Journal: Life Science Alliance

Article Title: DOCK1 insufficiency disrupts trophoblast function and pregnancy outcomes via DUSP4-ERK pathway

doi: 10.26508/lsa.202302247

Figure Lengend Snippet: (A) Volcano plot analysis showing the differentially expressed genes; genes with log 2 foldchange > 1 and P -value < 0.05 are indicated in red; genes with the log 2 foldchange < −1 and P -value < 0.05 are indicated in blue; genes with the |log 2 foldchange| < 1 or P -value > 0.05 are indicated in grey. (B) ERK protein phosphorylation levels in JAR cells with DOCK1 repression or overexpression. (C) ERK phosphorylation levels in HTR-8 cells treated with 10 ng/ml EGF for 0, 30, and 60 min. (D) Knockdown efficacy of DUSP4 siRNA and DOCK1 expression after DUSP4 knockdown detected in HTR-8 cells by Western blotting. (E) 293T cells were transfected with ectopic Flag-DOCK1, Myc-DUSP4, and HA-UB for 48 h, and then treated with 20 μM MG132 for 6 h. The cell lysates were immunoprecipitated with Myc antibody and ubiquitination assays showed the DUSP4 protein ubiquitination levels. (F) mRNA expression of DUSP4 after HUWE1 knockdown in HTR-8 cells. (G) Knockdown efficiency of HUWE1 siRNA and DUSP4 protein abundance after HUWE1 knockdown detected in HTR-8 cells by Western blotting.

Article Snippet: DUSP4 , IF(1:200) , Proteintech , #66349-1-Ig.

Techniques: Phospho-proteomics, Over Expression, Knockdown, Expressing, Western Blot, Transfection, Immunoprecipitation, Ubiquitin Proteomics, Quantitative Proteomics

Journal: Life Science Alliance

Article Title: DOCK1 insufficiency disrupts trophoblast function and pregnancy outcomes via DUSP4-ERK pathway

doi: 10.26508/lsa.202302247

Figure Lengend Snippet: (A) DUSP4 mRNA expression in DOCK1-knockout HTR-8 cells. (B) DUSP4 protein levels in DOCK1-depleted HTR-8 cells (left). The protein levels were quantified (right). (C) Wound healing ability after DUSP4 inhibition by siRNA in DOCK1-knockout HTR-8 cells (left). Rate of wound closure was quantified (right). Scale bar: 200 μm. (D) Migration and invasion ability after DUSP4 inhibition in DOCK1 knockout HTR-8 cells (top). The number of migrating and invading cells was quantified (bottom). Scale bar: 100 μm. (E) ERK phosphorylation and metastasis-related protein levels after DUSP4 siRNA treatment in DOCK1-knockout HTR-8 cells. In (A, C, D), each group, n = 3 biological replicates (* P < 0.05; ** P < 0.01; unpaired two-tailed t test). In (B), each group, n = 4 biological replicates (*** P < 0.001; unpaired two-tailed t test).

Article Snippet: DUSP4 , IF(1:200) , Proteintech , #66349-1-Ig.

Techniques: Expressing, Knock-Out, Inhibition, Migration, Phospho-proteomics, Two Tailed Test

Journal: Life Science Alliance

Article Title: DOCK1 insufficiency disrupts trophoblast function and pregnancy outcomes via DUSP4-ERK pathway

doi: 10.26508/lsa.202302247

Figure Lengend Snippet: (A) DUSP4 protein levels in DOCK1-overexpressing HTR-8 cells. (B) Interaction of DOCK1 and DUSP4 detected by CO-IP in HEK293T cells transfected with ectopic Flag-DOCK1 and Myc-DUSP4. (C) Interaction of DOCK1 and DUSP4 in the cytoplasm observed by immunofluorescence staining with anti-DOCK1 (red) and anti-DUSP4 (green) in HTR-8 cells. Scale bar: 25 μm. (D) DOCK1 knockout HTR-8 cells were treated with 50 μg/ml cycloheximide (CHX) for 0, 1.5, 3, and 6 h, and DOCK1 and DUSP4 proteins were assayed by Western blot. (E) HTR-8 cells overexpressing DOCK1 were treated with 50 μg/ml CHX for 0, 1.5, 3, and 6 h, and Western blot was used to detect DOCK1 and DUSP4 proteins. (F) HTR-8 cells were treated with 50 μg/ml CHX alone or 50 μg/ml CHX and 20 μM MG132 for 6 h. (G) HTR-8 cells were treated with 50 μg/ml CHX alone or 50 μg/ml CHX and 10 μM CQ or 50 μg/ml CHX plus 1 mM 3-MA for 6 h. (H) DOCK1-overexpression and control HTR-8 cells were treated with 20 μM MG132 or left untreated for 6 h.

Article Snippet: DUSP4 , IF(1:200) , Proteintech , #66349-1-Ig.

Techniques: Co-Immunoprecipitation Assay, Transfection, Immunofluorescence, Staining, Knock-Out, Western Blot, Over Expression, Control

Journal: Life Science Alliance

Article Title: DOCK1 insufficiency disrupts trophoblast function and pregnancy outcomes via DUSP4-ERK pathway

doi: 10.26508/lsa.202302247

Figure Lengend Snippet: (A) DOCK1-knockout HTR-8 cells were transfected with HA-UB for 48 h and treated with 20 μM MG132 for 6 h. The cell lysates were immunoprecipitated with the DUSP4 antibody, and the ubiquitination assay showed the ubiquitination levels of DUSP4 protein. (B) HUWE1 protein levels after DOCK1 knockout or overexpression in HTR-8 cells. (C) After DOCK1 overexpression followed by HUWE1 knockdown, HTR-8 cells were treated with 50 μg/ml cycloheximide for 0, 1.5, 3, and 6 h, and Western blot was performed to detect DOCK1 and DUSP4 protein levels. (D) 293T cells were transfected with ectopic Flag-DOCK1, Myc-DUSP4, and HA-UB for 24 h, and then transfected with HUWE1 siRNA for 48 h. Finally, 20 μM MG132 was added and the protein was extracted after 6 h of incubation. The cell lysates were immunoprecipitated with Myc antibody and the ubiquitination assay showed DUSP4 protein ubiquitination levels.

Article Snippet: DUSP4 , IF(1:200) , Proteintech , #66349-1-Ig.

Techniques: Knock-Out, Transfection, Immunoprecipitation, Ubiquitin Proteomics, Over Expression, Knockdown, Western Blot, Incubation

Journal: Life Science Alliance

Article Title: DOCK1 insufficiency disrupts trophoblast function and pregnancy outcomes via DUSP4-ERK pathway

doi: 10.26508/lsa.202302247

Figure Lengend Snippet: (A) Schematic illustration of the protocol for inhibiting DOCK1 during murine pregnancy. Experimental mice received vehicle control or TBOPP on GDs 6, 8, 10, and 12 through intraperitoneal injections, and were euthanatized on GDs 13.5. (B) Morphology of pregnant uteri with embryo implantation sites on GD 13.5 in mouse models after vehicle control or TBOPP treatments (n = 6 mice for each group). Scale bar: 1 cm. (C) Embryo resorption rate was calculated. (D) Western blot assays were performed to detect the levels of DUSP4, phosphorylated ERK, and HUWE1 after TBOPP treatment. (E) Quantification of DUSP4, phosphorylated ERK, and HUWE1 protein levels. (F) Representative HE-stained images of placentas and CK7 immunofluorescence of trophoblast cell invasion into the decidua after TBOPP treatment (left). Based on the HE and CK7 staining, distinct layers—Dec (decidua), Jz (junctional zone), and Lab (labyrinth)—are delineated by black lines in HE images and white lines in immunofluorescence images. Boxes on the left correspond to magnified sections on the right, with the region between the yellow and white lines indicating the trophoblast cell invasion area into the decidua. Scale bar: 500 μm. (G) The area of trophoblast cell invasion into the decidua was quantified. (H) Representative images of Ki67 immunostaining in placental tissues after TBOPP treatment (left). Percentage of Ki67-positive cells was calculated (right). Scale bar: 200 μm. In (C, E, G, H), each group, n = 6 mice (* P < 0.05; ** P < 0.01; *** P < 0.001, unpaired two-tailed t test).

Article Snippet: DUSP4 , IF(1:200) , Proteintech , #66349-1-Ig.

Techniques: Control, Western Blot, Staining, Immunofluorescence, Immunostaining, Two Tailed Test

Journal: Life Science Alliance

Article Title: DOCK1 insufficiency disrupts trophoblast function and pregnancy outcomes via DUSP4-ERK pathway

doi: 10.26508/lsa.202302247

Figure Lengend Snippet: Details of antibodies used.

Article Snippet: DUSP4 , IF(1:200) , Proteintech , #66349-1-Ig.

Techniques:

Journal: Life Science Alliance

Article Title: DOCK1 insufficiency disrupts trophoblast function and pregnancy outcomes via DUSP4-ERK pathway

doi: 10.26508/lsa.202302247

Figure Lengend Snippet: Primers used for real-time quantitative polymerase chain reaction.

Article Snippet: DUSP4 , IF(1:200) , Proteintech , #66349-1-Ig.

Techniques:

Journal: Frontiers in Microbiology

Article Title: Echinococcus granulosus sensu lato promotes osteoclast differentiation through DUSP4-MAPK signaling in osseous echinococcosis

doi: 10.3389/fmicb.2025.1558603

Figure Lengend Snippet: PSC intervention downregulates DUSP4 and upregulates the MAPK signaling pathway. (A) mRNA expression levels of DUSP2 and DUSP4 following different doses of PSC intervention. (B) Protein expression levels of DUSP4 after different doses of PSC intervention. (C) Quantification of DUSP4 protein levels shown in (B) . (D) Visualization of F-actin ring formation and DUSP4 expression in osteoclasts after different doses of PSC intervention. Green: F-actin; red: DUSP4; blue: DAPI. Magnification ×40. (E) Quantification of the F-actin ring area and the absolute and relative fluorescence intensity of DUSP4 shown in (D) . Data represent three independent experiments. ns, not significant; * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: The membrane was blocked with 5% bovine serum albumin (BSA) blocking solution (BIOTEK, China) for 2 h, and then incubated overnight at 4°C with primary antibodies: DUSP4 (1:1000; Boster, #OT17C11); GAPDH (1:2000; Boster, #BG-7) ERK (1:2,000; #4695), p-ERK (1:1,000; #4370), JNK (1:2,000; #9252), p-JNK (1:1,000; #4668), p38 (1:2,000; #8690), p-p38 (1:1,000; #4511) (Cell Signaling Technology, Inc.); MMP-9 (1:1000; #ab76003); c-Fos (1:2,000; #ab190289), TRAP (1:1,000; #ab191406), NFATc1 (4 μg/mL; #ab2796), Cathepsin K (1:2,000; #ab19027) (Abcam, Inc).

Techniques: Expressing, Fluorescence

Journal: Frontiers in Microbiology

Article Title: Echinococcus granulosus sensu lato promotes osteoclast differentiation through DUSP4-MAPK signaling in osseous echinococcosis

doi: 10.3389/fmicb.2025.1558603

Figure Lengend Snippet: Overexpression of DUSP4 affects osteoclast formation. (A) mRNA expression levels of DUSP4 in control, lentiviral-transfected null, and DUSP4-overexpression groups. (B) Protein levels of DUSP4 in the same groups. (C) Quantification of (B) . (D) Cell viability assay in the same groups. (E) TRAP staining of osteoclasts in control and DUSP4-overexpression groups after PSC intervention. Magnification ×40. (F) Quantification of the area occupied by TRAP-positive cells and the number of TRAP-positive cells shown in (E) . (G) F-actin ring staining in control and DUSP4-overexpression groups after PSC intervention green: F-actin; blue: DAPI. Magnification ×40. (H) Quantification of the F-actin ring area and fluorescence intensity shown in (G) . Data represent three independent experiments, and significance was determined using one-way ANOVA. ns, no significance; * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: The membrane was blocked with 5% bovine serum albumin (BSA) blocking solution (BIOTEK, China) for 2 h, and then incubated overnight at 4°C with primary antibodies: DUSP4 (1:1000; Boster, #OT17C11); GAPDH (1:2000; Boster, #BG-7) ERK (1:2,000; #4695), p-ERK (1:1,000; #4370), JNK (1:2,000; #9252), p-JNK (1:1,000; #4668), p38 (1:2,000; #8690), p-p38 (1:1,000; #4511) (Cell Signaling Technology, Inc.); MMP-9 (1:1000; #ab76003); c-Fos (1:2,000; #ab190289), TRAP (1:1,000; #ab191406), NFATc1 (4 μg/mL; #ab2796), Cathepsin K (1:2,000; #ab19027) (Abcam, Inc).

Techniques: Over Expression, Expressing, Control, Transfection, Viability Assay, Staining, Fluorescence

Journal: Frontiers in Microbiology

Article Title: Echinococcus granulosus sensu lato promotes osteoclast differentiation through DUSP4-MAPK signaling in osseous echinococcosis

doi: 10.3389/fmicb.2025.1558603

Figure Lengend Snippet: Effects of DUSP4 on the MAPK pathway and osteoclast-related genes. (A) Western blot analysis showing changes in MAPK signaling pathway components (p-JNK, p-ERK, and p-p38) in osteoclasts after PSC intervention in control and DUSP4-overexpression groups. (B) Quantification of MAPK pathway phosphorylation levels in osteoclasts from control and DUSP4-overexpression groups, as shown in (A) . (C) mRNA expression levels of osteoclast-related proteins (DUSP4, MMP9, NFATc1, TRAP, and CTSK) after PSC intervention in control and DUSP4-overexpression groups. (D) Western blot analysis of osteoclast-related proteins (DUSP4, MMP9, NFATc1, CTSK, and c-fos) after PSC intervention in control and overexpression groups. (E) Quantification of osteoclast-related proteins expression levels in (D) . Data represent three independent experiments, and significance was determined using one-way ANOVA. ns, no significance; * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: The membrane was blocked with 5% bovine serum albumin (BSA) blocking solution (BIOTEK, China) for 2 h, and then incubated overnight at 4°C with primary antibodies: DUSP4 (1:1000; Boster, #OT17C11); GAPDH (1:2000; Boster, #BG-7) ERK (1:2,000; #4695), p-ERK (1:1,000; #4370), JNK (1:2,000; #9252), p-JNK (1:1,000; #4668), p38 (1:2,000; #8690), p-p38 (1:1,000; #4511) (Cell Signaling Technology, Inc.); MMP-9 (1:1000; #ab76003); c-Fos (1:2,000; #ab190289), TRAP (1:1,000; #ab191406), NFATc1 (4 μg/mL; #ab2796), Cathepsin K (1:2,000; #ab19027) (Abcam, Inc).

Techniques: Western Blot, Control, Over Expression, Phospho-proteomics, Expressing

Journal: Frontiers in Microbiology

Article Title: Echinococcus granulosus sensu lato promotes osteoclast differentiation through DUSP4-MAPK signaling in osseous echinococcosis

doi: 10.3389/fmicb.2025.1558603

Figure Lengend Snippet: DUSP4 inhibits osteoclasts and attenuates bone destruction in osseous echinococcosis. (A) X-ray examination of long bones from mice treated with empty vector, lentiviral DUSP4, or left untreated in the affection of PSC after 6 months, with red rectangle indicating cyst. (B) Micro-CT images of long bones from each treatment group, with red arrows indicating bone defects. (C) HE staining of bone tissue sections from each treatment group. Magnification ×20. (D) Western blot analysis of osteoclast-related proteins (DUSP4 and CTSK) after PSC intervention in control and DUSP4-overexpression groups. (E) Quantification of osteoclast-related protein expression levels in osteoclasts from control and DUSP4-overexpression groups, as shown in (D) . Data are presented as mean ± SD from five mice per group. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: The membrane was blocked with 5% bovine serum albumin (BSA) blocking solution (BIOTEK, China) for 2 h, and then incubated overnight at 4°C with primary antibodies: DUSP4 (1:1000; Boster, #OT17C11); GAPDH (1:2000; Boster, #BG-7) ERK (1:2,000; #4695), p-ERK (1:1,000; #4370), JNK (1:2,000; #9252), p-JNK (1:1,000; #4668), p38 (1:2,000; #8690), p-p38 (1:1,000; #4511) (Cell Signaling Technology, Inc.); MMP-9 (1:1000; #ab76003); c-Fos (1:2,000; #ab190289), TRAP (1:1,000; #ab191406), NFATc1 (4 μg/mL; #ab2796), Cathepsin K (1:2,000; #ab19027) (Abcam, Inc).

Techniques: Plasmid Preparation, Micro-CT, Staining, Western Blot, Control, Over Expression, Expressing

Journal: Frontiers in Microbiology

Article Title: Echinococcus granulosus sensu lato promotes osteoclast differentiation through DUSP4-MAPK signaling in osseous echinococcosis

doi: 10.3389/fmicb.2025.1558603

Figure Lengend Snippet:

Article Snippet: The membrane was blocked with 5% bovine serum albumin (BSA) blocking solution (BIOTEK, China) for 2 h, and then incubated overnight at 4°C with primary antibodies: DUSP4 (1:1000; Boster, #OT17C11); GAPDH (1:2000; Boster, #BG-7) ERK (1:2,000; #4695), p-ERK (1:1,000; #4370), JNK (1:2,000; #9252), p-JNK (1:1,000; #4668), p38 (1:2,000; #8690), p-p38 (1:1,000; #4511) (Cell Signaling Technology, Inc.); MMP-9 (1:1000; #ab76003); c-Fos (1:2,000; #ab190289), TRAP (1:1,000; #ab191406), NFATc1 (4 μg/mL; #ab2796), Cathepsin K (1:2,000; #ab19027) (Abcam, Inc).

Techniques:

Journal: International Journal of Oncology

Article Title: Microarray phosphatome profiling of breast cancer patients unveils a complex phosphatase regulatory role of the MAPK and PI3K pathways in estrogen receptor-negative breast cancers

doi: 10.3892/ijo.2014.2648

Figure Lengend Snippet: Phosphatases differentially expressed in clinical ER − ERBB2 + versus triple-negative (TN) BC patients in this series (Agilent platform) and in GSE20194 (Affymetrix platform) (both FDR q-value <0.05).

Article Snippet: The antibodies used were the rabbit polyclonal antibodies specific against the dual phosphorylated form of ERK1/2 (Thr202/Tyr204) (#4370, Cell Signaling, Beverly, MA, USA) at a dilution of 1:200, the polyclonal DUSP4 (MKP-2) antibody (NBP1-19592, Novus Biologicals, Littleton, CO, USA) at a dilution of 1:100, and a goat polyclonal anti-DUSP6 antibody (MKP-3) (sc-8599, Santa Cruz Biotechnology Lab Inc., Santa Cruz, CA, USA) at a dilution of 1:100, in the last case the anti-goat IG (HRP) (NB710-H, Novus Biologicals) was used as a secondary antibody at a dilution of 1:400.

Techniques:

Journal: International Journal of Oncology

Article Title: Microarray phosphatome profiling of breast cancer patients unveils a complex phosphatase regulatory role of the MAPK and PI3K pathways in estrogen receptor-negative breast cancers

doi: 10.3892/ijo.2014.2648

Figure Lengend Snippet: Phosphatases differentially expressed between the molecular ER − ERBB2 + and the basal-like enriched BC in this series (FDR q-value <0.05) and in the NKI series (FDR q-value ≤0.01).

Article Snippet: The antibodies used were the rabbit polyclonal antibodies specific against the dual phosphorylated form of ERK1/2 (Thr202/Tyr204) (#4370, Cell Signaling, Beverly, MA, USA) at a dilution of 1:200, the polyclonal DUSP4 (MKP-2) antibody (NBP1-19592, Novus Biologicals, Littleton, CO, USA) at a dilution of 1:100, and a goat polyclonal anti-DUSP6 antibody (MKP-3) (sc-8599, Santa Cruz Biotechnology Lab Inc., Santa Cruz, CA, USA) at a dilution of 1:100, in the last case the anti-goat IG (HRP) (NB710-H, Novus Biologicals) was used as a secondary antibody at a dilution of 1:400.

Techniques:

Journal: International Journal of Oncology

Article Title: Microarray phosphatome profiling of breast cancer patients unveils a complex phosphatase regulatory role of the MAPK and PI3K pathways in estrogen receptor-negative breast cancers

doi: 10.3892/ijo.2014.2648

Figure Lengend Snippet: Phosphatases differentially expressed between ER + and ER − BC in common among GSE7390, GSE20194 and GSE2034 (FDR q-value ≤0.05).

Article Snippet: The antibodies used were the rabbit polyclonal antibodies specific against the dual phosphorylated form of ERK1/2 (Thr202/Tyr204) (#4370, Cell Signaling, Beverly, MA, USA) at a dilution of 1:200, the polyclonal DUSP4 (MKP-2) antibody (NBP1-19592, Novus Biologicals, Littleton, CO, USA) at a dilution of 1:100, and a goat polyclonal anti-DUSP6 antibody (MKP-3) (sc-8599, Santa Cruz Biotechnology Lab Inc., Santa Cruz, CA, USA) at a dilution of 1:100, in the last case the anti-goat IG (HRP) (NB710-H, Novus Biologicals) was used as a secondary antibody at a dilution of 1:400.

Techniques:

Journal: International Journal of Oncology

Article Title: Microarray phosphatome profiling of breast cancer patients unveils a complex phosphatase regulatory role of the MAPK and PI3K pathways in estrogen receptor-negative breast cancers

doi: 10.3892/ijo.2014.2648

Figure Lengend Snippet: Co-expression network analysis from the GeneMania server using DUSP4, DUSP6 and DUSP10 as query genes.

Article Snippet: The antibodies used were the rabbit polyclonal antibodies specific against the dual phosphorylated form of ERK1/2 (Thr202/Tyr204) (#4370, Cell Signaling, Beverly, MA, USA) at a dilution of 1:200, the polyclonal DUSP4 (MKP-2) antibody (NBP1-19592, Novus Biologicals, Littleton, CO, USA) at a dilution of 1:100, and a goat polyclonal anti-DUSP6 antibody (MKP-3) (sc-8599, Santa Cruz Biotechnology Lab Inc., Santa Cruz, CA, USA) at a dilution of 1:100, in the last case the anti-goat IG (HRP) (NB710-H, Novus Biologicals) was used as a secondary antibody at a dilution of 1:400.

Techniques: Expressing

Journal: International Journal of Oncology

Article Title: Microarray phosphatome profiling of breast cancer patients unveils a complex phosphatase regulatory role of the MAPK and PI3K pathways in estrogen receptor-negative breast cancers

doi: 10.3892/ijo.2014.2648

Figure Lengend Snippet: DUSP4, DUSP6 and phospo-ERK1/2 immuohistochemical percentage scores within the triple-negative breast carcinomas and the ERBB2-positive, ER and PR-negative breast carcinoma groups.

Article Snippet: The antibodies used were the rabbit polyclonal antibodies specific against the dual phosphorylated form of ERK1/2 (Thr202/Tyr204) (#4370, Cell Signaling, Beverly, MA, USA) at a dilution of 1:200, the polyclonal DUSP4 (MKP-2) antibody (NBP1-19592, Novus Biologicals, Littleton, CO, USA) at a dilution of 1:100, and a goat polyclonal anti-DUSP6 antibody (MKP-3) (sc-8599, Santa Cruz Biotechnology Lab Inc., Santa Cruz, CA, USA) at a dilution of 1:100, in the last case the anti-goat IG (HRP) (NB710-H, Novus Biologicals) was used as a secondary antibody at a dilution of 1:400.

Techniques: Expressing, Two Tailed Test

Journal: International Journal of Oncology

Article Title: Microarray phosphatome profiling of breast cancer patients unveils a complex phosphatase regulatory role of the MAPK and PI3K pathways in estrogen receptor-negative breast cancers

doi: 10.3892/ijo.2014.2648

Figure Lengend Snippet: Multiphosphatase signature comprising 58 probes (48 genes) trained in GSE2034 training set and validated in GSE7390 (both Affymetrix HGU133A platform).

Article Snippet: The antibodies used were the rabbit polyclonal antibodies specific against the dual phosphorylated form of ERK1/2 (Thr202/Tyr204) (#4370, Cell Signaling, Beverly, MA, USA) at a dilution of 1:200, the polyclonal DUSP4 (MKP-2) antibody (NBP1-19592, Novus Biologicals, Littleton, CO, USA) at a dilution of 1:100, and a goat polyclonal anti-DUSP6 antibody (MKP-3) (sc-8599, Santa Cruz Biotechnology Lab Inc., Santa Cruz, CA, USA) at a dilution of 1:100, in the last case the anti-goat IG (HRP) (NB710-H, Novus Biologicals) was used as a secondary antibody at a dilution of 1:400.

Techniques:

Journal: bioRxiv

Article Title: Role of mPFC DUSP1 in regulating the sensitivity to cocaine in adolescent cocaine-exposed mice of adulthood

doi: 10.1101/2022.08.26.505502

Figure Lengend Snippet: A , Dusp1 mRNA levels on P78. B , DUSP1 protein levels and activity on P78. C , DUSP1 activity on P78. D , DUSP1 staining in CaMKII-positive neurons. Scale bar, 400 μm/100 μm. E , Levels of MAPK-related signal proteins on P78, including p-ERK/ERK, p-ERK, ERK, p-CREB/CREB, p-CREB, CREB and BDNF. ASE-C, subthreshold-dosed administration of cocaine in adolescent saline-exposed mice; ACE-C, subthreshold-dosed administration of cocaine in adolescent cocaine-exposed mice; N.S., p > 0.05, *, p < 0.05, **, p < 0.01 vs ASE-C mice.

Article Snippet: The sections were incubated in 0.3% (v/v) Triton X-100 for 0.5 h, blocked with 5% donkey serum for 1.5 h at room temperature, and incubated overnight at 4°C with the following primary antibodies: rabbit anti-c-Fos (1:1500, RRID: AB_2247211, Cell Signalling Technology, USA), mouse anti-NeuN (1:800, RRID: AB_2298772, Millipore, USA), DUSP1 (1:100, BM4856, Boster Biotechnology, China), mouse anti-GAD67 (1:500, RRID: AB_2278725, Millipore, USA), mouse anti-PV (1:250, BM1339, Boster Biotechnology, China), mouse anti-SOM (1:50, RRID:AB_2271061, Santa Cruz, USA), mouse anti-CaMKII α (1:100, SC-13141, Santa Cruz, USA) and mouse anti-CaMKII α (1:200, RRID: AB_2721906, Cell Signalling Technology, USA), followed by the corresponding fluorophore-conjugated secondary antibodies for 1.5 h at room temperature.

Techniques: Activity Assay, Staining, Saline

Journal: bioRxiv

Article Title: Role of mPFC DUSP1 in regulating the sensitivity to cocaine in adolescent cocaine-exposed mice of adulthood

doi: 10.1101/2022.08.26.505502

Figure Lengend Snippet: A , DUSP1 protein levels on P74. B , DUSP1 activity on P74. ASE, adolescent salineexposed mice; ACE, adolescent cocaine-exposed mice; N.S., p > 0.05 vs ASE mice.

Article Snippet: The sections were incubated in 0.3% (v/v) Triton X-100 for 0.5 h, blocked with 5% donkey serum for 1.5 h at room temperature, and incubated overnight at 4°C with the following primary antibodies: rabbit anti-c-Fos (1:1500, RRID: AB_2247211, Cell Signalling Technology, USA), mouse anti-NeuN (1:800, RRID: AB_2298772, Millipore, USA), DUSP1 (1:100, BM4856, Boster Biotechnology, China), mouse anti-GAD67 (1:500, RRID: AB_2278725, Millipore, USA), mouse anti-PV (1:250, BM1339, Boster Biotechnology, China), mouse anti-SOM (1:50, RRID:AB_2271061, Santa Cruz, USA), mouse anti-CaMKII α (1:100, SC-13141, Santa Cruz, USA) and mouse anti-CaMKII α (1:200, RRID: AB_2721906, Cell Signalling Technology, USA), followed by the corresponding fluorophore-conjugated secondary antibodies for 1.5 h at room temperature.

Techniques: Activity Assay

Journal: bioRxiv

Article Title: Role of mPFC DUSP1 in regulating the sensitivity to cocaine in adolescent cocaine-exposed mice of adulthood

doi: 10.1101/2022.08.26.505502

Figure Lengend Snippet: A , Experimental design and timeline. B , Schematic diagram of viral injection. Scale bar, 400 μm. C , Percentage of viral transfected neurons in CaMKII-positive neurons of mPFC. Scale bar, 200 μm/100 μm. D , Dusp1 mRNA levels on P78. E , DUSP1 protein levels on P78. F , DUSP1 activity on P78. G , CPP score. H , ΔCPP score. I , Heatmap of spent duration by mice in CPP apparatus. J , C-Fos-stained value in virus-transfected neurons. Scale bar, 200 μm/100 μm. K , Dendritic spine density of pyramidal neurons in mPFC. Scale bar, 10 μm. L , Levels of MAPK-related signal proteins, including p-ERK/ERK, p-ERK, ERK, p-CREB/CREB, p-CREB, CREB and BDNF. ACE-CCtrl, ACE-C mice injected with Ctrl virus; ACE-COE, ACE-C mice with overexpression virus of DUSP1; N.S., p > 0.05, *, p < 0.05, **, p < 0.01 vs ACE-CCtrl mice.

Article Snippet: The sections were incubated in 0.3% (v/v) Triton X-100 for 0.5 h, blocked with 5% donkey serum for 1.5 h at room temperature, and incubated overnight at 4°C with the following primary antibodies: rabbit anti-c-Fos (1:1500, RRID: AB_2247211, Cell Signalling Technology, USA), mouse anti-NeuN (1:800, RRID: AB_2298772, Millipore, USA), DUSP1 (1:100, BM4856, Boster Biotechnology, China), mouse anti-GAD67 (1:500, RRID: AB_2278725, Millipore, USA), mouse anti-PV (1:250, BM1339, Boster Biotechnology, China), mouse anti-SOM (1:50, RRID:AB_2271061, Santa Cruz, USA), mouse anti-CaMKII α (1:100, SC-13141, Santa Cruz, USA) and mouse anti-CaMKII α (1:200, RRID: AB_2721906, Cell Signalling Technology, USA), followed by the corresponding fluorophore-conjugated secondary antibodies for 1.5 h at room temperature.

Techniques: Injection, Transfection, Activity Assay, Staining, Virus, Over Expression

Journal: bioRxiv

Article Title: Role of mPFC DUSP1 in regulating the sensitivity to cocaine in adolescent cocaine-exposed mice of adulthood

doi: 10.1101/2022.08.26.505502

Figure Lengend Snippet: A , Experimental design and timeline. B , Schematic diagram of viral injection. Scale bar, 400 μm. C , Percentage of viral transfected neurons in CaMKII-positive neurons of mPFC. Scale bar, 200 μm/100 μm. D , Dusp1 mRNA levels on P78. E , DUSP1 protein levels on P78. F , DUSP1 activity on P78. G , CPP score. H , ΔCPP score. I , Heatmap of spent duration by mice in CPP apparatus. J , C-Fos-stained value in virus-transfected neurons. Scale bar, 200 μm/100 μm. K , Dendritic spine density of pyramidal neurons in mPFC. Scale bar, 10 μm. L , Levels of MAPK-related signal proteins, including p-ERK/ERK, p-ERK, ERK, p-CREB/CREB, p-CREB, CREB and BDNF. ACE-CCtrl, ACE-C mice injected with Ctrl virus; ACE-CKD, ACE-C mice with knockdown virus of DUSP1; N.S., p > 0.05, *, p < 0.05, **, p < 0.01 vs ACE-CCtrl mice.

Article Snippet: The sections were incubated in 0.3% (v/v) Triton X-100 for 0.5 h, blocked with 5% donkey serum for 1.5 h at room temperature, and incubated overnight at 4°C with the following primary antibodies: rabbit anti-c-Fos (1:1500, RRID: AB_2247211, Cell Signalling Technology, USA), mouse anti-NeuN (1:800, RRID: AB_2298772, Millipore, USA), DUSP1 (1:100, BM4856, Boster Biotechnology, China), mouse anti-GAD67 (1:500, RRID: AB_2278725, Millipore, USA), mouse anti-PV (1:250, BM1339, Boster Biotechnology, China), mouse anti-SOM (1:50, RRID:AB_2271061, Santa Cruz, USA), mouse anti-CaMKII α (1:100, SC-13141, Santa Cruz, USA) and mouse anti-CaMKII α (1:200, RRID: AB_2721906, Cell Signalling Technology, USA), followed by the corresponding fluorophore-conjugated secondary antibodies for 1.5 h at room temperature.

Techniques: Injection, Transfection, Activity Assay, Staining, Virus, Knockdown

Journal: bioRxiv

Article Title: Role of mPFC DUSP1 in regulating the sensitivity to cocaine in adolescent cocaine-exposed mice of adulthood

doi: 10.1101/2022.08.26.505502

Figure Lengend Snippet: In adulthood, mice with cocaine-exposed experience during adolescent period (ACE-C mice) exhibits higher sensitivity to cocaine, accompanied by more activated mPFC and lower levels of DUSP1 protein and activity of mPFC in response to subthreshold-dosed administration of cocaine. Specific overexpression of DUSP1 on mPFC glutamatergic neurons efficiently blocks cocaine-preferred behaviors and reduces mPFC activity, while knockdown of DUSP1 maintains cocaine-preferred behaviors and increases mPFC activity in ACE-C mice. It might be through regulating MAPK signal molecules such as ERK1/2 that DUSP1 regulates the enhanced mPFC activation and sensitivity of ACE mice to cocaine.

Article Snippet: The sections were incubated in 0.3% (v/v) Triton X-100 for 0.5 h, blocked with 5% donkey serum for 1.5 h at room temperature, and incubated overnight at 4°C with the following primary antibodies: rabbit anti-c-Fos (1:1500, RRID: AB_2247211, Cell Signalling Technology, USA), mouse anti-NeuN (1:800, RRID: AB_2298772, Millipore, USA), DUSP1 (1:100, BM4856, Boster Biotechnology, China), mouse anti-GAD67 (1:500, RRID: AB_2278725, Millipore, USA), mouse anti-PV (1:250, BM1339, Boster Biotechnology, China), mouse anti-SOM (1:50, RRID:AB_2271061, Santa Cruz, USA), mouse anti-CaMKII α (1:100, SC-13141, Santa Cruz, USA) and mouse anti-CaMKII α (1:200, RRID: AB_2721906, Cell Signalling Technology, USA), followed by the corresponding fluorophore-conjugated secondary antibodies for 1.5 h at room temperature.

Techniques: Activity Assay, Over Expression, Knockdown, Activation Assay

Journal: Breast Cancer Research and Treatment

Article Title: Analysis of phosphatases in ER-negative breast cancers identifies DUSP4 as a critical regulator of growth and invasion

doi: 10.1007/s10549-016-3892-y

Figure Lengend Snippet: Genomic alteration and expression of phosphatases. a Percent of all tumors with any deletion among the top five underexpressed genes in the TCGA dataset. b Homozygous deletion of selected phosphatases in TCGA. c All somatic alterations of DUSP4 gene represented by the cBio Oncoprint. d Differential expression of DUSP4 in two breast cancer datasets

Article Snippet: The DUSP4 antibody was purchased from BD Transduction Laboratories (Lexington, KY, USA).

Techniques: Expressing

Journal: Breast Cancer Research and Treatment

Article Title: Analysis of phosphatases in ER-negative breast cancers identifies DUSP4 as a critical regulator of growth and invasion

doi: 10.1007/s10549-016-3892-y

Figure Lengend Snippet: Induced expression of DUSP4 inhibits ER-negative but not ER-positive growth in vitro. a Western blot analysis of doxycycline-induced expression of DUSP4 in SUM159, MDA-MB-231, and MCF7 breast cancer cells. b – d Proliferation analysis of breast cancer cell lines upon expression of DUSP4 . e – g Anchorage-independent colony formation assay of breast cancer cell lines upon expression of DUSP4

Article Snippet: The DUSP4 antibody was purchased from BD Transduction Laboratories (Lexington, KY, USA).

Techniques: Expressing, In Vitro, Western Blot, Colony Assay

Journal: Breast Cancer Research and Treatment

Article Title: Analysis of phosphatases in ER-negative breast cancers identifies DUSP4 as a critical regulator of growth and invasion

doi: 10.1007/s10549-016-3892-y

Figure Lengend Snippet: Induction of DUSP4 expression inhibits the growth of ER-negative breast cancer cells in vivo. a Induced expression of DUSP4 does not inhibit in vivo xenograft growth of MDA-MB-231 vector clone ±Dox; b calculated slope from MDA-MB-231 vector clone xenograft. c MDA-MB-231-DUSP4 clone ±Dox growth curves; d calculated slope from MDA-MB-231-DUSP4 xenograft. e Induced expression of DUSP4 does not inhibit in vivo xenograft growth of SUM 159-Vector clone ±Dox growth curves; f calculated slope from SUM 159-vector ±Dox, g SUM 159-DUSP4 clone ±Dox growth curves; h calculated slope from SUM 159-DUSP4 clone xenograft. t -test p values are indicated

Article Snippet: The DUSP4 antibody was purchased from BD Transduction Laboratories (Lexington, KY, USA).

Techniques: Expressing, In Vivo, Plasmid Preparation

Journal: Breast Cancer Research and Treatment

Article Title: Analysis of phosphatases in ER-negative breast cancers identifies DUSP4 as a critical regulator of growth and invasion

doi: 10.1007/s10549-016-3892-y

Figure Lengend Snippet: Increased DUSP4 expression causes reduced proliferation, invasion, and a G1 cell cycle block. a Representative Ki67 staining of MDA-MB-231 xenograft sections. b Quantitation of Ki67; c representative pictures of Boyden chamber assay for migration for the cells before and after DUSP4 induction. d Relative migration in MDA-MB-231; e representative pictures of Boyden chamber assay for invasion for the cells before and after DUSP4 induction. f Relative invasion in MDA-MB-231. g Cell cycle changes after DUSP4 induction. t -test p values are indicated

Article Snippet: The DUSP4 antibody was purchased from BD Transduction Laboratories (Lexington, KY, USA).

Techniques: Expressing, Blocking Assay, Staining, Quantitation Assay, Boyden Chamber Assay, Migration

Journal: Breast Cancer Research and Treatment

Article Title: Analysis of phosphatases in ER-negative breast cancers identifies DUSP4 as a critical regulator of growth and invasion

doi: 10.1007/s10549-016-3892-y

Figure Lengend Snippet: RPPA analysis identifies signaling pathways altered upon induced expression of DUSP4. a RPPA analysis was performed on SUM159 cells with and without induction of DUSP4 (–Dox = No DUSP4 over expression and +Dox = DUSP4 overexpression). Selective markedly altered proteins are indicated after DUSP4 expression. The median expression values for each protein in cells treated with Dox was subtracted from the protein expression value in cells treated with vehicle to obtain a difference value for each protein studied. Those proteins that showed the greatest change (increased or decreased) are shown in Supplementary Table 2. b Changes in MAP Kinase group: p-ERK1/2, p-p38, and p-JNK1/2. c Changes in the other signaling pathways: p-Rb, p-p65 (S536), and p-AMPK (T172)

Article Snippet: The DUSP4 antibody was purchased from BD Transduction Laboratories (Lexington, KY, USA).

Techniques: Expressing, Over Expression

Journal: Breast Cancer Research and Treatment

Article Title: Analysis of phosphatases in ER-negative breast cancers identifies DUSP4 as a critical regulator of growth and invasion

doi: 10.1007/s10549-016-3892-y

Figure Lengend Snippet: Differential expression of overexpressed phosphatases in ER-negative versus ER-positive breast cancer

Article Snippet: The DUSP4 antibody was purchased from BD Transduction Laboratories (Lexington, KY, USA).

Techniques: Expressing

Journal: Stem Cells International

Article Title: Exosomal miR-423-5p Derived from Cerebrospinal Fluid Pulsation Stress-Stimulated Osteoblasts Improves Angiogenesis of Endothelial Cells via DUSP8/ERK1/2 Signaling Pathway

doi: 10.1155/2024/5512423

Figure Lengend Snippet: (a) bEnd.3 cells cocultured with OBs transfected with Cy3-labeled miR-423-5p mimic (red) in a transwell (0.4 μ m) plate; (b) representative images of OBs transfected with Cy3-miR-423-5p mimic; (c) the effects of GW4869 on exosome-dependent miRNA delivery from OBs into bEnd.3 cells; (d) the binging site for miRNA in DUSP8 mRNA; (e) a luciferase report assay; (f and g) western blotting. Data are represented as mean ± SD; n = 3. ∗ P < 0.05 vs. the Mimic, ∗∗ P < 0.01 vs. the Dusp8-WT NC, ∗∗∗ P < 0.001 vs. the Control, # P < 0.05 vs. the Inhibitor, ## P < 0.01 vs. the +OBs, ns P > 0.05 vs. Dusp8-WU NC.

Article Snippet: After being blocked with 5% nonfat dried milk, the membranes were incubated with primary antibodies for β -actin (CST, USA, Cat 4967S), CD63 (ABclonal, China, Cat A19023), CD81 (ABclonal, China, Cat A22528), DUSP8 (ABclonal, China, Cat A17990), ERK1/2 (CST, USA, Cat 4695S), pERK1/2 (CST, USA, Cat 4376S) at 4°C overnight.

Techniques: Transfection, Labeling, Luciferase, Western Blot, Control

Journal: Stem Cells International

Article Title: Exosomal miR-423-5p Derived from Cerebrospinal Fluid Pulsation Stress-Stimulated Osteoblasts Improves Angiogenesis of Endothelial Cells via DUSP8/ERK1/2 Signaling Pathway

doi: 10.1155/2024/5512423

Figure Lengend Snippet: Exosomal miR-423-5p regulates angiogenesis in vivo: (a) image of Matrigel plugs after resection at 14 days; (b and c) western blot analysis of DUSP8 expression levels; (d, f, and g) immunohistochemical images analysis of p-ERK and VEGF-positive from Matrigel plugs; (e and h) images of CD31 detected by immunofluorescence staining in groups. ∗ P < 0.05 vs. the NC-Exos, ∗∗ P < 0.01 vs. the NC-Exos, ∗∗∗ P < 0.001 vs. the NC-Exos, ∗∗∗∗ P < 0.0001 vs. the NC-Exos.

Article Snippet: After being blocked with 5% nonfat dried milk, the membranes were incubated with primary antibodies for β -actin (CST, USA, Cat 4967S), CD63 (ABclonal, China, Cat A19023), CD81 (ABclonal, China, Cat A22528), DUSP8 (ABclonal, China, Cat A17990), ERK1/2 (CST, USA, Cat 4695S), pERK1/2 (CST, USA, Cat 4376S) at 4°C overnight.

Techniques: In Vivo, Western Blot, Expressing, Immunohistochemical staining, Immunofluorescence, Staining

Journal: Stem Cells International

Article Title: Exosomal miR-423-5p Derived from Cerebrospinal Fluid Pulsation Stress-Stimulated Osteoblasts Improves Angiogenesis of Endothelial Cells via DUSP8/ERK1/2 Signaling Pathway

doi: 10.1155/2024/5512423

Figure Lengend Snippet: Model of exosomal miR-423-5p derived from CSFP-induced OBs in promoting angiogenesis of ECs. CSFP induces the secretion of OBs-derived exosomal miR-423-5p. OBs-derived exosomal miR-423-5p can directly target DUSP8 and then regulate the ERK1/2 signaling pathway to promote the angiogenesis of ECs.

Article Snippet: After being blocked with 5% nonfat dried milk, the membranes were incubated with primary antibodies for β -actin (CST, USA, Cat 4967S), CD63 (ABclonal, China, Cat A19023), CD81 (ABclonal, China, Cat A22528), DUSP8 (ABclonal, China, Cat A17990), ERK1/2 (CST, USA, Cat 4695S), pERK1/2 (CST, USA, Cat 4376S) at 4°C overnight.

Techniques: Derivative Assay