Journal: Scientific Reports

Article Title: PTPRK promotes resiniferatoxin-induced postherpetic neuralgia via activating DUSP1/p38 MAPK signaling pathway in dorsal root ganglia

doi: 10.1038/s41598-025-24233-y

Figure Lengend Snippet: PTPRK activated the DUSP1/p38 MAPK signaling pathway in PHN. A , Western blot was performed to analyze the protein levels of DUSP1, p-p38 MAPK, and p38 MAPK in DRG tissues of each group; B , The protein levels of PTPRK, DUSP1, p-p38 MAPK, and p38 MAPK in Lv-NC and Lv-PTPRK groups were detected via Western blot. ( N = 5).

Article Snippet: The used antibodies are shown here: PTPRK (28151-1-AP; 1/1000; Proteintech Biotech Co. Ltd., Wuhan, China), DUSP1 (HY- P80645 ; 1/1000; MedChem Express), p38 (HY- P80996 ; 1/1000; MedChem Express), phosphorylated p38 (p-p38; 3485; 1/1000; Cell Signaling Technology, Danvers, MA, USA), GAPDH (10494-1-AP; 1/5000; Proteintech), and goat-anti-rabbit IgG (SA00001-2; 1/10,000; Proteintech).

Techniques: Western Blot

Journal: Scientific Reports

Article Title: PTPRK promotes resiniferatoxin-induced postherpetic neuralgia via activating DUSP1/p38 MAPK signaling pathway in dorsal root ganglia

doi: 10.1038/s41598-025-24233-y

Figure Lengend Snippet: p38 MAPK signaling pathway activation promoted mechanical allodynia and inflammation in rat DRG tissues. A , Paw withdrawal thresholds and B , paw withdrawal latency in each group were quantified; ELISA was performed to assess the concentrations of C , TNF-α, D , IL-1β, and E , IL-4 in DRG tissues of each group; RT-qPCR was performed to analyze the mRNA levels of F , TNF-α, G , IL-1β, H , IL-4, I , BDNF, J , Nav1.3, K , Nav1.7, and L , TRPV1 in DRG tissues of each group. ( N = 5; ** p < 0.01 vs. the RTX-PHN + Lv-shNC group; # p < 0.05, ## p < 0.01 vs. the RTX-PHN + Lv-shPTPRK group).

Article Snippet: The used antibodies are shown here: PTPRK (28151-1-AP; 1/1000; Proteintech Biotech Co. Ltd., Wuhan, China), DUSP1 (HY- P80645 ; 1/1000; MedChem Express), p38 (HY- P80996 ; 1/1000; MedChem Express), phosphorylated p38 (p-p38; 3485; 1/1000; Cell Signaling Technology, Danvers, MA, USA), GAPDH (10494-1-AP; 1/5000; Proteintech), and goat-anti-rabbit IgG (SA00001-2; 1/10,000; Proteintech).

Techniques: Activation Assay, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: PTPRK promotes resiniferatoxin-induced postherpetic neuralgia via activating DUSP1/p38 MAPK signaling pathway in dorsal root ganglia

doi: 10.1038/s41598-025-24233-y

Figure Lengend Snippet: PTPRK overexpression promoted inflammation via activating DUSP1/p38 MAPK signaling pathway in rat DRG cells. A , PTPRK expression was analyzed by RT-qPCR after empty and PTPRK overexpression vectors transfection; B , The protein levels of PTPRK, DUSP1, p-p38 MAPK, and p38 MAPK in each group were detected though Western blot; ELISA was performed to assess the concentrations of C , TNF-α, D , IL-1β, and E , IL-4 in DRG cells of each group; RT-qPCR was performed to analyze the mRNA levels of F , TNF-α, G , IL-1β, H , IL-4, I , BDNF, J , Nav1.3, K , Nav1.7, and L , TRPV1 in DRG cells of each group. ( N = 3; ** p < 0.01 vs. the vector group; ## p < 0.01 vs. the PTPRK group).

Article Snippet: The used antibodies are shown here: PTPRK (28151-1-AP; 1/1000; Proteintech Biotech Co. Ltd., Wuhan, China), DUSP1 (HY- P80645 ; 1/1000; MedChem Express), p38 (HY- P80996 ; 1/1000; MedChem Express), phosphorylated p38 (p-p38; 3485; 1/1000; Cell Signaling Technology, Danvers, MA, USA), GAPDH (10494-1-AP; 1/5000; Proteintech), and goat-anti-rabbit IgG (SA00001-2; 1/10,000; Proteintech).

Techniques: Over Expression, Expressing, Quantitative RT-PCR, Transfection, Western Blot, Enzyme-linked Immunosorbent Assay, Plasmid Preparation

Journal: bioRxiv

Article Title: The Cardioprotective Effect of Ischemic Postconditioning is Mediated by Inhibiting RAP2C-MAP4K4 Pathway

doi: 10.1101/2024.12.04.626922

Figure Lengend Snippet: a. Protein interactions based on the analysis of the BioGRID and STRING databases. b. Immunohistochemical analysis of MAP4K4 expression in rat myocardial tissue exposed to sham, I/R and I/R-postC in rat myocardial tissue (scale bar = 25 µm). c. MAP4K4 expression in rat cardiomyocytes exposed to sham conditions, ischemia/reperfusion (I/R), or I/R + PostC. d. MAP4K4 expression in rat cardiomyocytes exposed to normoxia (N), hypoxia/reoxygenation (H/R), or H/R + PostC. e. MAP4K4 expression in cardiomyocytes transfected with si-RAP2C or si-NC and exposed to H/R or H/R + PostC. f. MAP4K4 expression in cardiomyocytes transduced with Ad-RAP2C or Ad-null and exposed to H/R or H/R + PostC. g. Co-immunoprecipitation assays using cardiomyocyte lysates and RAP2C antibody under normoxia, H/R, and H/R + PostC. MAP4K4 and RAP2C expression in input and immunoprecipitated samples was evaluated by western blotting. h. Colocalization of MAP4K4 and RAP2C by immunofluorescence double staining (scale bar = 5/10 μm). The degree of colocalization is shown in scatter plots and was calculated using Pearson’s correlation coefficient. Data are means and standard deviations of three biological replicates. p values were calculated by two-tailed Student’s t- test. one-way ANOVA with Tukey’s post hoc test was used for multiple group comparisons. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Myocardial tissue was fixed in 4% paraformaldehyde for 10 min and permeabilized in Triton X-100 1% for 20 min. Endogenous peroxidase activity was blocked using 5% BSA for 1 h. Tissues were incubated with primary antibodies against RAP2C (orb474405, Biorbyt, Cambridge, UK) and MAP4K4 (sc-100445, Santa Cruz) overnight at 4℃, followed by incubation with CoraLite594-conjugated goat anti-mouse antibody (SA00013-3, Proteintech) and CoraLite488-conjugated goat anti-rabbit antibody (SA00013-2, Proteintech) at room temperature for 1 h. Nuclei were stained with DAPI.

Techniques: Immunohistochemical staining, Expressing, Transfection, Transduction, Immunoprecipitation, Western Blot, Immunofluorescence, Double Staining, Two Tailed Test

Journal: bioRxiv

Article Title: The Cardioprotective Effect of Ischemic Postconditioning is Mediated by Inhibiting RAP2C-MAP4K4 Pathway

doi: 10.1101/2024.12.04.626922

Figure Lengend Snippet: a. Western blot analysis of the efficiency of siRNA-mediated silencing of MAP4K4 expression. b. p-ERK, p-JNK, and p-P38 expression in cardiomyocytes transfected with si-MAP4K4 or si-NC and exposed to normoxia (N), H/R, or H/R + PostC. c. Number of TUNEL-positive cells in different groups (×10) (scale bar = 200 µm). d. Flow cytometry analysis of total apoptosis rates in different groups. e. Western blot analysis of the expression of cleaved caspase-3 and −9 and the Bax/Bcl-2 ratio. Data are means and standard deviations of three biological replicates. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Myocardial tissue was fixed in 4% paraformaldehyde for 10 min and permeabilized in Triton X-100 1% for 20 min. Endogenous peroxidase activity was blocked using 5% BSA for 1 h. Tissues were incubated with primary antibodies against RAP2C (orb474405, Biorbyt, Cambridge, UK) and MAP4K4 (sc-100445, Santa Cruz) overnight at 4℃, followed by incubation with CoraLite594-conjugated goat anti-mouse antibody (SA00013-3, Proteintech) and CoraLite488-conjugated goat anti-rabbit antibody (SA00013-2, Proteintech) at room temperature for 1 h. Nuclei were stained with DAPI.

Techniques: Western Blot, Expressing, Transfection, TUNEL Assay, Flow Cytometry

Journal: bioRxiv

Article Title: The Cardioprotective Effect of Ischemic Postconditioning is Mediated by Inhibiting RAP2C-MAP4K4 Pathway

doi: 10.1101/2024.12.04.626922

Figure Lengend Snippet: a. Western blot analysis of RAP2C and MAP4K4 expression in cells transfected with Ad-RAP2C or Ad-RAP2C + si-MAP4K4 and exposed to hypoxia/reoxygenation (H/R) and ischemic postconditioning (PostC). b. Protein expression of p-ERK, p-JNK, and p-P38 in different groups. c. Number of TUNEL-positive cells in different groups (×10) (scale bar = 100 µm). d. Flow cytometry analysis of the rate of apoptosis. e. Western blot analysis of the Bax/Bcl-2 ratio and the expression of cleaved caspase-3 and −9 in cardiomyocytes. Data are means and standard deviations of three biological replicates. p values were calculated by two-tailed Student’s t- test. one-way ANOVA with Tukey’s post hoc test was used for multiple group comparisons. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Myocardial tissue was fixed in 4% paraformaldehyde for 10 min and permeabilized in Triton X-100 1% for 20 min. Endogenous peroxidase activity was blocked using 5% BSA for 1 h. Tissues were incubated with primary antibodies against RAP2C (orb474405, Biorbyt, Cambridge, UK) and MAP4K4 (sc-100445, Santa Cruz) overnight at 4℃, followed by incubation with CoraLite594-conjugated goat anti-mouse antibody (SA00013-3, Proteintech) and CoraLite488-conjugated goat anti-rabbit antibody (SA00013-2, Proteintech) at room temperature for 1 h. Nuclei were stained with DAPI.

Techniques: Western Blot, Expressing, Transfection, TUNEL Assay, Flow Cytometry, Two Tailed Test