Journal: PLoS ONE

Article Title: Sexual Dimorphism in MAPK-Activated Protein Kinase-2 (MK2) Regulation of RANKL-Induced Osteoclastogenesis in Osteoclast Progenitor Subpopulations

doi: 10.1371/journal.pone.0125387

Figure Lengend Snippet: Cells were fixed, TRAP stained, and visualized by light microscopy. (A) Representative 100x images of female (top) and male (bottom) mouse bone marrow cells defined by CD11b surface expression driven to form osteoclasts (OCs) with M-CSF (25ng/mL) and RANKL (50 ng/mL). Female OCs (black arrow) were magnified 3x and inset (top). TRAP positive cells with three or more nuclei (OCs) from (B) female and (C) male mice were enumerated from three random images at days 3 (top) and 5 (bottom) of RANKL treatment. Osteoclast size from female (B, right) and male (C, right) was measured by pixels per osteoclast using Adobe PhotoShop CS. Data are expressed as means ± SE compared to Mk2 +/+ controls (* P ≤0.05, ** P ≤0.01, *** P ≤0.001).

Article Snippet: Membranes were incubated in Cell Signaling primary antibodies: p-MK2 (Thr334, 27B7, rabbit mAb), p-p38 (Thr180/Tyr182, rabbit polyclonal), p38 (rabbit polyclonal), and GAPDH (14C10, rabbit mAb) at a 1:1000 dilution in 5% BSA in TBS-T overnight at 4°C.

Techniques: Staining, Light Microscopy, Expressing

Journal: PLoS ONE

Article Title: Sexual Dimorphism in MAPK-Activated Protein Kinase-2 (MK2) Regulation of RANKL-Induced Osteoclastogenesis in Osteoclast Progenitor Subpopulations

doi: 10.1371/journal.pone.0125387

Figure Lengend Snippet: (A) Representative flow cytometry of bone marrow from male and female Mk2 +/+ and Mk2 -/- mice. (B) Enumeration of Gr-1 - CD11b lo cells as a percent of the hematopoietic stem cell population (HSC, top) and CD115+ as a percent of Gr-1 - CD11b lo cells (bottom). (C) Enumeration of CD11b surface expression in female (left) and male (right) mouse bone marrow cells. Data are expressed as means ± SE compared to Mk2 +/+ controls (* P ≤0.05).

Article Snippet: Membranes were incubated in Cell Signaling primary antibodies: p-MK2 (Thr334, 27B7, rabbit mAb), p-p38 (Thr180/Tyr182, rabbit polyclonal), p38 (rabbit polyclonal), and GAPDH (14C10, rabbit mAb) at a 1:1000 dilution in 5% BSA in TBS-T overnight at 4°C.

Techniques: Flow Cytometry, Expressing

Journal: PLoS ONE

Article Title: Sexual Dimorphism in MAPK-Activated Protein Kinase-2 (MK2) Regulation of RANKL-Induced Osteoclastogenesis in Osteoclast Progenitor Subpopulations

doi: 10.1371/journal.pone.0125387

Figure Lengend Snippet: (A) Representative western blot of pre-osteoclasts from male mice primed with M-CSF (10 ng/mL) for two days and stimulated with RANKL (100 ng/mL) for the indicated minutes (min). Densiometric analysis of (B) p38 (left) and (C) p-p38 (right) as a percent of GAPDH. Data are expressed as means ± SE compared to Mk2 +/+ controls (* P ≤0.05).

Article Snippet: Membranes were incubated in Cell Signaling primary antibodies: p-MK2 (Thr334, 27B7, rabbit mAb), p-p38 (Thr180/Tyr182, rabbit polyclonal), p38 (rabbit polyclonal), and GAPDH (14C10, rabbit mAb) at a 1:1000 dilution in 5% BSA in TBS-T overnight at 4°C.

Techniques: Western Blot

Journal: PLoS ONE

Article Title: Sexual Dimorphism in MAPK-Activated Protein Kinase-2 (MK2) Regulation of RANKL-Induced Osteoclastogenesis in Osteoclast Progenitor Subpopulations

doi: 10.1371/journal.pone.0125387

Figure Lengend Snippet: (A) Representative confocal images of male dOCP lo cells treated with M-CSF (control) or M-CSF and RANKL for 30 minutes. A 3x magnified portion of p-p38 was inset (middle,left). (B) Pearson’s r correlation coefficients for NFATc1 and DAPI colocalization. (C) Pearson’s r correlation coefficients for p-p38 and DAPI colocalization. (D) Pearson’s r correlation coefficients for NFATc1 and p-p38 colocalization. Data are expressed as means ± SE compared to Mk2 +/+ controls (* P ≤0.05).

Article Snippet: Membranes were incubated in Cell Signaling primary antibodies: p-MK2 (Thr334, 27B7, rabbit mAb), p-p38 (Thr180/Tyr182, rabbit polyclonal), p38 (rabbit polyclonal), and GAPDH (14C10, rabbit mAb) at a 1:1000 dilution in 5% BSA in TBS-T overnight at 4°C.

Techniques: Control

Journal: Current Issues in Molecular Biology

Article Title: Mitigation of 3.5 GHz Electromagnetic Field-Induced BV2 Microglial Cytotoxicity by Polydeoxyribonucleotide

doi: 10.3390/cimb47060386

Figure Lengend Snippet: Effect of SP600125, SB203580, or PD98059 on 3.5 GHz EMF-induced growth suppression of BV2 cells and phosphorylation of JNK-1/2, p38 MAPK (MK2), or ERK-1/2 in BV2 cells. ( A ) BV2 cells were exposed to 3.5 GHz EMFs without or with SP600125 (25 μM), a JNK-1/2 inhibitor, SB203580 (25 μM), a p38 MAPK inhibitor, or PD98059 (50 μM), an ERK-1/2 inhibitor for 2 h. The number of surviving cells was analyzed using cell count analysis. Data represent the mean ± SE of three independent experiments. * p < 0.01 compared with the values of the control (no 3.5 GHz EMF exposure). # p < 0.01 compared with the values of 3.5 GHz EMF exposure (no drug), as determined by one-way ANOVA followed by Sidak’s post hoc test. ( B ) A representative image of morphological changes in the conditioned cells in ( A ). ( C ) BV2 cells were exposed to 3.5 GHz EMFs without or with SP600125 (25 μM), SB203580 (25 μM), or PD98059 (50 μM) for 2 h. Whole-cell lysates from the conditioned cells were prepared and analyzed using Western blotting with antibodies.

Article Snippet: The primary antibodies, including eukaryotic initiation factor-2α (eIF-2α) (cat. no. 9722), phosphorylated (p)-extracellular signal-regulated protein kinase-1/2 (p-ERK-1/2) (T202/Y204) (cat. no. 9101), ERK-1/2 (cat. no. 9102), p-JNK-1/2 (T183/Y185) (cat. no. 9251), JNK-1/2 (cat. no. 9252), and p-p38 MAPK (T180/Y182) (cat. no. 9211) p38 MAPK (cat. no. 9212), p-MK2 (T334) (cat. no. 3007), and T-MK2 (cat. no. 3042), were obtained from Cell Signaling Technology, Inc. (Beverly, MA, USA).

Techniques: Phospho-proteomics, Cell Counting, Control, Western Blot

Journal: International journal of cancer

Article Title: Inhibition of MK2 suppresses IL-1β, IL-6, and TNF-α dependent colorectal cancer growth

doi: 10.1002/ijc.31191

Figure Lengend Snippet: A) MK2 is constitutively activated in CT26 cells. Serum-starvation (SS) reduces pMK2, but treatment with IL-1β, IL-6, or TNF-α induces MK2 phosphorylation. Intratumoral treatment with IL-1β, IL-6, and TNF-α at levels produced by vehicle tumors resulted in B) representative images of tumors with C) restored tumor size and D) weight. In organ culture supernatants, cytokine treatment increased the MK2i tumor production of E) IL-1β, F) IL-6, and G) TNF-α. Increases were seen in mRNA levels for H) IL-1β, I) IL-6, and J) TNF-α were also seen. N=10 *p < 0.05, **p<0.01, ***p < 0.005.

Article Snippet: Tumor samples were labeled by immunohistochemistry for pMK2 Thr334 (Cell Signaling Technology, Danvers, MA).

Techniques: Produced, Organ Culture

Journal: Aging (Albany NY)

Article Title: Expression of Rab1A is upregulated in human lung cancer and associated with tumor size and T stage

doi: 10.18632/aging.101087

Figure Lengend Snippet: ( A ) IHC staining of human lung cancer tissue and noncancerous tissues. Shown are representative images of stained tumor and non-cancerous tissue sections. Red arrowhead: high Rab1A staining; black arrowhead: low Rab1A staining. Scale bar = 50 μm. ( B ) Box plot graph showing the statistical analysis of Rab1A expression in lung cancer and paired non-cancerous tissues. ( C ) Scatter plot showing Rab1A staining levels in individual tumors as a ratio of Rab1A staining in lung cancer to the paired non-cancerous tissue. ( D ) IHC staining of human lung cancer tissue microarray and paired noncancerous tissues. Shown are stained tumor and non-cancerous tissue sections representative of different histological types. Scale bar = 20 μm. ( E ) Scatter plot showing levels of Rab1A staining in different histological types of lung cancer as a ratio of Rab1A staining in cancer tissues to paired non-cancerous tissue.

Article Snippet: Reagents were obtained from the following sources: antibody for Rab1A, Proteintech Group (Rosemont, IL); antibodies for P-S6K1(T398), S6K1, P-AKT(S473), AKT, P-ERK(T202/Y204), ERK, P-MK2(T334), MK2 and Tubulin, Cell Signaling Technology (Danvers, MA).

Techniques: Immunohistochemistry, Staining, Expressing, Microarray

Journal: Aging (Albany NY)

Article Title: Expression of Rab1A is upregulated in human lung cancer and associated with tumor size and T stage

doi: 10.18632/aging.101087

Figure Lengend Snippet: Rab1A expression in lung cancer and paired noncancerous tissues

Article Snippet: Reagents were obtained from the following sources: antibody for Rab1A, Proteintech Group (Rosemont, IL); antibodies for P-S6K1(T398), S6K1, P-AKT(S473), AKT, P-ERK(T202/Y204), ERK, P-MK2(T334), MK2 and Tubulin, Cell Signaling Technology (Danvers, MA).

Techniques: Expressing

Journal: Aging (Albany NY)

Article Title: Expression of Rab1A is upregulated in human lung cancer and associated with tumor size and T stage

doi: 10.18632/aging.101087

Figure Lengend Snippet: Rab1A expression in five pathological subtypes of lung cancer

Article Snippet: Reagents were obtained from the following sources: antibody for Rab1A, Proteintech Group (Rosemont, IL); antibodies for P-S6K1(T398), S6K1, P-AKT(S473), AKT, P-ERK(T202/Y204), ERK, P-MK2(T334), MK2 and Tubulin, Cell Signaling Technology (Danvers, MA).

Techniques: Expressing

Journal: Aging (Albany NY)

Article Title: Expression of Rab1A is upregulated in human lung cancer and associated with tumor size and T stage

doi: 10.18632/aging.101087

Figure Lengend Snippet: Association between Rab1A expression and clinicopathologic features

Article Snippet: Reagents were obtained from the following sources: antibody for Rab1A, Proteintech Group (Rosemont, IL); antibodies for P-S6K1(T398), S6K1, P-AKT(S473), AKT, P-ERK(T202/Y204), ERK, P-MK2(T334), MK2 and Tubulin, Cell Signaling Technology (Danvers, MA).

Techniques: Expressing

Journal: Aging (Albany NY)

Article Title: Expression of Rab1A is upregulated in human lung cancer and associated with tumor size and T stage

doi: 10.18632/aging.101087

Figure Lengend Snippet: ( A ) Expression of Rab1A, P-S6K, S6K, P-AKT, AKT, P-ERK, ERK, P-MK2, MK2, P-c-Jun and c-Jun was examined in a panel of lung cancer cell lines by immunoblot. GAPDH served as a loading control. ( B ) Quantitative analysis of Rab1A protein levels in cell lines. The densitometry results for each Rab1A band were normalized to GAPDH, and the ratios between cancer cell lines and BEAS-2B were calculated and results represented in a histogram. Data represent means ± SD of three independent experiments. ( C-F ) Correlation plots of the expression level of Rab1A and P-S6K1 (T389; C ), P-AKT (S473; D ), P-ERK (T202/Y204; E ), P-MK2 (T334; F ) and P-c-Jun (S63; G ). Shown are arbitrary units.

Article Snippet: Reagents were obtained from the following sources: antibody for Rab1A, Proteintech Group (Rosemont, IL); antibodies for P-S6K1(T398), S6K1, P-AKT(S473), AKT, P-ERK(T202/Y204), ERK, P-MK2(T334), MK2 and Tubulin, Cell Signaling Technology (Danvers, MA).

Techniques: Expressing, Western Blot, Control

Journal: Aging (Albany NY)

Article Title: Expression of Rab1A is upregulated in human lung cancer and associated with tumor size and T stage

doi: 10.18632/aging.101087

Figure Lengend Snippet: ( A ) The effects of Rab1A knockdown was examined in H838, H1975, H650 and A549 cells by immunoblot. Levels of P-S6K1 (T389), S6K, P-AKT (S473) and AKT are shown. β-tubulin was used as a loading control. ( B ) Rab1A was knocked down in H838, H1975, H650 and A549 cells. The relative growth of these cells was analyzed using an MTT assay. Data represent means ± SD of three independent triplicate experiments. si-NC, negative control siRNA; si-Rab1A, Rab1A-specific siRNA.

Article Snippet: Reagents were obtained from the following sources: antibody for Rab1A, Proteintech Group (Rosemont, IL); antibodies for P-S6K1(T398), S6K1, P-AKT(S473), AKT, P-ERK(T202/Y204), ERK, P-MK2(T334), MK2 and Tubulin, Cell Signaling Technology (Danvers, MA).

Techniques: Knockdown, Western Blot, Control, MTT Assay, Negative Control