p38 mapk polyclonal antibody  (Proteintech)

Journal: Journal of Traditional and Complementary Medicine

Article Title: Osthole attenuates trastuzumab-induced cardiotoxicity in mice by enhancing autophagy via regulating the p38MAPK/mTOR signaling pathway

doi: 10.1016/j.jtcme.2025.06.001

Figure Lengend Snippet: Effects of OST on cardiac autophagy and the p38MAPK/mTOR signaling pathway in TRZ-treated mice. (A–B) Representative images of transmission electron microscopy images, yellow arrow, mitochondria; red arrow, autophagosome. (C) Representative western blotting bands for LC3I, LC3II, Beclin-1, p62. (D–F) Quantitative analysis of LC3II/I, Beclin-1, p62 (normalized to GAPDH). (G) Representative western blotting bands for p-p38MAPK, p38MAPK, p-mTOR and mTOR. (H) Quantitative analysis of p-p38MAPK/p38MAPK and p-mTOR/mTOR. Data are expressed as means ± SEM ( n = 3). ## P < 0.01 vs the CON group; ∗ P < 0.05, ∗∗ P < 0.01 vs the TRZ group.

Article Snippet: The first antibodies to Bax (1:1000 dilution, A0207), Bcl-2 (1:800 dilution, A0208), Caspase-3 (1:800 dilution, A2156), LC3 (1:500 dilution, A19665), Beclin-1 (1:1000 dilution, A7353), p62 (1:1000 dilution, A19700) and GAPDH (1:800 dilution, A19056) were purchased from ABclonal Company (Wuhan, China), p38MAPK (1:1000 dilution, bs-0637R) and p-p38MAPK (1:1000 dilution, bs-0636R) were purchased from Bioss Company (Beijing, China), mTOR (1:1000 dilution, #2983) and p-mTOR (1:1000 dilution, #2971) were purchased from CST Company (Danvers, MA, USA).

Techniques: Transmission Assay, Electron Microscopy, Western Blot

Journal: Journal of Traditional and Complementary Medicine

Article Title: Osthole attenuates trastuzumab-induced cardiotoxicity in mice by enhancing autophagy via regulating the p38MAPK/mTOR signaling pathway

doi: 10.1016/j.jtcme.2025.06.001

Figure Lengend Snippet: Effects of OST on autophagy and the p38MAPK/mTOR signaling pathway in TRZ-treated mice after SB203580 administration. (A–B) Representative images of transmission electron microscopy images, yellow arrow, mitochondria; red arrow, autophagosome. (C) Representative western blotting bands for LC3I, LC3II, Beclin-1, p62. (D–F) Quantitative analysis of LC3II/I, Beclin-1, p62 (normalized to GAPDH). (G) Representative western blotting bands for p-mTOR and mTOR. (H) Quantitative analysis of p-mTOR/mTOR. Data are expressed as means ± SEM ( n = 3). # P < 0.05, ## P < 0.01 vs the CON group; ∗ P < 0.05, ∗∗ P < 0.01 vs the TRZ group; ▲ P < 0.05, ▲▲ P < 0.01 vs the TRZ + OST H group.

Article Snippet: The first antibodies to Bax (1:1000 dilution, A0207), Bcl-2 (1:800 dilution, A0208), Caspase-3 (1:800 dilution, A2156), LC3 (1:500 dilution, A19665), Beclin-1 (1:1000 dilution, A7353), p62 (1:1000 dilution, A19700) and GAPDH (1:800 dilution, A19056) were purchased from ABclonal Company (Wuhan, China), p38MAPK (1:1000 dilution, bs-0637R) and p-p38MAPK (1:1000 dilution, bs-0636R) were purchased from Bioss Company (Beijing, China), mTOR (1:1000 dilution, #2983) and p-mTOR (1:1000 dilution, #2971) were purchased from CST Company (Danvers, MA, USA).

Techniques: Transmission Assay, Electron Microscopy, Western Blot

Journal: Journal of Traditional and Complementary Medicine

Article Title: Osthole attenuates trastuzumab-induced cardiotoxicity in mice by enhancing autophagy via regulating the p38MAPK/mTOR signaling pathway

doi: 10.1016/j.jtcme.2025.06.001

Figure Lengend Snippet: Schematic illustration summarizing the protective effects of OST against TRZ-induced cardiotoxicity by enhancing autophagy via regulating the p38MAPK/mTOR signaling pathway.

Article Snippet: The first antibodies to Bax (1:1000 dilution, A0207), Bcl-2 (1:800 dilution, A0208), Caspase-3 (1:800 dilution, A2156), LC3 (1:500 dilution, A19665), Beclin-1 (1:1000 dilution, A7353), p62 (1:1000 dilution, A19700) and GAPDH (1:800 dilution, A19056) were purchased from ABclonal Company (Wuhan, China), p38MAPK (1:1000 dilution, bs-0637R) and p-p38MAPK (1:1000 dilution, bs-0636R) were purchased from Bioss Company (Beijing, China), mTOR (1:1000 dilution, #2983) and p-mTOR (1:1000 dilution, #2971) were purchased from CST Company (Danvers, MA, USA).

Techniques:

Journal: Redox Biology

Article Title: Stromal cell-derived itaconate promotes endometriosis via macrophage NRF2 and lysosomal pH modulation

doi: 10.1016/j.redox.2026.104101

Figure Lengend Snippet: Itaconate Regulates Lysosomal Function, Calcium Signaling, and p38 MAPK Pathway in Macrophages (A) Lysosomal acidification in ascites-derived macrophages from non-endometriosis (NC) and endometriosis (EM) patients assessed by LysoSensor fluorescence intensity (n = 6/group). (B) Lysosomal acidification in PBMC-derived macrophages after LPS,IL-4 and 4-OI treatment (n = 3/group). (C) Lysosomal pH value of BMDMs after LPS and 4-OI treatment (n = 3/group). (D, E) Effect of chloroquine (CQ) on LysoSensor intensity (D) and proportion of iNOS + BMDMs (E) after LPS and 4-OI treatment (n = 3/group). (F) mRNA levels of Il1b , Il6 , Nos2 , and Tnf in BMDMs under indicated treatments (n = 3/group). (G) Intracellular Ca 2+ dynamics in BMDMs after treatments, measured by Fluo-4. (H) Intracellular Ca 2+ dynamics in ascites-derived macrophages from NC and EM patients. (I) Quantification of intracellular calcium in peritoneal macrophages from NC and EM patients (n = 3/group). (J) Quantification of intracellular calcium in BMDMs (n = 5/group). (K) mRNA expression of lysosomal calcium channel genes ( MCOLN1 , MCOLN2 , TPC1 , TPC2 ) in ascites-derived macrophages from NC and EM patients (n = 3/group). (L) lysosomal calcium channel genes mRNA in PBMC-derived macrophages co-cultured with normal ESCs (Nor-ESC), eutopic ESCs (Eu-ESC), or ectopic ESCs (Ec-ESC) from patients (n = 5/group). (M − N) Fluo-4-based Ca 2+ dynamics in BMDMs treated with LPS, 4-OI, ML-SA1 (M), or CQ (N). (O) Flow cytometry analysis and quantification of iNOS + BMDMs after indicated treatments (n = 3/group). (P) mRNA levels of pro-inflammatory genes in BMDMs under different treatments (n = 3/group). (Q) Western blot and quantification of p-p38 and total p38 in BMDMs with LPS ± 4-OI (n = 3/group). (R–S) Western blot and quantification of iNOS, p-p38, and total p38 in BMDMs treated with LPS, 4-OI, and CQ (R), or ML-SA1 (S) (n = 3/group). (Data are presented as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.) EM, endometriosis; NC, non-EM control; LPS, lipopolysaccharide; 4-OI, 4-octyl itaconate; CQ, chloroquine; ML-SA1, MCOLN channel agonist; PBMC, peripheral blood mononuclear cell; ESC, endometrial stromal cell.

Article Snippet: After blocking in 5% milk in TBST, membranes were incubated overnight at 4 °C with primary antibodies: Beta Actin Monoclonal antibody(1:20000, 66009-1-Ig; Proteintech), Beta Tubulin Recombinant antibody (1:5000, 80713-1-RR; Proteintech), Anti-IRG1 antibody (1:1000; ab222411; abcam; Cambridge, UK), Nrf2 monoclonal antibody (1:2000; A21176; abclonal; Wuhan, China), iNOS Polyclonal antibody (1:500; 22226-1-AP; Proteintech), NOX2 Polyclonal antibody (1:3000; 19013-1-AP; Proteintech), p-p38 MAPK Polyclonal antibody (1:2000; 28796-1-AP; Proteintech), and p38 MAPK Polyclonal antibody (1:4000; 14064-1-AP; Proteintech).

Techniques: Derivative Assay, Fluorescence, Expressing, Cell Culture, Flow Cytometry, Western Blot, Control

Journal: Redox Biology

Article Title: Stromal cell-derived itaconate promotes endometriosis via macrophage NRF2 and lysosomal pH modulation

doi: 10.1016/j.redox.2026.104101

Figure Lengend Snippet: Itaconate Regulates Lysosomal Function, Calcium Signaling, and p38 MAPK Pathway in Macrophages (A) Lysosomal acidification in ascites-derived macrophages from non-endometriosis (NC) and endometriosis (EM) patients assessed by LysoSensor fluorescence intensity (n = 6/group). (B) Lysosomal acidification in PBMC-derived macrophages after LPS,IL-4 and 4-OI treatment (n = 3/group). (C) Lysosomal pH value of BMDMs after LPS and 4-OI treatment (n = 3/group). (D, E) Effect of chloroquine (CQ) on LysoSensor intensity (D) and proportion of iNOS + BMDMs (E) after LPS and 4-OI treatment (n = 3/group). (F) mRNA levels of Il1b , Il6 , Nos2 , and Tnf in BMDMs under indicated treatments (n = 3/group). (G) Intracellular Ca 2+ dynamics in BMDMs after treatments, measured by Fluo-4. (H) Intracellular Ca 2+ dynamics in ascites-derived macrophages from NC and EM patients. (I) Quantification of intracellular calcium in peritoneal macrophages from NC and EM patients (n = 3/group). (J) Quantification of intracellular calcium in BMDMs (n = 5/group). (K) mRNA expression of lysosomal calcium channel genes ( MCOLN1 , MCOLN2 , TPC1 , TPC2 ) in ascites-derived macrophages from NC and EM patients (n = 3/group). (L) lysosomal calcium channel genes mRNA in PBMC-derived macrophages co-cultured with normal ESCs (Nor-ESC), eutopic ESCs (Eu-ESC), or ectopic ESCs (Ec-ESC) from patients (n = 5/group). (M − N) Fluo-4-based Ca 2+ dynamics in BMDMs treated with LPS, 4-OI, ML-SA1 (M), or CQ (N). (O) Flow cytometry analysis and quantification of iNOS + BMDMs after indicated treatments (n = 3/group). (P) mRNA levels of pro-inflammatory genes in BMDMs under different treatments (n = 3/group). (Q) Western blot and quantification of p-p38 and total p38 in BMDMs with LPS ± 4-OI (n = 3/group). (R–S) Western blot and quantification of iNOS, p-p38, and total p38 in BMDMs treated with LPS, 4-OI, and CQ (R), or ML-SA1 (S) (n = 3/group). (Data are presented as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.) EM, endometriosis; NC, non-EM control; LPS, lipopolysaccharide; 4-OI, 4-octyl itaconate; CQ, chloroquine; ML-SA1, MCOLN channel agonist; PBMC, peripheral blood mononuclear cell; ESC, endometrial stromal cell.

Article Snippet: After blocking in 5% milk in TBST, membranes were incubated overnight at 4 °C with primary antibodies: Beta Actin Monoclonal antibody(1:20000, 66009-1-Ig; Proteintech), Beta Tubulin Recombinant antibody (1:5000, 80713-1-RR; Proteintech), Anti-IRG1 antibody (1:1000; ab222411; abcam; Cambridge, UK), Nrf2 monoclonal antibody (1:2000; A21176; abclonal; Wuhan, China), iNOS Polyclonal antibody (1:500; 22226-1-AP; Proteintech), NOX2 Polyclonal antibody (1:3000; 19013-1-AP; Proteintech), p-p38 MAPK Polyclonal antibody (1:2000; 28796-1-AP; Proteintech), and p38 MAPK Polyclonal antibody (1:4000; 14064-1-AP; Proteintech).

Techniques: Derivative Assay, Fluorescence, Expressing, Cell Culture, Flow Cytometry, Western Blot, Control

Journal: International Dental Journal

Article Title: 4632427E13Rik Facilitates Jaw Marrow-Derived Mesenchymal Stem Cells Osteogenesis and Angiogenesis Under Hypoxia Through miR-34a-5p/Aldoa/Hif-1α Pathway

doi: 10.1016/j.identj.2025.109364

Figure Lengend Snippet: Aldoa promotes osteogenesis and angiogenesis via the ERK/Hif-1α pathway. A, Alizarin red staining (14 days) and ALP staining (7 days). B, The gene expression levels of Runx2, Alp and Ocn were determined by qRT-PCR. β-actin was used as an internal reference gene. C, Tube formation assay was performed in the presence of CM. The scale bars represent 100 μm. D, The mRNA expression levels of Vegf and CD31 were analysed by quantitative RT-PCR. β-actin was used as an internal reference gene. E, Immunofluorescence staining was performed to detect Vegf expression after being treated by CM. The scale bars represent 25 μm. F, A Kyoto Encyclopaedia of Genes and Genomes (KEGG) enrichment analysis was performed to determine the top related pathways involving these differentially expressed mRNAs. G and H, The protein expression levels of Aldoa, Hif-1α and the phosphorylated of ERK, JUK and p38 among hypoxia, normoxia and hypoxia + si-Aldoa groups were determined by western blot. Semiquantitative analysis of the p-ERK/ERK, p-JUK/JUK and p38/p-p38 ratios were shown. β-actin was used as an internal reference gene. All data were expressed as means ± SD. * P < .05, ** P < .01.

Article Snippet: Rabbit anti-Aldoa (1:1000, 11217-1-AP), rabbit anti-Hif-1α (1:1000, ab179483), rabbit anti-Runx2 (1:1000, 8486S), rabbit anti-CD31 (1:1000, 77699S), rabbit anti-Ocn (1:1000, bs-4917R), rabbit anti-Alp (1:1000, bs-1535R), rabbit anti-Vegf (1:1000, bs-1313R), rabbit anti-ERK (1:1000, bsm-33337M), rabbit anti-p-ERK (1:1000, bs-1646R), rabbit anti-p38 (1:1000, bs-0637R), rabbit anti-p-p38 (1:1000, bs-0636R), rabbit anti-JNK (1:1000, bs-2592R), rabbit anti-p-JNK (1:1000, bs-1640R), and mouse anti-β-actin (1:2000, AF7018) were purchased from Proteintech, Abcam, Cell Signalling Technology, and Bioss, respectively.

Techniques: Staining, Gene Expression, Quantitative RT-PCR, Tube Formation Assay, Expressing, Immunofluorescence, Western Blot