Journal: PLoS ONE

Article Title: Increasing Hippocampal Estrogen Receptor Alpha Levels via Viral Vectors Increases MAP Kinase Activation and Enhances Memory in Aging Rats in the Absence of Ovarian Estrogens

doi: 10.1371/journal.pone.0051385

Figure Lengend Snippet: Western blot data showing the effects of Lenti-ERα or Lenti-Cherry on levels of phosphorylated p44 MAPK and phosphorylated p42 MAPK (A), and phosphorylated Akt (B) in the hippocampus. Mean density x area (+SEM) expressed relative to Lenti-Cherry control. * P = .043 vs. Lenti-Cherry. Representative blot images for phosphorylated and total p44 MAPK, p42 MAPK, and Akt are shown in insets above the graph.

Article Snippet: Blots were then blocked and incubated with primary antibodies for p42/p44 MAPK (rabbit polyclonal, 1∶4000; Cell Signaling), Akt (rabbit polyclonal, 1∶1000; Cell Signaling), or β-actin (mouse monoclonal, 1∶15,000; Santa Cruz) overnight at 4°C in 1% nonfat dry milk-TTBS.

Techniques: 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: Stem Cells International

Article Title: Interleukin-1 β Enhances Umbilical Cord Mesenchymal Stem Cell Adhesion Ability on Human Umbilical Vein Endothelial Cells via LFA-1/ICAM-1 Interaction

doi: 10.1155/2019/7267142

Figure Lengend Snippet: p38 MAPK signaling pathway is involved in IL-1 β -mediated LFA-1 expression in MSCs and MSC adhesion to HUVECs. (a) Immunocytochemistry staining for LFA-1 (green) and DAPI (blue) in MSCs. Cells were treated with p38 MAPK (SB 203580, 5 μ M), AKT (GSK690693, 20 μ M), ERK1/2 (U0126 20 μ M), and JNK (SP600125 20 nM) inhibitor and combined with IL-1 β for 30 minutes (scale bar: 50 μ m). (b) Western blot results of the p38 MAPK and phosphorylated p38 MAPK from the lysates of cells pretreated with IL-1 β at 0, 1, 3, 5, 10, and 20 minutes. (c) Western blot results of the LFA-1 (129 kDa) expression in membrane fractions of MSCs treated with IL-1 β and SB 203580 (SB) or cotreated with both IL-1 β and SB. (d) Quantitative graphs of the Western blot results of LFA-1 expression of (c) ( n = 3, ∗∗ P < 0.01). (e) Representative image of MSC adhesion on HUVECs. MSCs were treated with IL-1 β and inhibitor SB 203580 adhesion to IL-1 β -activated HUVECs or nonactivated HUVECs. MSCs labelled with calcein AM (5 μ M) (green), HUVECs, and MSC cell nuclei were stained with Hoechst 33258 (blue) (scale bar: 50 μ m). (f) Quantitative graphs of the cell adhesion assay results of MSCs treated with IL-1 β and MAPK inhibitor SB 203580 adhesion to nonactivated HUVECs (black bars) or IL-1 β -activated HUVECs (gray bars). Values were the cell number fold change relative to the control group. Data represent mean ± SD ( n = 3, ∗∗∗ P < 0.005, ∗∗ P < 0.01, and ∗ P < 0.05) (N.S.: nonsignificance).

Article Snippet: The membrane was blocked in 5% fish gelatin blocking buffer (Amresco, OH, USA) for 1 hour and then incubated with the anti-human CD11a antibody (GeneTex, USA) at 1 : 4000 dilution, anti-p38 MAPK antibody (Santa Cruz, TX, USA) at 1 : 200 dilution, and phospho-p38 MAPK primary antibodies (Cell Signaling, MA, USA) at 4°C overnight.

Techniques: Expressing, Immunocytochemistry, Staining, Western Blot, Membrane, Cell Adhesion Assay, Control

Journal: Stem Cells International

Article Title: Interleukin-1 β Enhances Umbilical Cord Mesenchymal Stem Cell Adhesion Ability on Human Umbilical Vein Endothelial Cells via LFA-1/ICAM-1 Interaction

doi: 10.1155/2019/7267142

Figure Lengend Snippet: Schematic diagram of IL-1 β signaling pathway in MSC adhesion to HUVECs. The schematic diagram depicts the proposed role of IL-1 β signaling pathway in MSC adhesion to HUVECs. The process of cell adhesion is initiated by IL-1 β through p38 MAPK; induced expression of LFA-1 in MSCs enhances the cell adhesion to IL-1 β -induced ICAM-1 in HUVECs.

Article Snippet: The membrane was blocked in 5% fish gelatin blocking buffer (Amresco, OH, USA) for 1 hour and then incubated with the anti-human CD11a antibody (GeneTex, USA) at 1 : 4000 dilution, anti-p38 MAPK antibody (Santa Cruz, TX, USA) at 1 : 200 dilution, and phospho-p38 MAPK primary antibodies (Cell Signaling, MA, USA) at 4°C overnight.

Techniques: Expressing