a Tau Kinase Cdk5, Cyclin-Dependent Kinase: A Blood-Based Diagnostic Marker and Therapeutic Earmark for Alzheimer’s Disease

doi: 10.3390/biom15101365

Figure Lengend Snippet: Binding mode of YCWS and roscovitine with CDK5 (PDB: IUNL). ( A ) Cdk5 is represented in cyan color. ( B , C ) The ATP-binding pocket between the N and C terminal lobes of the kinase is occupied by YCWS (green) in a similar way as it is occupied by roscovitine (magenta), as observed by the superimposition of roscovitine and peptide YCWS. The main structural elements of the kinase have been labeled. The figures were prepared using PYMOL software 3.1. ( D ) Sensorgram showing binding of different concentrations (0.4 µM, 0.6 µM and 1 µM) of YCWS on Ni–NTA sensor chip immobilized with Cdk5 protein by SPR analysis.

Article Snippet: The following primary antibodies, mouse IgG against human Cdk5 (Santa Cruz Biotechnology Inc., Dallas, TX, USA, cat. no. Sc6247) and rabbit IgG against human Mcl1 (Cell Signaling Technologies, cat. no. CST-45725), were immobilized on the different flow cells of a CM5 sensor chip employing an amine coupling kit, following the manufacturer’s protocol (GE Healthcare, Uppsala, Sweden).

Techniques: Binding Assay, Labeling, Software

Journal: Biomolecules

Article Title: Targeting a Tau Kinase Cdk5, Cyclin-Dependent Kinase: A Blood-Based Diagnostic Marker and Therapeutic Earmark for Alzheimer’s Disease

doi: 10.3390/biom15101365

Figure Lengend Snippet: Effect of peptide YCWS on the expression level of ( A ) various proteins (Cdk5, Mcl1, Aβ-amyloid, Tau, and pTau) in different experimental groups of the Aβ-treated SH-SY5Y cell line treated with three different concentrations of peptide P1 (2 µM), P2 (5 µM) and P3 (10 µM) in culture medium for 72 h at 37 °C. ( B ) Cdk5 mRNA in all group’s was further validated by RT-PCR. Experiments were performed in triplicate and data is represented as mean ± SD (protein level) and mean ± SEM (mRNA level). The p -value is calculated between the Aβ-treated group and the Aβ + 10 µM peptide-treated group using the Mann–Whitney test. *** p -value ≤ 0.0001.

Article Snippet: The following primary antibodies, mouse IgG against human Cdk5 (Santa Cruz Biotechnology Inc., Dallas, TX, USA, cat. no. Sc6247) and rabbit IgG against human Mcl1 (Cell Signaling Technologies, cat. no. CST-45725), were immobilized on the different flow cells of a CM5 sensor chip employing an amine coupling kit, following the manufacturer’s protocol (GE Healthcare, Uppsala, Sweden).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, MANN-WHITNEY

Journal: Journal of Alzheimer's Disease Reports

Article Title: Progesterone induction of tau phosphorylation during the differentiation of human embryonic stem cells into neuroectodermal rosettes

doi: 10.1177/25424823251370643

Figure Lengend Snippet: Differentiation of hESC towards a neural phenotype promotes tau expression and phosphorylation. Pluripotent H9 hESC were cultured and differentiated into neural lineages as described in the methods. Following collection of the lineages, these samples, together with M17 neuroblastoma cells, MEFs, and control and AD brain were quantitated for protein, and equal amounts of protein run on SDS-PAGE and the immunoblots probed using well characterized antibodies against (A) anti-tau-1 (clone PC1C6), (B) Oct3/4, nestin, P-tau (AT8), Cdk5, and (C) PHF-1. Molecular weight markers are shown on the left-hand side.

Article Snippet: Polyclonal antibodies were used to detect human Cdk5 (Stressgen Bioreagent Corp., BC, Canada) and GSK-3β (Biolegend, CA, USA).

Techniques: Expressing, Phospho-proteomics, Cell Culture, Control, SDS Page, Western Blot, Molecular Weight

Journal: International Journal of Molecular Sciences

Article Title: Roscovitine, a CDK Inhibitor, Reduced Neuronal Toxicity of mHTT by Targeting HTT Phosphorylation at S1181 and S1201 In Vitro

doi: 10.3390/ijms252212315

Figure Lengend Snippet: Kinase assays identified upstream kinases modifying the serine of HTT peptides in vitro. In vitro kinase assay was used to screen the kinase targeting HTT peptide bearing serine 1181, serine 1201, and serine 2653 of HTT. ( A ) Kinases targeting the serine 1181 of the HTT peptide. ( B ) Kinases targeting the serine 1201 of the HTT peptide. ( C ) Kinases targeting the serine 2653 of the HTT peptide. CDK1 is highlighted by blue rectangles and CDK5/p25 by red rectangles. RBC, red blood cell substrate. Dotted lines indicate the 5% cut-off of enzyme activity used for the kinase screening.

Article Snippet: Human CDK5 expressing plasmid was purchased from Addgene (Watertown, MA, USA; Addgene plasmid # 1346; a gift from Li-Huei Tsai).

Techniques: In Vitro, Kinase Assay, Activity Assay

Journal: International Journal of Molecular Sciences

Article Title: Roscovitine, a CDK Inhibitor, Reduced Neuronal Toxicity of mHTT by Targeting HTT Phosphorylation at S1181 and S1201 In Vitro

doi: 10.3390/ijms252212315

Figure Lengend Snippet: HTT peptides as potential as targets for CDKs. ( A ) The amino acid sequences surrounding serine of 1181, 1201, and 2653 of HTT from different species. S/T-P-x-K/R is a consensus site for both CDK1 and CDK5, which means that the amino acid of S or T followed by P-x-K/R could be phosphorylated by CDK1 and CDK5. (S = serine; T = threonine; P = proline; K = lysine; R = arginine; x is any amino acid). It is well conserved in vertebrates for S1181 and S1201 of the HTT sequence, but not S2653. ( B ) CDKs target serine 1181 of the HTT peptide. ( C ) CDKs target serine 1201 of HTT peptide. CDK1 is highlighted by blue rectangles and CDK5/p25 by red rectangles. RBC, red blood cell substrate. Dotted lines indicate the 1% cut-off of enzyme activity used for the kinase screening.

Article Snippet: Human CDK5 expressing plasmid was purchased from Addgene (Watertown, MA, USA; Addgene plasmid # 1346; a gift from Li-Huei Tsai).

Techniques: Sequencing, Activity Assay

Journal: International Journal of Molecular Sciences

Article Title: Roscovitine, a CDK Inhibitor, Reduced Neuronal Toxicity of mHTT by Targeting HTT Phosphorylation at S1181 and S1201 In Vitro

doi: 10.3390/ijms252212315

Figure Lengend Snippet: The effect of knocking down CDK5 on the phosphorylation of S1181 and S1201 of mHTT. HEK293 cells were co-transfected with a plasmid expressing full-length mutant huntingtin with 82Q (FL-82Q) and siRNA targeting human CDK5 for 24 h. The expression level of CDK5, phosphorylated S1181-HTT (p-S1181), and phosphorylated S1201-HTT (p-S1201) were detected by western blot. ( A ) The representative western blot for CDK5. ( B ) The quantification of CDK5 expression. ( C ) The representative western blot for p-S1181-HTT. ( D ) The quantification of p-S1181-HTT expression. ( E ) The representative western blot for p-S1201-HTT. ( F ) The quantification of p-S1201-HTT expression. The experiment was repeated by at least two different analysts. Each experiment had n = 3. One representative experiment is presented. 2166 is the anti-HTT antibody (MAB2166, Millipore). RNAi = pooled siRNA targeting human CDK5. A two-tailed Student t -test was used. * p < 0.05; ** p < 0.01.

Article Snippet: Human CDK5 expressing plasmid was purchased from Addgene (Watertown, MA, USA; Addgene plasmid # 1346; a gift from Li-Huei Tsai).

Techniques: Phospho-proteomics, Transfection, Plasmid Preparation, Expressing, Mutagenesis, Western Blot, Two Tailed Test

Journal: International Journal of Molecular Sciences

Article Title: Roscovitine, a CDK Inhibitor, Reduced Neuronal Toxicity of mHTT by Targeting HTT Phosphorylation at S1181 and S1201 In Vitro

doi: 10.3390/ijms252212315

Figure Lengend Snippet: The effect of CDK5 overexpression on S1181 of mHTT. HEK293 cells were co-transfected with HTT plasmid FL-82Q and a plasmid expressing human CDK5 for 24 h. The expression levels of CDK5, p-S1181, and p-S1201 were detected by western blot. ( A ) The representative western blot for CDK5, p-S1181, 2166, and MW1. ( B ) The quantification of CDK5 expression. ( C , D ) The quantification of p-S1181 expression. ( E ) The quantification of total huntingtin (MAB2166) expression. ( F ) The quantification of mutant huntingtin (MW1) expression. The experiment was repeated by at least two different analysts. Each experiment had n = 3. One representative experiment is presented. 2166 is the anti-HTT antibody (MAB2166). MW1 is the anti-HTT antibody that binds to mutant huntingtin (clone MW1). RNAi = siRNA targeting human CDK5. A two-tailed Student t -test was used. ** p < 0.01; *** p < 0.001.

Article Snippet: Human CDK5 expressing plasmid was purchased from Addgene (Watertown, MA, USA; Addgene plasmid # 1346; a gift from Li-Huei Tsai).

Techniques: Over Expression, Transfection, Plasmid Preparation, Expressing, Western Blot, Mutagenesis, Two Tailed Test

Journal: International Journal of Molecular Sciences

Article Title: Roscovitine, a CDK Inhibitor, Reduced Neuronal Toxicity of mHTT by Targeting HTT Phosphorylation at S1181 and S1201 In Vitro

doi: 10.3390/ijms252212315

Figure Lengend Snippet: CDK5 level altered cell toxicity induced by mHTT. The effect of CDK5 overexpression or knocking down was evaluated in mouse striatal cells with different poly Q (SThdh Q7/Q7 or SThdh Q111/Q111 ). Cells were transfected with either a plasmid expressing human CDK5 or pooled siRNA targeting mouse CDK5 for 24 h. Cell death was measured with CytoTox-Glo cytotoxicity assay kit (Promega). The experiment was repeated by at least two different analysts. Each experiment had n = 3. One representative experiment is presented. Two-way ANOVA with Tukey’s multiple comparation was used for analysis. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: Human CDK5 expressing plasmid was purchased from Addgene (Watertown, MA, USA; Addgene plasmid # 1346; a gift from Li-Huei Tsai).

Techniques: Over Expression, Transfection, Plasmid Preparation, Expressing, Cytotoxicity Assay

Journal: International Journal of Molecular Sciences

Article Title: Roscovitine, a CDK Inhibitor, Reduced Neuronal Toxicity of mHTT by Targeting HTT Phosphorylation at S1181 and S1201 In Vitro

doi: 10.3390/ijms252212315

Figure Lengend Snippet: Roscovitine reduced the phosphorylation of S1181 and S1201 of mHTT. Roscovitine reduced p-S1181-HTT and p-S1201-HTT in vitro by inhibiting CDK5 but not CDK1. ( A ) The representative western blot for p-S1181 in HEK293 cells transfected with a plasmid expressing FL-82Q and treated with or without roscovitine for 48 h. ( B ) The representative western blot for p-S1201 in HEK293 cells transfected with a plasmid expressing FL-82Q and treated with or without roscovitine for 48 h. ( C ) The representative western blot for CDK5 in HEK293 cells transfected with a plasmid expressing FL-82Q and treated with or without roscovitine for 48 h. ( D ) The representative western blot for CDK1 in HEK293 cells transfected with a plasmid expressing FL-82Q and treated with or without roscovitine for 48 h. ( E ) Quantification of p-S1181 in A. ( F ) Quantification of p-S1201 in B. ( G ) Quantification of CDK5 in C. ( H ) Quantification of CDK1 in D. The experiment was repeated by at least two different analysts. Each experiment had n = 3. One representative experiment is presented. Ros~ = Roscovitine. A two-tailed Student t -test was used. * p < 0.05; ** p < 0.01.

Article Snippet: Human CDK5 expressing plasmid was purchased from Addgene (Watertown, MA, USA; Addgene plasmid # 1346; a gift from Li-Huei Tsai).

Techniques: Phospho-proteomics, In Vitro, Western Blot, Transfection, Plasmid Preparation, Expressing, Two Tailed Test

Journal: International Journal of Molecular Sciences

Article Title: Roscovitine, a CDK Inhibitor, Reduced Neuronal Toxicity of mHTT by Targeting HTT Phosphorylation at S1181 and S1201 In Vitro

doi: 10.3390/ijms252212315

Figure Lengend Snippet: Roscovitine penetrated the mouse brain and inhibited CDK5 activity in the zQ175 HD mouse brain. Variant doses of roscovitine were injected into 4-month-old mice through intraperitoneal injection (IP). Plasma and brain samples were collected at different time points after injection. The concentration of roscovitine was measured by LCMS/MS. ( A ) Brain concentration of roscovitine at different time points after acute IP injection with variant doses. Each group has 4–5 mice. ( B ) The CDK5 activity in the brain of mice injected with roscovitine for three weeks. Two doses of roscovitine were injected into 4-month-old zQ175HD mice by IP daily for three weeks. Twenty-four hours after the last injection, the mouse brain samples were collected, and the CDK5 activity was measured using a commercial kit from Promega. Animal number is indicated in the bar graph for each group. R-25 = 25 mg/kg of roscovitine. R-50 = 50 mg/kg of roscovitine. One-way ANOVA with Fish’s LSD was used for analysis. * p < 0.05; ** p < 0.01.

Article Snippet: Human CDK5 expressing plasmid was purchased from Addgene (Watertown, MA, USA; Addgene plasmid # 1346; a gift from Li-Huei Tsai).

Techniques: Activity Assay, Variant Assay, Injection, Clinical Proteomics, Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: Roscovitine, a CDK Inhibitor, Reduced Neuronal Toxicity of mHTT by Targeting HTT Phosphorylation at S1181 and S1201 In Vitro

doi: 10.3390/ijms252212315

Figure Lengend Snippet: Kinase assays identified upstream kinases modifying the serine of HTT peptides in vitro. In vitro kinase assay was used to screen the kinase targeting HTT peptide bearing serine 1181, serine 1201, and serine 2653 of HTT. ( A ) Kinases targeting the serine 1181 of the HTT peptide. ( B ) Kinases targeting the serine 1201 of the HTT peptide. ( C ) Kinases targeting the serine 2653 of the HTT peptide. CDK1 is highlighted by blue rectangles and CDK5/p25 by red rectangles. RBC, red blood cell substrate. Dotted lines indicate the 5% cut-off of enzyme activity used for the kinase screening.

Article Snippet: Pooled siRNA targeting human and mouse CDK5 and CDK1 was purchased from Santa Cruz Biotechnology (Santa Cruz Biotechnology, Inc., Dallas, TX, USA).

Techniques: In Vitro, Kinase Assay, Activity Assay

Journal: International Journal of Molecular Sciences

Article Title: Roscovitine, a CDK Inhibitor, Reduced Neuronal Toxicity of mHTT by Targeting HTT Phosphorylation at S1181 and S1201 In Vitro

doi: 10.3390/ijms252212315

Figure Lengend Snippet: HTT peptides as potential as targets for CDKs. ( A ) The amino acid sequences surrounding serine of 1181, 1201, and 2653 of HTT from different species. S/T-P-x-K/R is a consensus site for both CDK1 and CDK5, which means that the amino acid of S or T followed by P-x-K/R could be phosphorylated by CDK1 and CDK5. (S = serine; T = threonine; P = proline; K = lysine; R = arginine; x is any amino acid). It is well conserved in vertebrates for S1181 and S1201 of the HTT sequence, but not S2653. ( B ) CDKs target serine 1181 of the HTT peptide. ( C ) CDKs target serine 1201 of HTT peptide. CDK1 is highlighted by blue rectangles and CDK5/p25 by red rectangles. RBC, red blood cell substrate. Dotted lines indicate the 1% cut-off of enzyme activity used for the kinase screening.

Article Snippet: Pooled siRNA targeting human and mouse CDK5 and CDK1 was purchased from Santa Cruz Biotechnology (Santa Cruz Biotechnology, Inc., Dallas, TX, USA).

Techniques: Sequencing, Activity Assay

Journal: International Journal of Molecular Sciences

Article Title: Roscovitine, a CDK Inhibitor, Reduced Neuronal Toxicity of mHTT by Targeting HTT Phosphorylation at S1181 and S1201 In Vitro

doi: 10.3390/ijms252212315

Figure Lengend Snippet: Roscovitine reduced the phosphorylation of S1181 and S1201 of mHTT. Roscovitine reduced p-S1181-HTT and p-S1201-HTT in vitro by inhibiting CDK5 but not CDK1. ( A ) The representative western blot for p-S1181 in HEK293 cells transfected with a plasmid expressing FL-82Q and treated with or without roscovitine for 48 h. ( B ) The representative western blot for p-S1201 in HEK293 cells transfected with a plasmid expressing FL-82Q and treated with or without roscovitine for 48 h. ( C ) The representative western blot for CDK5 in HEK293 cells transfected with a plasmid expressing FL-82Q and treated with or without roscovitine for 48 h. ( D ) The representative western blot for CDK1 in HEK293 cells transfected with a plasmid expressing FL-82Q and treated with or without roscovitine for 48 h. ( E ) Quantification of p-S1181 in A. ( F ) Quantification of p-S1201 in B. ( G ) Quantification of CDK5 in C. ( H ) Quantification of CDK1 in D. The experiment was repeated by at least two different analysts. Each experiment had n = 3. One representative experiment is presented. Ros~ = Roscovitine. A two-tailed Student t -test was used. * p < 0.05; ** p < 0.01.

Article Snippet: Pooled siRNA targeting human and mouse CDK5 and CDK1 was purchased from Santa Cruz Biotechnology (Santa Cruz Biotechnology, Inc., Dallas, TX, USA).

Techniques: Phospho-proteomics, In Vitro, Western Blot, Transfection, Plasmid Preparation, Expressing, Two Tailed Test