Journal: Molecules

Article Title: The Preventive Effect of Specific Collagen Peptides against Dexamethasone-Induced Muscle Atrophy in Mice

doi: 10.3390/molecules28041950

Figure Lengend Snippet: Effect of CP on myokine alterations. ( A – D ) The mRNA expression of myostatin ( A ), TGF-β ( B ), BDNF ( C ), and FNDC5 ( D ) in the muscles was assessed using real-time PCR. ( E , F ) Total Smad2/3, phospho-Smad2, total-AKT, phospho-AKT, and GAPDH levels in the muscles were assessed by immunoblotting. Total proteins were stained by Ponceau S. ImageJ was used to measure and quantify the band intensity. Arrow in ( E ) indicates p-Smad 2, and this band was used to quantify the p-Smad2 level. The p-Smad2 and p-AKT levels were normalized by Ponceau S staining. ( G ) Total and phospho-mTOR levels were analyzed by ELISA. The p-mTOR levels were normalized against the total mTOR levels. Data are presented as the means ± SD of values obtained from seven biological replicates. # p < 0.05, ## p < 0.01, DEX vs. control. # p < 0.05, ## p < 0.01, DEX + CP (0.25 and 0.5) vs. control. * p < 0.05, ** p < 0.01, DEX + CP (0.25 and 0.5) vs. DEX; CP, collagen peptides; DEX, dexamethasone.

Article Snippet: To quantify the activity of mTOR and p-mTOR, Pathscan sandwich ELISA kits (mTOR: CST#7974, p-mTOR: CST#7976) were used.

Techniques: Expressing, Muscles, Real-time Polymerase Chain Reaction, Western Blot, Staining, Enzyme-linked Immunosorbent Assay, Control

Journal: Molecules

Article Title: The Preventive Effect of Specific Collagen Peptides against Dexamethasone-Induced Muscle Atrophy in Mice

doi: 10.3390/molecules28041950

Figure Lengend Snippet: Anti-inflammatory activity of CP in muscle tissue. ( A ) Evans blue staining of the calf muscle. The blue region in the representative images indicates damaged muscle fibers. The Evans blue-positive area was quantified and is presented as a relative percentage of the control below the representative photos. This experiment was performed with three mice per group. ( B , C ) The mRNA expression of TNF-α and IL-1β in the muscles was assessed using real-time PCR. Data are presented as the means ± SD of values obtained from seven biological replicates. ( D ) The IL-1β levels in serum were examined using ELISA. Data are presented as the means ± SD of values obtained from three biological replicates. # p < 0.05, ## p < 0.01, DEX vs. control. # p < 0.05, ## p < 0.01, DEX + CP (0.25 and 0.5) vs. control. * p < 0.05, ** p < 0.01, DEX + CP (0.25 and 0.5) vs. DEX; CP, collagen peptides; DEX, dexamethasone.

Article Snippet: To quantify the activity of mTOR and p-mTOR, Pathscan sandwich ELISA kits (mTOR: CST#7974, p-mTOR: CST#7976) were used.

Techniques: Activity Assay, Staining, Control, Expressing, Muscles, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Developmental cell

Article Title: Mammalian SEPT2 is required for scaffolding nonmuscle myosin II and its kinases.

doi: 10.1016/j.devcel.2007.09.001

Figure Lengend Snippet: Figure 4. Dissociation of SEPT2 from Myosin II Alters Myosin II Activity and Increases the Number of Binucleated Cells (A) Representative western blots of CHO-K1 whole-cell lysates prepared after incubating cells with PBS, the TAT-CCT3-pt, or TAT-CCT5 for 1.5 hr. Each lysate was probed with antibodies against the indicated proteins. (B) Quantification of the western blots showing relative levels of myosin II, MYPT1, pMYPT1, SEPT2, SEPT9, MLC, pMLC, and ppMLC. Each exper- iment was done a minimum of three times, normalized to PBS transduction. The error bars reflect the standard deviation. *p < 0.05. (C) Representative western blots of CHO-K1 whole-cell lysates prepared after transfecting with nonspecific shRNAi (shRNAi Scramble) or shRNAi silencing SEPT2. Each lysate was probed with antibodies against the indicated proteins. (D) A representative recording of cell division shown over time depicting cells that were transduced with the TAT-CCT5 or TAT-CCT3-pt. Recordings were started at the beginning of karyokinesis. The number on each panel indicates the time elapsed from the starting point, expressed in minutes.

Article Snippet: Collected samples were probed for SEPT9 (1:1000; Surka et al., 2002), SEPT2 (1:1000), myosin II (1:1000), MYPT1 or pMYPT1 (1:1000; Upstate Biotechnology), and pMLC or ppMLC (1:1000; Cell Signaling Technology).

Techniques: Activity Assay, Western Blot, Transduction, Standard Deviation

Journal: The AAPS journal

Article Title: Pharmacodynamics of Cyclin D1 Degradation in Ovarian Cancer Xenografts with Repeated Oral SHetA2 Dosing.

doi: 10.1208/s12248-023-00874-7

Figure Lengend Snippet: Fig. 2 A PBPK/PD model for SHetA2 in tumor-bearing mice. The PBPK model includes 14 perfusion-limited tissue compartments, elimination of SHetA2 from liver (CLliv), intes- tine (CLGI) and plasma (CLPL), and oral absorption kinetics (ka, ka1, FAGG). The PD model is based on the SHetA2 concentra- tion in the tumor and depicts the stimulation of cyclin D1 deg- radation through three transit compartments (TC1-3). Cyclin D1 synthesis and degradation are depicted by the zero-order constant kin and the first-order constant kout, respectively. The initial effect E induced by drug concentration (Cdrug) is described by the maximum effect (Smax), the drug concen- tration for 50% effect (SC50), and the slope factor (γ). The transit compartments account for events such as cyclin D1 phosphorylation, ubiquitination, and proteasomal degradation, with τ denoting the transit time among compartments

Article Snippet: PathScan® Total Cyclin D1 Sandwich ELISA Kit was purchased from Cell Signaling Technology (Boston, MA).

Techniques: Clinical Proteomics, Concentration Assay, Phospho-proteomics, Ubiquitin Proteomics

Journal: The AAPS journal

Article Title: Pharmacodynamics of Cyclin D1 Degradation in Ovarian Cancer Xenografts with Repeated Oral SHetA2 Dosing.

doi: 10.1208/s12248-023-00874-7

Figure Lengend Snippet: Fig. 5 Dose- and time-depend- ent effects of SHetA2 on cyclin D1 in vitro. Cyclin D1 protein levels were quantified in cell lysates using both ELISA (a, c, e, and f), which are presented as mean ± SD, and western blot- ting (b, d, and g). Two human ovarian cancer cell lines, A2780 and SKOV3, were treated with varying concentrations of SHetA2 (1, 2.5, 5, 7.5, 10, 15 μM) for 24 h (a and b). Time- dependent changes in cyclin D were monitored at different time points (0, 2, 4, 6, 8, 12, 24 h) after treating SKOV3 cells with 2.5 or 10 μM SHetA2 (c and d). The recovery of drug-induced cyclin D1 reduction upon drug removal was assessed at a dose of 10 μM SHetA2 (e, f, and g). After A2780 or SKOV3 cells were treated with control or SHetA2 for 24 h, the drug- containing culture medium was removed, cultures were washed, and the cells were incubated with fresh medium for an additional 24 h. The levels of cyclin D1 were monitored at 4, 8, and 24 h after drug removal. Representative western blots for mortalin and cyclin D1, along with GAPDH or cyclophilin G as the loading control (b, d, and g). Statistical analyses were performed in GraphPad Prism using one-way ANOVA tests

Article Snippet: PathScan® Total Cyclin D1 Sandwich ELISA Kit was purchased from Cell Signaling Technology (Boston, MA).

Techniques: In Vitro, Enzyme-linked Immunosorbent Assay, Western Blot, Control, Incubation

Journal: The AAPS journal

Article Title: Pharmacodynamics of Cyclin D1 Degradation in Ovarian Cancer Xenografts with Repeated Oral SHetA2 Dosing.

doi: 10.1208/s12248-023-00874-7

Figure Lengend Snippet: Fig. 6 The time profiles of cyclin D1 after treatment with SHetA2 in vitro and in vivo. a Changes in cyclin D1 protein levels over time in SKOV3 treated with different SHetA2 concentrations (1–15 μM) dur- ing a 24-h incubation followed by drug removal (the data are from Fig. 5). Different time points (0, 2, 4, 6, 8, 12, 24 h) were monitored for the 2.5 and 10 μM groups, and only 24-h time point was measured for the other doses. The recovery of drug-induced cyclin D1 reduc- tion upon drug removal was evaluated at 10 μM by washing the drug- containing culture medium and incubating with fresh medium for an additional 24 h. b The time profile of cyclin D1 levels in SKOV3 tumors in mice treated with a single (light blue circle) and 7-day (blue circles) oral dose of 60 mg/kg SHetA2 once daily. The sym- bols represent the observed cyclin D1 protein levels in cancer cells or tumors, quantified using the ELISA method, with data presented as mean ± SD (n = 3). Both in vitro and in vivo data were used to estimate the PD model parameters, and the lines represent the best- fit curves. c Sensitivity analysis for each parameter. Sensitivity coef- ficients are determined through the division of changes in the model’s output by variations in the parameters

Article Snippet: PathScan® Total Cyclin D1 Sandwich ELISA Kit was purchased from Cell Signaling Technology (Boston, MA).

Techniques: In Vitro, In Vivo, Incubation, Enzyme-linked Immunosorbent Assay

Journal: The AAPS journal

Article Title: Pharmacodynamics of Cyclin D1 Degradation in Ovarian Cancer Xenografts with Repeated Oral SHetA2 Dosing.

doi: 10.1208/s12248-023-00874-7

Figure Lengend Snippet: Fig. 7 The simulated time pro- files of SHetA2 concentration (a) and cyclin D1 (b) in tumors in mice treated with different dosage regimens of SHetA2

Article Snippet: PathScan® Total Cyclin D1 Sandwich ELISA Kit was purchased from Cell Signaling Technology (Boston, MA).

Techniques: Concentration Assay

Journal: Nanomedicine : nanotechnology, biology, and medicine

Article Title: Chemosensitization of tumors via simultaneous delivery of STAT3 inhibitor and doxorubicin through HPMA copolymer-based nanotherapeutics with pH-sensitive activation.

doi: 10.1016/j.nano.2023.102730

Figure Lengend Snippet: Fig. 6 S3i and its derivatives S3iD2 and S3iD3 inhibit STAT3 phosphorylation. CT26

Article Snippet: Determination of STAT3 and pSTAT3 levels in cell lysates (50 μg of protein) was carried out using a PathScan Total STAT3 Sandwich ELISA kit and PathScan Phospho-STAT3 (Y705) Sandwich ELISA kit (both Cell Signaling Technology, USA), respectively, according to the manufacturer’s instructions.

Techniques: Phospho-proteomics

Journal: Acta pharmaceutica (Zagreb, Croatia)

Article Title: Insights into the mechanism of antiproliferative effects of primaquine-cinnamic acid conjugates on MCF-7 cells.

doi: 10.2478/acph-2018-0021

Figure Lengend Snippet: Fig. 5. The effects of test compounds on the levels of apoptosis signaling proteins. Results represent mean values ± SEM. Significant difference vs. control, *p < 0.05.

Article Snippet: Apoptosis assay: Apoptosis protein levels Apoptosis proteins were measured using a PathScan Apoptosis Multi-Target Sandwich ELISA Kit (Cell Signaling Technology, USA) according to the manufacturer’s instructions.

Techniques: Control