ptpn1  (Proteintech)

Journal: Experimental & Molecular Medicine

Article Title: Megakaryocytic TGFβ1 orchestrates osteogenesis of LepR + SSCs to alleviate radiation-induced bone loss

doi: 10.1038/s12276-025-01612-z

Figure Lengend Snippet: a Serum zinc concentration in mice 4 weeks after irradiation with administration of TPO or vehicle ( n = 6 per group). b Representative fluozin-3 images and quantitative analysis of LepR + SSCs, with or without MKs, after irradiation ( n = 6 per group). c Representative immunostaining images of Slc39a14 (green) in LepR + SSCs, with or without MKs, after irradiation ( n = 6 per group). Scale bar, 100 µm. d KEGG enrichment analysis of upregulated pathways in LepR + SSCs after irradiation. e GO enrichment analysis of downregulated functions in LepR + SSCs after co-culture with MKs. The top ten enriched GO terms ( P < 0.05) are shown. f Western blotting analysis of the expression of Slc39a14, PTP1B, p-eIF2α, ATF4 and CHOP in LepR + SSCs after co-culture with MKs ( n = 3 per group). g Representative immunostaining images of CHOP (green) in LepR + SSCs, with or without MKs, after irradiation ( n = 6 per group). Scale bar, 100 µm. h Transmission electron microscopy images of LepR + SSCs after irradiation co-culture with MKs ( n = 3 per group). Data on graphs are shown as mean ± SD. One-way ANOVA was used to analyze the data in a – d and g . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.

Article Snippet: Briefly, the bone sections were incubated with primary antibodies against mouse osteocalcin (A20800, ABclonal), Ki67 (AF7617, R&D), PTP1B (bs-55182R, Bioss), Slc39a14 (A10413, ABclonal), leptin receptor (bs-0410R, Bioss), CHOP ( A21902 , ABclonal), F4/80 (30325, CST), TGFβ1 (ab313729, abcam), vWF (bsm-52775R, Bioss), osterix (ab209484, abcam) and Smad2 (A7699, ABclonal) overnight at 4 °C and incubated with secondary antibodies for 1 h at 37 °C.

Techniques: Concentration Assay, Irradiation, Immunostaining, Co-Culture Assay, Western Blot, Expressing, Transmission Assay, Electron Microscopy

Journal: Experimental & Molecular Medicine

Article Title: Megakaryocytic TGFβ1 orchestrates osteogenesis of LepR + SSCs to alleviate radiation-induced bone loss

doi: 10.1038/s12276-025-01612-z

Figure Lengend Snippet: a Representative immunostaining images of LepR (red) and PTP1B (green) in the BM of irradiated mice ( n = 6 mice per group). Scale bar, 100 µm. b Representative immunostaining images of LepR (red) and PTP1B (green) in the BM of MK deleted mice and their littermate controls after irradiation ( n = 6 mice per group). Scale bar, 100 µm. c Representative immunostaining images of PTP1B in LepR + cells, with or without, MKs from the BM of TGFβ1 MKΔ/Δ and TGFβ1 fl/fl mice after irradiation ( n = 6 per group). Scale bar, 100 µm. d Western blotting analysis of the expression of PTP1B and p-Stat3 in LepR + SSCs after co-culture with MKs ( n = 3 per group), inh = inhibitor. e Western blotting analysis of the expression of PTP1B and p-Stat3 in LepR + SSCs after co-culture with MKs from the BM of TGFβ1 MKΔ/Δ and TGFβ1 fl/fl mice ( n = 3 per group). Data on graphs are shown as mean ± SD. One-way ANOVA was used to analyze the data in c and d . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.

Article Snippet: Briefly, the bone sections were incubated with primary antibodies against mouse osteocalcin (A20800, ABclonal), Ki67 (AF7617, R&D), PTP1B (bs-55182R, Bioss), Slc39a14 (A10413, ABclonal), leptin receptor (bs-0410R, Bioss), CHOP ( A21902 , ABclonal), F4/80 (30325, CST), TGFβ1 (ab313729, abcam), vWF (bsm-52775R, Bioss), osterix (ab209484, abcam) and Smad2 (A7699, ABclonal) overnight at 4 °C and incubated with secondary antibodies for 1 h at 37 °C.

Techniques: Immunostaining, Irradiation, Western Blot, Expressing, Co-Culture Assay

Journal: Experimental & Molecular Medicine

Article Title: Megakaryocytic TGFβ1 orchestrates osteogenesis of LepR + SSCs to alleviate radiation-induced bone loss

doi: 10.1038/s12276-025-01612-z

Figure Lengend Snippet: a Representative micro-CT images of longitudinal section femurs, cross-sectional view of the distal femurs and reconstructed trabecular structure of the region of interest from Slc39a14 leprΔ/Δ mice and their littermate controls (Slc39a14 fl/fl mice) ( n = 6 mice per group). b Quantitative micro-CT analysis of the TB fraction (BV/TV, Tb.N, Tb.Th, Tb.Sp, BMD and Ct.Th) in Slc39a14 leprΔ/Δ mice and their littermate controls (Slc39a14 fl/fl mice) ( n = 6 mice per group). c Quantitative biomechanical analysis of femora (peak load and stiffness) from MK deleted mice and their littermate controls (Slc39a14 fl/fl mice) ( n = 6 mice per group). d Serum zinc concentration in Slc39a14 leprΔ/Δ mice and their littermate controls (Slc39a14 fl/fl mice) ( n = 6 per group). e Von Kossa staining showing mineralization of bone matrix in Slc39a14 leprΔ/Δ mice and their littermate controls (Slc39a14 fl/fl mice) ( n = 6 per group). f Representative micro-CT images of longitudinal section femurs, cross-sectional view of the distal femurs and reconstructed trabecular structure of the region of interest from Slc39a14 leprΔ/Δ mice injected with TPO or vehicle after irradiation ( n = 6 mice per group). g Representative immunostaining images of OCN (red) in the TB and EB of the Slc39a14 leprΔ/Δ mice injected with TPO or vehicle after irradiation. The quantification of OCN cells is shown on the right ( n = 6 mice per group). Scale bar, 100 µm. h Representative immunostaining images of LepR (red) and PTP1B (green) in the BM of the Slc39a14 leprΔ/Δ mice injected with TPO or vehicle after irradiation. Scale bar, 100 µm. i Colocalization of LepR (red) with Ki67 (green) in the BM of the Slc39a14 leprΔ/Δ mice injected with TPO or vehicle after irradiation ( n = 6 mice per group). Scale bar, 100 µm. j Representative immunostaining images of TUNEL (green) in the BM of the Slc39a14 leprΔ/Δ mice injected with TPO or vehicle after irradiation. The quantification of tunel positive cells is shown on the right ( n = 6 mice per group). Scale bar, 100 µm. Data on graphs are shown as mean ± SD. An unpaired two-tailed t -test was used to analyze the data in b – d , g , i and j . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.

Article Snippet: Briefly, the bone sections were incubated with primary antibodies against mouse osteocalcin (A20800, ABclonal), Ki67 (AF7617, R&D), PTP1B (bs-55182R, Bioss), Slc39a14 (A10413, ABclonal), leptin receptor (bs-0410R, Bioss), CHOP ( A21902 , ABclonal), F4/80 (30325, CST), TGFβ1 (ab313729, abcam), vWF (bsm-52775R, Bioss), osterix (ab209484, abcam) and Smad2 (A7699, ABclonal) overnight at 4 °C and incubated with secondary antibodies for 1 h at 37 °C.

Techniques: Micro-CT, Concentration Assay, Staining, Injection, Irradiation, Immunostaining, TUNEL Assay, Two Tailed Test

Journal: Translational Oncology

Article Title: The CHPT-pSTAT3-SLC7A11 signaling axis controls progression and ferroptosis susceptibility of pancreatic cancer

doi: 10.1016/j.tranon.2025.102624

Figure Lengend Snippet: CHPT1 may regulate SLC7A11 transcription by phosphorylating STAT3. A. The results of protein profiling after immunoprecipitation performed by CHPT1 were intersected with the list of transcription factors predicted for SLC7A11 taken to find STAT3. B. Immunoprecipitation (Co-IP) assay to verify the direct interaction of CHPT1 with STAT3. C-D Representative images of immunofluorescence staining showing the colocalization of CHPT1 (red) and STAT3 (green) in PDAC cells. DAPI: blue, nucleus. Line chart of fluorescence signal positioning analysis. E. Protein expression levels of phosphorylated Y705 STAT3 after knockdown and overexpression of CHPT1. F. Protein expression levels of SLC7A11 after treatment with Stattic, an inhibitor of phosphorylated STAT3-Y705 to knockdown CHPT1 cells(5μM and 10μM). G. Co-IP assay confirmed the physical binding of PTPN1 to STAT3. H. CHPT1 overexpression promotes the binding of PTPN1 to STAT3. IP was performed under WB quantification and the differences were significant. I. Immunocytofluorescence analysis showed that overexpression of CHPT1 inhibited the nuclear translocation of pSTAT3.

Article Snippet: After clearing non-specific binding proteins with protein A/G-agarose beads (Selleck, USA), cell lysates were incubated with primary antibodies against CHPT1 (1:100, Santa), STAT3 (1:100, Proteintech), PTPN1(1:100, Proteintech), and IgG control (1:100, Proteintech) overnight at 4°C.

Techniques: Immunoprecipitation, Co-Immunoprecipitation Assay, Immunofluorescence, Staining, Fluorescence, Expressing, Knockdown, Over Expression, Binding Assay, Translocation Assay

Journal: Translational Oncology

Article Title: The CHPT-pSTAT3-SLC7A11 signaling axis controls progression and ferroptosis susceptibility of pancreatic cancer

doi: 10.1016/j.tranon.2025.102624

Figure Lengend Snippet: PTPN1 is the critical effector of CHPT1 in suppressing STAT3-driven PDAC progression. A. Western blot analysis confirming efficient knockdown of PTPN1 in CHPT1-overexpressing PDAC cells. B-D. EDU incorporation assays showing that PTPN1 depletion restores cell proliferation despite CHPT1 overexpression. C. CCK-8 assays demonstrating that PTPN1 knockdown rescues the proliferative capacity of CHPT1-overexpressing cells. E. Transwell migration assays revealing the restoration of migratory potential upon PTPN1 silencing in CHPT1-overexpressing cells. F. Wound healing assays further confirming that PTPN1 depletion reverses the anti-migratory effects of CHPT1 overexpression. All data are presented as mean ± SD from 3 independent experiments (n=3).

Article Snippet: After clearing non-specific binding proteins with protein A/G-agarose beads (Selleck, USA), cell lysates were incubated with primary antibodies against CHPT1 (1:100, Santa), STAT3 (1:100, Proteintech), PTPN1(1:100, Proteintech), and IgG control (1:100, Proteintech) overnight at 4°C.

Techniques: Western Blot, Knockdown, Over Expression, CCK-8 Assay, Migration

Journal: Journal of Ginseng Research

Article Title: (20S)-protopanaxatriol attenuates Ang II-induced renal injury via PTPN1-mediated AMPK/mTOR signaling pathway

doi: 10.1016/j.jgr.2025.08.009

Figure Lengend Snippet: The potential target proteins of PPT were determined through network analysis and the effect of PPT on autophagy of renal cells was confirmed (A) In silico prediction of PPT's molecular targets was performed using web-based tools. (B) Molecular docking of PPT with PTPN1. (C) PPT treatment increases protease susceptibility of the PTPN1 in cell lysates as determined by the DARTS assay. (D) NRK-52E cells were treated with 6 μM PPT for 2 h and subsequently heated at different temperatures for 3 min. After freeze-thaw cycles for cell lysis, the soluble PTPN1 protein levels were examined by Western blotting. (E) Western blotting was used to detect the expression of PTPN1 in the renal tissue. GAPDH was included as a loading control. (F) Densitometric quantification of electrophoretic bands from panel E using ImageJ analytical modules. Datasets expressed as mean ± SEM (n = 6 biological replicates). Statistical comparisons performed using two-tailed t -test: ns = non-significant; ∗ p < 0.05, p < 0.01 vs control cohort; # p < 0.05, ## p < 0.01 vs Ang II-treated group.

Article Snippet: Antibodies against PTPN1 (Cat. #11334-1-AP), P62/SQSTM1 (Cat. # 18420-1-AP), Beclin 1 (Cat. # 11306-1-AP), LC3 (Cat. # 14600-1-AP), IκBα (Cat. # 10268-1-AP), p-P65 Ser468 (Cat. # 14600-1-AP) were procured from Proteintech (China). p-AMPKα Thr172 (Cat. #2535), AMPKα (Cat. #2532), p-mTOR Ser2448 (Cat. #5536) and mTOR (Cat. #2983) were procured from Cell Signaling Technology (USA).

Techniques: In Silico, Lysis, Western Blot, Expressing, Control, Two Tailed Test

Journal: Journal of Ginseng Research

Article Title: (20S)-protopanaxatriol attenuates Ang II-induced renal injury via PTPN1-mediated AMPK/mTOR signaling pathway

doi: 10.1016/j.jgr.2025.08.009

Figure Lengend Snippet: PPT reduces Ang II‐induced fibrosis in NRK-52E cells NRK-52E renal tubular epithelial cells underwent a pretreatment protocol with 5 or 10 μM PPT for 1 h prior to Ang II stimulation (1 μM, 8/24 h). (A) Cytocompatibility assessment via colorimetric MTT cell viability assay (n = 3 technical replicates). (B) Immunoblot detection of fibrotic biomarkers COL-IV and TGF-β1 with GAPDH normalization. (C) Densitometric quantification of electrophoretic bands from panel B using ImageJ analytical modules. (D) Quantitative gene expression analysis of Col4 and Tgfb transcripts via RT-qPCR amplification. (E) Western blotting was used to detect the expression of PTPN1 in the NRK-52E cells. GAPDH was included as a loading control. (F) Densitometric quantification of electrophoretic bands from panel E using ImageJ analytical modules. (G – I) Inflammatory cytokine transcript quantification ( Il1b, Il6, Tnf ) in NRK-52E cells by reverse transcription-quantitative PCR. (J) Pharmacological interrogation protocol: NRK-52E cultures received 1 h pretreatment with PPT (5/10 μM) ± PTPN1 inhibitor cocktail (5 μM: Trodusquemine) followed by 2 h Ang II stimulation (1 μM). Subsequent biomolecular extraction facilitated parallel immunoblotting and transcriptional analysis. Representative immunodetection of fibrotic effectors COL-IV and TGF-β1 with GAPDH normalization. (K) Densitometric quantification of panel J immunoblots. (L – N) Inflammatory transcriptome mapping of Il1b, Il6, and Tnf via quantitative reverse transcription PCR. All data expressed as mean ± SEM (n = 3 biological experiments). Statistical significance determined by two-tailed t -test: ∗ p < 0.05, ∗∗ p < 0.01 vs untreated controls; # p < 0.05, ## p < 0.01 vs Ang II-challenged cells.

Article Snippet: Antibodies against PTPN1 (Cat. #11334-1-AP), P62/SQSTM1 (Cat. # 18420-1-AP), Beclin 1 (Cat. # 11306-1-AP), LC3 (Cat. # 14600-1-AP), IκBα (Cat. # 10268-1-AP), p-P65 Ser468 (Cat. # 14600-1-AP) were procured from Proteintech (China). p-AMPKα Thr172 (Cat. #2535), AMPKα (Cat. #2532), p-mTOR Ser2448 (Cat. #5536) and mTOR (Cat. #2983) were procured from Cell Signaling Technology (USA).

Techniques: Viability Assay, Western Blot, Gene Expression, Quantitative RT-PCR, Amplification, Expressing, Control, Reverse Transcription, Real-time Polymerase Chain Reaction, Extraction, Immunodetection, Two Tailed Test

Journal: Journal of Ginseng Research

Article Title: (20S)-protopanaxatriol attenuates Ang II-induced renal injury via PTPN1-mediated AMPK/mTOR signaling pathway

doi: 10.1016/j.jgr.2025.08.009

Figure Lengend Snippet: Graphical abstract mechanism of (20S)-Protopanaxatriol in alleviating hypertensive inflammatory kidney injury by targeting the PTPN1-Mediated AMPK/mTOR autophagy signaling pathway.

Article Snippet: Antibodies against PTPN1 (Cat. #11334-1-AP), P62/SQSTM1 (Cat. # 18420-1-AP), Beclin 1 (Cat. # 11306-1-AP), LC3 (Cat. # 14600-1-AP), IκBα (Cat. # 10268-1-AP), p-P65 Ser468 (Cat. # 14600-1-AP) were procured from Proteintech (China). p-AMPKα Thr172 (Cat. #2535), AMPKα (Cat. #2532), p-mTOR Ser2448 (Cat. #5536) and mTOR (Cat. #2983) were procured from Cell Signaling Technology (USA).

Techniques: