Journal: PLoS ONE

Article Title: CD45RB Is a Novel Molecular Therapeutic Target to Inhibit Aβ Peptide-Induced Microglial MAPK Activation

doi: 10.1371/journal.pone.0002135

Figure Lengend Snippet: Microglial treatment conditions are indicated and are further described in . Cell lysates were analyzed by Western immunoblotting using specific antibodies that recognize phosphorylated or total p38 MAPK and/or p44/42 MAPK at the indicated time points (A and B, top panel). Phosphorylation of both p38 MAPK and/or p44/42 MAPK after treatment with LPS was inhibited by SB203580 or PD98059 (A and B, bottom panel). Histograms below the immunoblots represent the mean band density ratio±1 SD (pp38 MAPK/total p38 MAPK and/or pp44/42 MAPK/total p44/42 MAPK; n = 3 for each condition presented; * P <0.05, ** P <0.001 compared with control). (C) Microglial phagocytosis of Aβ 1–42 peptide after pre-treatment with PD98059 or SB203580 for 1 h, then co-treated with “aged” FITC-tagged Aβ 1–42 and LPS. Supernatants and cell lysates were analyzed for extracellular (top panel) and cell-associated (bottom panel) FITC-Aβ 1–42 using a fluorometer (* P <0.05, ** P <0.001). (D) Phosphorylation of p38 MAPK and inhibition of this effect by CD45RB antibody. Histograms below the immunoblots represent the mean band density ratio±1 SD (pp38/total p38; n = 3 for each condition presented; * P <0.05 compared with LPS or LPS/IgG). (E) Microglial activation is evidenced by mean TNF-α and IL-6 release±1 SD (n = 3 for each condition presented; * P <0.05; ** P <0.001 compared with LPS or LPS/IgG; P <0.05 compared LPS/SB with LPS/CD45RB Aβ/SB). For A–E, one-way ANOVA followed by post hoc Bonferroni testing was utilized. Note: SB = SB203580, PD = PD98059, Ab = antibody, pp = phosphorylatioin.

Article Snippet: Antibodies for phospho-p44/42 (pp44/42, Thr202/Tyr204) MAPK, phospho-p38 (pp38, Thr180/Tyr182) MAPK, total p44/42 MAPK, and total p38 MAPK were obtained from Cell Signaling Technology (Beverly, MA) as well as cell lysis buffer, and SDS blue loading buffers.

Techniques: Western Blot, Inhibition, Activation Assay

Journal: PLoS ONE

Article Title: CD45RB Is a Novel Molecular Therapeutic Target to Inhibit Aβ Peptide-Induced Microglial MAPK Activation

doi: 10.1371/journal.pone.0002135

Figure Lengend Snippet: (A) Microglial cells were treated as indicated for 16 h. Microglial activation was determined by TNF-α and IL-6 production (mean±1 SD, picograms per milligram of total protein) in cultured media (top and bottom panels). Data are representative of three independent experiments (** P <0.001). (B–D) Cell lysates were analyzed by Western immunoblotting using specific antibodies that recognize phosphorylated p38 MAPK at the indicated time points. Phosphorylation of p38 MAPK is indicated in (B) and inhibition of this effect by CD45RB Ab is indicated in Fig C or by SB in Fig. D. Histograms below the immunoblots represent the mean band density ratio±1 SD (p38/total p38 MAPK; n = 3 for each condition presented). (E) Microglial cells were pre-treated with SB230580 or CD45RB antibody for 1 h, then co-treated with phen and/or aged Aβ 1–42 for 16 h. Microglial activation was determined by TNF-α and IL-6 production (mean±1 SD) in cultured media (top and bottom panels; n = 3 for each condition presented). One-way ANOVA followed by post hoc Bonferroni testing revealed significant differences (* P <0.05, ** P <0.001). Note: SB = SB203580, Ab = antibody, pp = phosphorylation).

Article Snippet: Antibodies for phospho-p44/42 (pp44/42, Thr202/Tyr204) MAPK, phospho-p38 (pp38, Thr180/Tyr182) MAPK, total p44/42 MAPK, and total p38 MAPK were obtained from Cell Signaling Technology (Beverly, MA) as well as cell lysis buffer, and SDS blue loading buffers.

Techniques: Activation Assay, Cell Culture, Western Blot, Inhibition

Journal: Canadian journal of physiology and pharmacology

Article Title: Effects of somatostatin, curcumin, and quercetin on the fatty acid profile of breast cancer cell membranes.

doi: 10.1139/cjpp-2019-0352

Figure Lengend Snippet: Figure 3: Absorbances of the MAPK pathway members; AKT1, p-AKT-1(Ser473), p-AKT-1 (Thr308), p- P44/42 MAPK (Thr202/Tyr204), S6-Ribozomal and p-S6-Ribozomal (Ser235/236) measured in the MCF-7 cells after incubations Somatostatin, Curcumin, Somatostatin+Curcumin, Quercetin or Somatostatin+Quercetin for 24 h (n=6.*,**Values significantly different from the control p <0.05, p < 0.01. &,&& Values significantly different from the Quercetin p<0.05, p <0.01. #,## Values significantly different from the Somatostatin p <0.05, p <0.01. +,+ +Values significantly different from the Curcumin p <0.05, p <0.01

Article Snippet: Cells were lysed with Cell Lysis Buffer (Cell Signaling Technology), then the levels of AKT1, pAKT-1(Ser473), p-AKT-1 (Thr308), p-P44/42 MAPK (Thr202/Tyr204), S6-Ribozomal and p-S6Ribozomal (Ser235/236) in the cell lysates were determined using the PathScan MAPK Multi-Target Sandwich ELISA Kit (Cell Signaling Technology) according to the manufacturer's instructions.

Techniques: Control

Journal: Canadian journal of physiology and pharmacology

Article Title: Effects of somatostatin, curcumin, and quercetin on the fatty acid profile of breast cancer cell membranes.

doi: 10.1139/cjpp-2019-0352

Figure Lengend Snippet: Figure 4: Absorbances of the MAPK pathway members; AKT1, p-AKT-1(Ser473), p-AKT-1 (Thr308), p- P44/42 MAPK (Thr202/Tyr204), S6-Ribozomal and p-S6-Ribozomal (Ser235/236) measured in the MDA- MB231 cells after incubations Somatostatin, Curcumin, Somatostatin+Curcumin, Quercetin or Somatostatin+Quercetin for 24 h (n=6). .*,**Values significantly different from the control p <0.05, p < 0.01. &,&& Values significantly different from the Quercetin p<0.05, p <0.01. #,## Values significantly different from the Somatostatin p <0.05, p <0.01. +,+ +Values significantly different from the Curcumin p <0.05, p <0.01

Article Snippet: Cells were lysed with Cell Lysis Buffer (Cell Signaling Technology), then the levels of AKT1, pAKT-1(Ser473), p-AKT-1 (Thr308), p-P44/42 MAPK (Thr202/Tyr204), S6-Ribozomal and p-S6Ribozomal (Ser235/236) in the cell lysates were determined using the PathScan MAPK Multi-Target Sandwich ELISA Kit (Cell Signaling Technology) according to the manufacturer's instructions.

Techniques: Control