Journal: Oncology reports

Article Title: Intermittent low dose irradiation enhances the effectiveness of radio- and chemo-therapy for human colorectal adenocarcinoma cell line HT-29.

doi: 10.3892/or.2017.5679

Figure Lengend Snippet: Figure 5. Intermittent LDIR activates ERK and p38MAPK pathways in 5-FU chemotherapy. In order to inhibit the function of ERK and p38MAPK, 10 µM PD98059 and SB203580 were added to the cell culture media at 2 h prior to 5-FU chemotherapy. (A) Western blotting of the MEK/ERK pathway genes in HT-29 cells. β-actin was used as the internal control for blotting reaction. (B) Relative band density of western blotting. (C) Western blotting of the p38MAPK pathway genes in HT-29 cells. β-actin was used as the internal control for blotting reaction. (D) Relative band density of western blotting. (E) ERK pathway inhibitors could not reverse the intermittent LDIR induced cell proliferation inhibition. (F) p38MAPK pathway inhibitors neutralized the intermittent LDIR induced cell proliferation inhibition. L, LDIR; IL, intermittent LDIR; NS, not significant. **p<0.01 as compared with the single dose LDIR group.

Article Snippet: The blots were blocked with 5% non-fat milk in TBST at 37 ̊C for 1 h. Then, blots were probed with monoclonal antibodies against ATM (1:500, Abcam, Shanghai, China), p53 (1:1,500, DO7, Santa Cruz Biotechnology, Santa Cruz, CA, USA), p21 (1:500, C-19, Santa Cruz Biotechnology), cyclin A (1:500, Santa Cruz Biotechnology), CDK2 (1:500, Santa Cruz Biotechnology), caspase-3 (1:1,000, Cell Signaling Technology, Beijing, China), cleaved caspase-3 (1:1,000, Cell Signaling Technology), ERK/phos-ERK (1:1,000, Cell Signaling Technology), p38/phos-p38 (1:1,000, Cell Signaling Technology) and β-actin (1:3,000, Santa Cruz Biotechnology) at 4 ̊C overnight.

Techniques: Cell Culture, Western Blot, Control, Inhibition

Journal: Poultry Science

Article Title: PKA and PKC signaling pathways mediate vitamin D₃-regulated intestinal phosphorus absorption in broiler chickens

doi: 10.1016/j.psj.2026.106762

Figure Lengend Snippet: Effects of dietary vitamin D 3 supplementation on jejunal phosphorylation levels of PKA, PKC, PI3K, p38MAPK, and ERK in broiler chickens (19 d) (Experiment 1). The control and vitamin D 3 diets contained 0 and 1000 IU/kg vitamin D 3 , respectively. (a) p-PKA/t-PKA; (b) p-PKC/t-PKC; (c) p-PI3K/t-PI3K; (d) p-p38MAPK/t-p38MAPK; (e) p-ERK/t-ERK. Protein abundance values are means ± SD of four replicates per treatment (1 broiler per replicate) (n = 4). Values with different letters differ between treatments ( P < 0.05).

Article Snippet: PVDF membranes were incubated with primary antibodies against NaPi-IIb (1:1000, A9460; ABclonal, Wuhan, China), phosphorylated PKA (p-PKA; 1:1000, 5661S), phosphorylated PI3K (p-PI3K; 1:1000, 4228), phosphorylated ERK (p-ERK; 1:1000, 9101S), phosphorylated p38MAPK (p-p38MAPK; 1:1000, 9216S), total PKA (t-PKA; 1:1000, 5842S) (Cell Signaling Technology, Boston, USA), total PI3K (t-PI3K; 1:10000, 60225-1-Ig), total ERK (t-ERK; 1:8000, 11257-1-AP-50), total p38MAPK(t-p38MAPK; 1:1000, 14064-1-AP), total PKC (t-PKC; 1:1000, 21991-1-AP), phosphorylated PKC (p-PKC; 1:5000, 29123-1-AP), and GAPDH (1:10000, 60004-1-Ig) (Proteintech, Wuhan, China).

Techniques: Phospho-proteomics, Control, Quantitative Proteomics

Journal: Poultry Science

Article Title: PKA and PKC signaling pathways mediate vitamin D₃-regulated intestinal phosphorus absorption in broiler chickens

doi: 10.1016/j.psj.2026.106762

Figure Lengend Snippet: Effects of the PKA inhibitor H-89 on jejunal phosphorylation levels of PKC, PI3K, p38MAPK, and ERK in broiler chickens (10–15 d) (Experiment 2). (a) p-PKC/t-PKC; (b) p-PI3K/t-PI3K; (c) p-p38MAPK/t-p38MAPK; (d) p-ERK/t-ERK. Protein abundance values are means ± SD of four replicates per treatment (1 broiler per replicate) (n = 4). Values with different letters differ between treatments ( P < 0.05).

Article Snippet: PVDF membranes were incubated with primary antibodies against NaPi-IIb (1:1000, A9460; ABclonal, Wuhan, China), phosphorylated PKA (p-PKA; 1:1000, 5661S), phosphorylated PI3K (p-PI3K; 1:1000, 4228), phosphorylated ERK (p-ERK; 1:1000, 9101S), phosphorylated p38MAPK (p-p38MAPK; 1:1000, 9216S), total PKA (t-PKA; 1:1000, 5842S) (Cell Signaling Technology, Boston, USA), total PI3K (t-PI3K; 1:10000, 60225-1-Ig), total ERK (t-ERK; 1:8000, 11257-1-AP-50), total p38MAPK(t-p38MAPK; 1:1000, 14064-1-AP), total PKC (t-PKC; 1:1000, 21991-1-AP), phosphorylated PKC (p-PKC; 1:5000, 29123-1-AP), and GAPDH (1:10000, 60004-1-Ig) (Proteintech, Wuhan, China).

Techniques: Phospho-proteomics, Quantitative Proteomics

Journal: Poultry Science

Article Title: PKA and PKC signaling pathways mediate vitamin D₃-regulated intestinal phosphorus absorption in broiler chickens

doi: 10.1016/j.psj.2026.106762

Figure Lengend Snippet: Effects of the PKC inhibitor staurosporine on duodenal phosphorylation levels of PKA, PI3K, p38MAPK, and ERK in broiler chickens (13 d) (Experiment 3). (a) p-PKA/t-PKA; (b) p-PI3K/t-PI3K; (c) p-p38MAPK/t-p38MAPK; (d) p-ERK/t-ERK. Protein abundance values are means ± SD of four replicates per treatment (1 broiler per replicate) (n = 4). Values with different letters differ between treatments ( P < 0.05).

Article Snippet: PVDF membranes were incubated with primary antibodies against NaPi-IIb (1:1000, A9460; ABclonal, Wuhan, China), phosphorylated PKA (p-PKA; 1:1000, 5661S), phosphorylated PI3K (p-PI3K; 1:1000, 4228), phosphorylated ERK (p-ERK; 1:1000, 9101S), phosphorylated p38MAPK (p-p38MAPK; 1:1000, 9216S), total PKA (t-PKA; 1:1000, 5842S) (Cell Signaling Technology, Boston, USA), total PI3K (t-PI3K; 1:10000, 60225-1-Ig), total ERK (t-ERK; 1:8000, 11257-1-AP-50), total p38MAPK(t-p38MAPK; 1:1000, 14064-1-AP), total PKC (t-PKC; 1:1000, 21991-1-AP), phosphorylated PKC (p-PKC; 1:5000, 29123-1-AP), and GAPDH (1:10000, 60004-1-Ig) (Proteintech, Wuhan, China).

Techniques: Phospho-proteomics, Quantitative Proteomics

Journal: Journal of Biological Chemistry

Article Title: Differential Interactions between Transforming Growth Factor-β3/TβR1, TAB1, and CD2AP Disrupt Blood-Testis Barrier and Sertoli-Germ Cell Adhesion

doi: 10.1074/jbc.m601618200

Figure Lengend Snippet: FIGURE 5. Changes in different target proteins, including MAP kinases in testes during TGF-3-induced BTB and AJ disruption in the testis. A, immunoblot analysis to assess the changes of protein levels after TGF-3 treatment in the rat testis. These include the representative integral junction membrane proteins and their adaptors at the BTB and apical ES, the three MAP kinases, and Smad2/3 and their corresponding phosphorylated (activated) forms (panel a). Parallel immunoblot analysis of these proteins in the testes of adult rats received vehicle control (panel b). B, densitometric scanned data of immunoblots such as those shown in A. Each bar is the mean S.D. of three independents sets of samples from different experiments. C, intrinsic kinase activity of ERK (substrate, Elk-1) and p38 (substrate, ATF-2) (top panel), and these data were also summarized in the bottom panel (n 3). ns, not significantly different by ANOVA; *, p 0.05; **, p 0.01; h, hour; d, day; C, control; V, vehicle control; Elk-1, E26-like protein 1; ATF-2, activating transcription factor 2.

Article Snippet: Intrinsic ERK and p38 MAPK assays were performed using specific p-ERK and p38 MAPK assay kits (Cell Signaling) with the corresponding fusion proteins Elk-1 and ATF-2.

Techniques: Disruption, Western Blot, Membrane, Control, Activity Assay

Journal: Journal of Biological Chemistry

Article Title: Differential Interactions between Transforming Growth Factor-β3/TβR1, TAB1, and CD2AP Disrupt Blood-Testis Barrier and Sertoli-Germ Cell Adhesion

doi: 10.1074/jbc.m601618200

Figure Lengend Snippet: FIGURE 8. A schematic drawing that illustrates the association between CD2AP, TAB1, and TR1 (type 1 receptor for TGF-) can selectively disrupt BTB and Sertoli- germ cell adhesion or Sertoli-germ cell adhesion alone. This schematic drawing sum- marizes the results of the experiments reported herein. It illustrates that differential interactions of TAB1 and CD2AP with the TGF-3-TR1 complex in the seminiferous epithelium can activate downstream signal transducers p38 and/or ERK. For instance, the association of the TGF-3-TR1 complex with adaptors TAB1 and CD2AP activates both p38 and/or ERK, perturbing BTB and AJ. However, if this protein complex only associates with CD2AP but not TAB1, it activates only ERK, selectively perturbing AJ without compromising the BTB.

Article Snippet: Intrinsic ERK and p38 MAPK assays were performed using specific p-ERK and p38 MAPK assay kits (Cell Signaling) with the corresponding fusion proteins Elk-1 and ATF-2.

Techniques:

Journal: Scientific Reports

Article Title: Deubiquitinating enzyme USP42 promotes breast cancer progression by inhibiting JNK/p38-mediated apoptosis

doi: 10.1038/s41598-025-20573-x

Figure Lengend Snippet: USP42 downregulation increases apoptosis and regulates JNK and p38 phosphorylation in breast cancer cells. ( A , B ) Flow cytometric analysis of apoptotic cell numbers following USP42 downregulation in MCF-7 ( A ) and MDA-MB-231 ( B ) cells. ( C ) Western blot analysis of cleaved-caspase3, Bcl-2 and Bax protein levels in MCF-7 and MDA-MB-231 cells. β-actin served as an internal reference. ( D ) Western blotting analysis of p-p38, p38, JNK and p-JNK protein expression in MCF-7 and MDA-MB-231 cells. β-actin served as an internal reference.

Article Snippet: Specific information for the antibodies used in the experiment is as follows: USP42 (1:500, sc-390604, Santa Cruz Biotechnology, Inc., USA), JNK (9252, Cell Signaling Technology, 1:1000), p-JNK (9251, Cell Signaling Technology, 1:1000), p38 (9212, Cell Signaling Technology, 1:1000), p-p38 (9211, Cell Signaling Technology, 1:1000), β-actin (sc-47778, Santa Cruz Biotechnology, 1:1000).

Techniques: Phospho-proteomics, Western Blot, Expressing

Journal: Scientific Reports

Article Title: Deubiquitinating enzyme USP42 promotes breast cancer progression by inhibiting JNK/p38-mediated apoptosis

doi: 10.1038/s41598-025-20573-x

Figure Lengend Snippet: USP42 downregulation inhibits cell growth via the JNK/p38 signaling pathway. ( A - D ) Proliferation of MCF-7 and MDA-MB-231 cells transfected with shUSP42-2# or negative control, as assessed by CCK-8 assay ( A , C ) and clonogenic assay ( B , D ). The cells were treated with SP600125 (JNK inhibitor), SB203580 (p38 MAPK inhibitor), or DMSO (negative control). ( ns : no significance , *: p < 0.05, **: p < 0.01, ***: p < 0.001).

Article Snippet: Specific information for the antibodies used in the experiment is as follows: USP42 (1:500, sc-390604, Santa Cruz Biotechnology, Inc., USA), JNK (9252, Cell Signaling Technology, 1:1000), p-JNK (9251, Cell Signaling Technology, 1:1000), p38 (9212, Cell Signaling Technology, 1:1000), p-p38 (9211, Cell Signaling Technology, 1:1000), β-actin (sc-47778, Santa Cruz Biotechnology, 1:1000).

Techniques: Transfection, Negative Control, CCK-8 Assay, Clonogenic Assay

Journal: Scientific Reports

Article Title: Deubiquitinating enzyme USP42 promotes breast cancer progression by inhibiting JNK/p38-mediated apoptosis

doi: 10.1038/s41598-025-20573-x

Figure Lengend Snippet: USP42 downregulation increases breast cancer cells apoptosis via the JNK/p38 signaling pathway. ( A , D ) Flow cytometry analysis of apoptotic MCF-7 and MDA-MB-231 cells transfected with shUSP42-2# or the negative control, treated with SP600125 ( A ), SB203580 ( D ) or DMSO. ( B , C ) Western blot analysis of cleaved caspase 3, Bcl-2, Bax, p-p38 MAPK, p38 MAPK, JNK and p-JNK expression in MCF-7 and MDA-MB-231 cells transfected with shUSP42-2# or negative control, treated with SP600125 ( B ), SB203580 ( C ) or DMSO. β-actin served as an internal reference. (*: p < 0.05, **: p < 0.01, ***: p < 0.001).

Article Snippet: Specific information for the antibodies used in the experiment is as follows: USP42 (1:500, sc-390604, Santa Cruz Biotechnology, Inc., USA), JNK (9252, Cell Signaling Technology, 1:1000), p-JNK (9251, Cell Signaling Technology, 1:1000), p38 (9212, Cell Signaling Technology, 1:1000), p-p38 (9211, Cell Signaling Technology, 1:1000), β-actin (sc-47778, Santa Cruz Biotechnology, 1:1000).

Techniques: Flow Cytometry, Transfection, Negative Control, Western Blot, Expressing

Journal: Scientific Reports

Article Title: Deubiquitinating enzyme USP42 promotes breast cancer progression by inhibiting JNK/p38-mediated apoptosis

doi: 10.1038/s41598-025-20573-x

Figure Lengend Snippet: USP42 promotes breast cancer tumor growth in vivo . ( A ) Schematic representation of in vivo experimental design. ( B ) Subcutaneous administration of MDA-MB-231 cells stably transfected with shUSP42-2# or the control vector into nude mice ( n = 4 per group). ( C ) Representative images of xenograft tumors in nude mice ( n = 4 per group). ( D ) Tumor weights in the shUSP42 and control vector groups ( n = 4 per group). ( E ) Western blot analysis of USP42, p-p38 MAPK, p38 MAPK, JNK and p-JNK protein expression in tumor tissues of xenograft mouse model ( n = 4 per group). (Data are presented as mean ± SD. *** p < 0.001).

Article Snippet: Specific information for the antibodies used in the experiment is as follows: USP42 (1:500, sc-390604, Santa Cruz Biotechnology, Inc., USA), JNK (9252, Cell Signaling Technology, 1:1000), p-JNK (9251, Cell Signaling Technology, 1:1000), p38 (9212, Cell Signaling Technology, 1:1000), p-p38 (9211, Cell Signaling Technology, 1:1000), β-actin (sc-47778, Santa Cruz Biotechnology, 1:1000).

Techniques: In Vivo, Stable Transfection, Transfection, Control, Plasmid Preparation, Western Blot, Expressing

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Protein kinase D interaction with TLR5 is required for inflammatory signaling in response to bacterial flagellin.

doi: 10.4049/jimmunol.178.9.5735

Figure Lengend Snippet: FIGURE 6. Effects of PKC/PKD inhibitors on flagellin stimulated NF-B translocation and p38 acti- vation in Caco cells. Cells were pretreated with DMSO, 3 M Go¨6976, or 12 M Go¨6983 for 1 h before addi- tion of 1 g/ml FliC for 30 min. A, EMSA to measure NF-B nuclear migration in Caco-2 cells. Nuclear extracts were purified, bound to radiolabeled NF-B-binding oli- gonucleotide, and separated by PAGE. Densitometric measurement on autoradiographs of NF-B-shifted bands is from three separate experiments (top) shown as the percentage (SEM). Bands corresponding to the NF- B-bound probe are shown from one representative ex- periment (bottom). B, Western blot analysis of p38 MAPK activation after FliC or IL-1 stimulation (top) is shown. After stimulation with either 0.5 g/ml FliC or 10 ng/ml IL-1 for the indicated times, cells were lysed. Equal amounts of lysate proteins were separated by SDS-PAGE, blotted, and then probed sequentially with anti-phospho-p38 MAPK (T180/Y182) and with total p38. Densitometric analysis of signals from 0 and 30 min after stimulation is shown for three pooled ex- periments (bottom). The relative intensity corresponds to the ratio of signal to maximal signal in DMSO for each experiment after correcting for differences in total p38. , p 0.01 by Tukey HSD.

Article Snippet: The following Abs were used: anti-PK (Serotec), anti-PKD1, antiphospho-PKD1 (S916), anti-phospho-p38 MAPK (T180/Y182), antitotal p38 (Cell Signaling Technology), and anti-GAPDH (Fitzgerald Industries International).

Techniques: Translocation Assay, Migration, Binding Assay, Western Blot, Activation Assay, SDS Page

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Protein kinase D interaction with TLR5 is required for inflammatory signaling in response to bacterial flagellin.

doi: 10.4049/jimmunol.178.9.5735

Figure Lengend Snippet: FIGURE 7. Effects of pharmacological and shRNA-mediated PKD inhibition on IL-8 release from flagellin-stimulated HEK 293T cells. A, Pharma- cological PKD inhibition of secreted IL-8. TLR5-transfected cells were pretreated with DMSO, 3 M Go¨6976, or 12 M Go¨6983 for 1 h and stimulated with 1 g/ml FliC or 200 nM PDB for 6 h, and supernatants were assayed for IL-8 content by ELISA (n 4). Results are expressed as a ratio of transfected to DMSO-treated, FliC-stimulated controls in each experiment. Data shown are mean SD. B, Representative Western blots showing the effect of shRNA on native PKD1 expression in HEK 293T cells. Following transfection as shown, cell lysates were separated by SDS-PAGE, blotted, and probed with anti-PKD1 and either anti-GAPDH (bottom right) or anti-total p38 (bottom left) as a loading control. C, Effect of siRNA-mediated, endogenous PKD1 knockdown on IL-8 release after FliC stimulation. Cells were transfected with TLR5 and empty pSuper, pSuper expressing one of two PKD shRNA sequences (pSPKD1-1 or pSPKD1-2), or the former sequence scrambled (pSscr1-1). Transfected cells were stimulated for 6 h with 0.5 g/ml FliC or 10 ng/ml IL-1, and IL-8 release was determined by ELISA (n 4 for IL-1; n 10 for FliC). Results were standardized to IL-8 release with empty pSuper in each experiment. See text for statistics. D, shRNA-mediated PKD1 knockdown specifically reduces FliC-induced phosphorylation of p38 MAPK. Cells were stimulated with either 0.5 g/ml FliC or 10 ng/ml IL-1 for 0, 5, or 20 min, and equal amounts of lysate proteins were separated by SDS-PAGE. The gels were blotted and then probed with anti-phospho-p38 MAPK (T180/Y182), total p38, anti-PKD, and finally anti-V5 to detect TLR5. Blots are representative of two such experiments.

Article Snippet: The following Abs were used: anti-PK (Serotec), anti-PKD1, antiphospho-PKD1 (S916), anti-phospho-p38 MAPK (T180/Y182), antitotal p38 (Cell Signaling Technology), and anti-GAPDH (Fitzgerald Industries International).

Techniques: shRNA, Inhibition, Transfection, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, SDS Page, Control, Knockdown, Sequencing, Phospho-proteomics