Review



anti mk2 rabbit polyclonal ab  (Cell Signaling Technology Inc)


Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 96
      Buy from Supplier

    Structured Review

    Cell Signaling Technology Inc anti mk2 rabbit polyclonal ab
    Activation of the p38 MAPK by various stress stimuli or growth factors results in activation of <t>MK2</t> and MK3. The signal transduction by these protein kinases towards the major substrates tristetraprolin (TTP) and HSPB1 is typically additive, although in most cells MK2 is the prevailing signal transducer, with little contribution of MK3. MK5 is activated by PKA, probably independently of p38 MAPK. The role of putative MK5 becomes obvious in the absence of MK2 and MK3, as shown in the present study. MK2, MK3, and MK5 phosphorylate the same two sites in mouse HSPB1 (Ser15, Ser86). In addition to MK2 and MK3, MNK1 and MSK1/2 are further MKs that are downstream of p38 MAPK. In macrophages, MK2 and MK3 were found to control expression of the immune response mediators IFNβ, IL-10, and NFκB through regulation of the activity of IRF3 and IκBβ. In these cells, MK2 was demonstrated to prevent MK3 from exerting negative regulatory effects on IRF3- and NFκB-dependent signaling. Dashed arrows indicate indirect signal transduction, while open arrows indicate complex effects on biological responses.
    Anti Mk2 Rabbit Polyclonal Ab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 606 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti mk2 rabbit polyclonal ab/product/Cell Signaling Technology Inc
    Average 96 stars, based on 606 article reviews
    anti mk2 rabbit polyclonal ab - by Bioz Stars, 2026-02
    96/100 stars

    Images

    1) Product Images from "Crucial Roles of the Protein Kinases MK2 and MK3 in a Mouse Model of Glomerulonephritis"

    Article Title: Crucial Roles of the Protein Kinases MK2 and MK3 in a Mouse Model of Glomerulonephritis

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0054239

    Activation of the p38 MAPK by various stress stimuli or growth factors results in activation of MK2 and MK3. The signal transduction by these protein kinases towards the major substrates tristetraprolin (TTP) and HSPB1 is typically additive, although in most cells MK2 is the prevailing signal transducer, with little contribution of MK3. MK5 is activated by PKA, probably independently of p38 MAPK. The role of putative MK5 becomes obvious in the absence of MK2 and MK3, as shown in the present study. MK2, MK3, and MK5 phosphorylate the same two sites in mouse HSPB1 (Ser15, Ser86). In addition to MK2 and MK3, MNK1 and MSK1/2 are further MKs that are downstream of p38 MAPK. In macrophages, MK2 and MK3 were found to control expression of the immune response mediators IFNβ, IL-10, and NFκB through regulation of the activity of IRF3 and IκBβ. In these cells, MK2 was demonstrated to prevent MK3 from exerting negative regulatory effects on IRF3- and NFκB-dependent signaling. Dashed arrows indicate indirect signal transduction, while open arrows indicate complex effects on biological responses.
    Figure Legend Snippet: Activation of the p38 MAPK by various stress stimuli or growth factors results in activation of MK2 and MK3. The signal transduction by these protein kinases towards the major substrates tristetraprolin (TTP) and HSPB1 is typically additive, although in most cells MK2 is the prevailing signal transducer, with little contribution of MK3. MK5 is activated by PKA, probably independently of p38 MAPK. The role of putative MK5 becomes obvious in the absence of MK2 and MK3, as shown in the present study. MK2, MK3, and MK5 phosphorylate the same two sites in mouse HSPB1 (Ser15, Ser86). In addition to MK2 and MK3, MNK1 and MSK1/2 are further MKs that are downstream of p38 MAPK. In macrophages, MK2 and MK3 were found to control expression of the immune response mediators IFNβ, IL-10, and NFκB through regulation of the activity of IRF3 and IκBβ. In these cells, MK2 was demonstrated to prevent MK3 from exerting negative regulatory effects on IRF3- and NFκB-dependent signaling. Dashed arrows indicate indirect signal transduction, while open arrows indicate complex effects on biological responses.

    Techniques Used: Activation Assay, Transduction, Control, Expressing, Activity Assay

    (A) Scatter plots show the urinary protein/creatinine ratios from samples collected from all mice at day 0 prior to AMC serum injection and at days 4, 8, and 12 following AMC serum injection. Horizontal bars indicate the means and error bars represent S.D. Asterisks indicate significant ( P <0.05) differences between means, as compared to the wild-type group at the same day. At days 8 and 12, all means were significantly greater than baseline proteinuria values at day 0 of the same genotypic group, with exception of the MK2/MK3 double knock-out ( MK2 −/− MK3 −/− ) mice at day 8. Given the high degree of variability within each experimental group, potential differences in mean proteinuria values among all other groups failed to reach statistical significance. Urine samples of mice selected for electrophoretic protein analysis as shown in (B) are labeled by numbers in the panels of days 4, 8, and 12. Note that in some instances the amount of collected urine was not sufficient for protein determination (i.e. the number of dots is less than the number of surviving mice as shown in ). (B) Urinary serum albumin excretion of selected mice (numbered 1 - 20) from the different MK2 and MK3 genotypes as visualized on Coomassie-stained SDS gels. Some of the selected mice survived throughout the entire experiment, while others died after day 4. The mouse numbers correspond to the numbered proteinuria values as indicated in (A). Consistent with the protein/creatinine ratios shown in (A), at day 0 mice of all genotypes had negligible albuminuria. Following AMC serum treatment, massive albuminuria was detected in most of the mice, with some variation in its extent and onset.
    Figure Legend Snippet: (A) Scatter plots show the urinary protein/creatinine ratios from samples collected from all mice at day 0 prior to AMC serum injection and at days 4, 8, and 12 following AMC serum injection. Horizontal bars indicate the means and error bars represent S.D. Asterisks indicate significant ( P <0.05) differences between means, as compared to the wild-type group at the same day. At days 8 and 12, all means were significantly greater than baseline proteinuria values at day 0 of the same genotypic group, with exception of the MK2/MK3 double knock-out ( MK2 −/− MK3 −/− ) mice at day 8. Given the high degree of variability within each experimental group, potential differences in mean proteinuria values among all other groups failed to reach statistical significance. Urine samples of mice selected for electrophoretic protein analysis as shown in (B) are labeled by numbers in the panels of days 4, 8, and 12. Note that in some instances the amount of collected urine was not sufficient for protein determination (i.e. the number of dots is less than the number of surviving mice as shown in ). (B) Urinary serum albumin excretion of selected mice (numbered 1 - 20) from the different MK2 and MK3 genotypes as visualized on Coomassie-stained SDS gels. Some of the selected mice survived throughout the entire experiment, while others died after day 4. The mouse numbers correspond to the numbered proteinuria values as indicated in (A). Consistent with the protein/creatinine ratios shown in (A), at day 0 mice of all genotypes had negligible albuminuria. Following AMC serum treatment, massive albuminuria was detected in most of the mice, with some variation in its extent and onset.

    Techniques Used: Injection, Knock-Out, Labeling, Staining

    Kidney sections from untreated and AMC serum-treated wild-type mice ( MK2 +/+ MK3 +/+ ), MK2/MK3 double knock-out mice ( MK2 −/− MK3 −/− ), and MK3 knock-out mice ( MK2 +/+ MK3 −/− ) on day 16 are shown. (A) Periodic acid-Schiff stain of the renal parenchyma of wild-type (panels a, c) and MK2/MK3 double knock-out mice (panels b, d), left untreated (panels a, b) or treated with the AMC serum (panels c, d). Normal morphology of the renal parenchyma was noted in untreated mice of both genotypes. Renal injury in response to the AMC serum included dilation of renal tubules and the presence of hyaline casts. Arrows designate glomeruli. Scale bar: 100 µM. (B) Silver stain (panels a–c, e) and trichrome stain (panels d, f) of glomeruli from untreated wild-type (panel a) and MK2/MK3 double knock-out mice (panel b), and from AMC serum-treated MK2/MK3 double knock-out (panels c–e) and MK3 knock-out mice (panel f). A preserved glomerular morphology was noted in untreated wild-type and MK2/MK3 double knock-out mice. Glomerular injury in response to the AMC serum included a thickening of the capillary walls due to duplication of basement membranes (tram-tracking) with associated mesangial interposition and narrowing of the capillary lumina (panel c, arrows), small fuchsinophilic subendothelial and mesangial deposits (panel d, arrows), and necrotizing lesions with associated crescent formation (panel e). These lesions were noted in mice of all genotypes. In addition, large, wire-loop type subendothelial deposits were found in the MK3 knock-out mice (panel f, arrows). Scale bar: 50 µM.
    Figure Legend Snippet: Kidney sections from untreated and AMC serum-treated wild-type mice ( MK2 +/+ MK3 +/+ ), MK2/MK3 double knock-out mice ( MK2 −/− MK3 −/− ), and MK3 knock-out mice ( MK2 +/+ MK3 −/− ) on day 16 are shown. (A) Periodic acid-Schiff stain of the renal parenchyma of wild-type (panels a, c) and MK2/MK3 double knock-out mice (panels b, d), left untreated (panels a, b) or treated with the AMC serum (panels c, d). Normal morphology of the renal parenchyma was noted in untreated mice of both genotypes. Renal injury in response to the AMC serum included dilation of renal tubules and the presence of hyaline casts. Arrows designate glomeruli. Scale bar: 100 µM. (B) Silver stain (panels a–c, e) and trichrome stain (panels d, f) of glomeruli from untreated wild-type (panel a) and MK2/MK3 double knock-out mice (panel b), and from AMC serum-treated MK2/MK3 double knock-out (panels c–e) and MK3 knock-out mice (panel f). A preserved glomerular morphology was noted in untreated wild-type and MK2/MK3 double knock-out mice. Glomerular injury in response to the AMC serum included a thickening of the capillary walls due to duplication of basement membranes (tram-tracking) with associated mesangial interposition and narrowing of the capillary lumina (panel c, arrows), small fuchsinophilic subendothelial and mesangial deposits (panel d, arrows), and necrotizing lesions with associated crescent formation (panel e). These lesions were noted in mice of all genotypes. In addition, large, wire-loop type subendothelial deposits were found in the MK3 knock-out mice (panel f, arrows). Scale bar: 50 µM.

    Techniques Used: Knock-Out, Staining, Silver Staining

    Extracts of renal cortices were processed for IEF-PAGE (A, panel a) or SDS-PAGE (A, panels b, c; B, C, D) from untreated mice (day 0; baseline control) and AMC serum-treated mice (day 8 of treatment). (A) Phosphorylation, baseline expression and induction of HSPB1. Panel a shows the distribution of the various HSPB1 isoforms (0p, unphosphorylated; 1p, singly phosphorylated; 2p, doubly phosphorylated) within each sample. Sample loading aimed to obtain comparable overall signals, in spite of considerable differences in the total HSPB1 content among the samples (cf. panel c). Panel b shows the amounts of Ser86-phosphorylated HSPB1 (p-Ser86). Equal amounts of total protein (15 µg) were loaded onto each lane. Panel c shows baseline expression and induction of HSPB1 in response to the AMC serum. (B) Baseline expression and response to the AMC serum of the heat shock proteins, HSPB6, HSPB8, and HSP70 (panels a–c, respectively). (C) Expression or phosphorylation of indicators of the unfolded protein response, CHOP (panel a), GRP78 (panel b), and eIF2α (panels c, d), before and after AMC serum treatment. Panels c and d show phosphorylated (p-eIF2α) and total eIF2α, respectively. (D) Expression of β-actin served as a loading control. Overall, this figure demonstrates partial involvement of MK2 and MK3 in baseline expression and/or phosphorylation of a number of sHSPs and indicators of the unfolded protein response, as well as in their pathophysiological response following AMC serum treatment.
    Figure Legend Snippet: Extracts of renal cortices were processed for IEF-PAGE (A, panel a) or SDS-PAGE (A, panels b, c; B, C, D) from untreated mice (day 0; baseline control) and AMC serum-treated mice (day 8 of treatment). (A) Phosphorylation, baseline expression and induction of HSPB1. Panel a shows the distribution of the various HSPB1 isoforms (0p, unphosphorylated; 1p, singly phosphorylated; 2p, doubly phosphorylated) within each sample. Sample loading aimed to obtain comparable overall signals, in spite of considerable differences in the total HSPB1 content among the samples (cf. panel c). Panel b shows the amounts of Ser86-phosphorylated HSPB1 (p-Ser86). Equal amounts of total protein (15 µg) were loaded onto each lane. Panel c shows baseline expression and induction of HSPB1 in response to the AMC serum. (B) Baseline expression and response to the AMC serum of the heat shock proteins, HSPB6, HSPB8, and HSP70 (panels a–c, respectively). (C) Expression or phosphorylation of indicators of the unfolded protein response, CHOP (panel a), GRP78 (panel b), and eIF2α (panels c, d), before and after AMC serum treatment. Panels c and d show phosphorylated (p-eIF2α) and total eIF2α, respectively. (D) Expression of β-actin served as a loading control. Overall, this figure demonstrates partial involvement of MK2 and MK3 in baseline expression and/or phosphorylation of a number of sHSPs and indicators of the unfolded protein response, as well as in their pathophysiological response following AMC serum treatment.

    Techniques Used: SDS Page, Control, Phospho-proteomics, Expressing

    Paraffin-embedded renal cortices of untreated and AMC serum-treated wild-type and MK2/MK3 double knock-out mice (day 16 following AMC serum treatment) were sectioned and processed for immunofluorescence microscopy. Total HSPB1 was visualized using an anti-HSPB1 antibody. In untreated control mice of either genotype, labeling of the glomeruli (including Bowman's space) was moderately elevated as compared to the surrounding tubules (upper row, left panels) or to the more distant tubules (upper row, right panels). AMC serum treatment caused a strong increase in HSPB1 labeling in the tubules, both adjacent to the glomeruli (lower row, left panels) and more distant from the glomeruli (lower row, right panels), thus indicating a stress response in the tubular compartment.
    Figure Legend Snippet: Paraffin-embedded renal cortices of untreated and AMC serum-treated wild-type and MK2/MK3 double knock-out mice (day 16 following AMC serum treatment) were sectioned and processed for immunofluorescence microscopy. Total HSPB1 was visualized using an anti-HSPB1 antibody. In untreated control mice of either genotype, labeling of the glomeruli (including Bowman's space) was moderately elevated as compared to the surrounding tubules (upper row, left panels) or to the more distant tubules (upper row, right panels). AMC serum treatment caused a strong increase in HSPB1 labeling in the tubules, both adjacent to the glomeruli (lower row, left panels) and more distant from the glomeruli (lower row, right panels), thus indicating a stress response in the tubular compartment.

    Techniques Used: Knock-Out, Immunofluorescence, Microscopy, Control, Labeling

    Summary of the observed baseline regulation in renal cortices with various MK2 and MK3 genotypes.
    Figure Legend Snippet: Summary of the observed baseline regulation in renal cortices with various MK2 and MK3 genotypes.

    Techniques Used: Phospho-proteomics

    Summary of the observed regulation in response to the AMC serum in renal cortices with various MK2 and MK3 genotypes.
    Figure Legend Snippet: Summary of the observed regulation in response to the AMC serum in renal cortices with various MK2 and MK3 genotypes.

    Techniques Used: Phospho-proteomics



    Similar Products

    anti mk2 rabbit polyclonal ab  (Cell Signaling Technology Inc)
    96
    Cell Signaling Technology Inc anti mk2 rabbit polyclonal ab
    Activation of the p38 MAPK by various stress stimuli or growth factors results in activation of <t>MK2</t> and MK3. The signal transduction by these protein kinases towards the major substrates tristetraprolin (TTP) and HSPB1 is typically additive, although in most cells MK2 is the prevailing signal transducer, with little contribution of MK3. MK5 is activated by PKA, probably independently of p38 MAPK. The role of putative MK5 becomes obvious in the absence of MK2 and MK3, as shown in the present study. MK2, MK3, and MK5 phosphorylate the same two sites in mouse HSPB1 (Ser15, Ser86). In addition to MK2 and MK3, MNK1 and MSK1/2 are further MKs that are downstream of p38 MAPK. In macrophages, MK2 and MK3 were found to control expression of the immune response mediators IFNβ, IL-10, and NFκB through regulation of the activity of IRF3 and IκBβ. In these cells, MK2 was demonstrated to prevent MK3 from exerting negative regulatory effects on IRF3- and NFκB-dependent signaling. Dashed arrows indicate indirect signal transduction, while open arrows indicate complex effects on biological responses.
    Anti Mk2 Rabbit Polyclonal Ab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti mk2 rabbit polyclonal ab/product/Cell Signaling Technology Inc
    Average 96 stars, based on 1 article reviews
    anti mk2 rabbit polyclonal ab - by Bioz Stars, 2026-02
    96/100 stars
    		  Buy from Supplier
    rabbit polyclonal anti mapkapk 2 abs  (Cell Signaling Technology Inc)
    96
    Cell Signaling Technology Inc rabbit polyclonal anti mapkapk 2 abs
    Activation of the p38 MAPK by various stress stimuli or growth factors results in activation of <t>MK2</t> and MK3. The signal transduction by these protein kinases towards the major substrates tristetraprolin (TTP) and HSPB1 is typically additive, although in most cells MK2 is the prevailing signal transducer, with little contribution of MK3. MK5 is activated by PKA, probably independently of p38 MAPK. The role of putative MK5 becomes obvious in the absence of MK2 and MK3, as shown in the present study. MK2, MK3, and MK5 phosphorylate the same two sites in mouse HSPB1 (Ser15, Ser86). In addition to MK2 and MK3, MNK1 and MSK1/2 are further MKs that are downstream of p38 MAPK. In macrophages, MK2 and MK3 were found to control expression of the immune response mediators IFNβ, IL-10, and NFκB through regulation of the activity of IRF3 and IκBβ. In these cells, MK2 was demonstrated to prevent MK3 from exerting negative regulatory effects on IRF3- and NFκB-dependent signaling. Dashed arrows indicate indirect signal transduction, while open arrows indicate complex effects on biological responses.
    Rabbit Polyclonal Anti Mapkapk 2 Abs, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal anti mapkapk 2 abs/product/Cell Signaling Technology Inc
    Average 96 stars, based on 1 article reviews
    rabbit polyclonal anti mapkapk 2 abs - by Bioz Stars, 2026-02
    96/100 stars
    		  Buy from Supplier
    rabbit polyclonal anti mapkapk 2 abs  (Bio-Rad)
    85
    Bio-Rad rabbit polyclonal anti mapkapk 2 abs
    Activation of the p38 MAPK by various stress stimuli or growth factors results in activation of <t>MK2</t> and MK3. The signal transduction by these protein kinases towards the major substrates tristetraprolin (TTP) and HSPB1 is typically additive, although in most cells MK2 is the prevailing signal transducer, with little contribution of MK3. MK5 is activated by PKA, probably independently of p38 MAPK. The role of putative MK5 becomes obvious in the absence of MK2 and MK3, as shown in the present study. MK2, MK3, and MK5 phosphorylate the same two sites in mouse HSPB1 (Ser15, Ser86). In addition to MK2 and MK3, MNK1 and MSK1/2 are further MKs that are downstream of p38 MAPK. In macrophages, MK2 and MK3 were found to control expression of the immune response mediators IFNβ, IL-10, and NFκB through regulation of the activity of IRF3 and IκBβ. In these cells, MK2 was demonstrated to prevent MK3 from exerting negative regulatory effects on IRF3- and NFκB-dependent signaling. Dashed arrows indicate indirect signal transduction, while open arrows indicate complex effects on biological responses.
    Rabbit Polyclonal Anti Mapkapk 2 Abs, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal anti mapkapk 2 abs/product/Bio-Rad
    Average 85 stars, based on 1 article reviews
    rabbit polyclonal anti mapkapk 2 abs - by Bioz Stars, 2026-02
    85/100 stars
    		  Buy from Supplier

    Image Search Results


    Activation of the p38 MAPK by various stress stimuli or growth factors results in activation of MK2 and MK3. The signal transduction by these protein kinases towards the major substrates tristetraprolin (TTP) and HSPB1 is typically additive, although in most cells MK2 is the prevailing signal transducer, with little contribution of MK3. MK5 is activated by PKA, probably independently of p38 MAPK. The role of putative MK5 becomes obvious in the absence of MK2 and MK3, as shown in the present study. MK2, MK3, and MK5 phosphorylate the same two sites in mouse HSPB1 (Ser15, Ser86). In addition to MK2 and MK3, MNK1 and MSK1/2 are further MKs that are downstream of p38 MAPK. In macrophages, MK2 and MK3 were found to control expression of the immune response mediators IFNβ, IL-10, and NFκB through regulation of the activity of IRF3 and IκBβ. In these cells, MK2 was demonstrated to prevent MK3 from exerting negative regulatory effects on IRF3- and NFκB-dependent signaling. Dashed arrows indicate indirect signal transduction, while open arrows indicate complex effects on biological responses.

    Journal: PLoS ONE

    Article Title: Crucial Roles of the Protein Kinases MK2 and MK3 in a Mouse Model of Glomerulonephritis

    doi: 10.1371/journal.pone.0054239

    Figure Lengend Snippet: Activation of the p38 MAPK by various stress stimuli or growth factors results in activation of MK2 and MK3. The signal transduction by these protein kinases towards the major substrates tristetraprolin (TTP) and HSPB1 is typically additive, although in most cells MK2 is the prevailing signal transducer, with little contribution of MK3. MK5 is activated by PKA, probably independently of p38 MAPK. The role of putative MK5 becomes obvious in the absence of MK2 and MK3, as shown in the present study. MK2, MK3, and MK5 phosphorylate the same two sites in mouse HSPB1 (Ser15, Ser86). In addition to MK2 and MK3, MNK1 and MSK1/2 are further MKs that are downstream of p38 MAPK. In macrophages, MK2 and MK3 were found to control expression of the immune response mediators IFNβ, IL-10, and NFκB through regulation of the activity of IRF3 and IκBβ. In these cells, MK2 was demonstrated to prevent MK3 from exerting negative regulatory effects on IRF3- and NFκB-dependent signaling. Dashed arrows indicate indirect signal transduction, while open arrows indicate complex effects on biological responses.

    Article Snippet: The following primary antibodies (Ab) were used: anti-MK2 rabbit polyclonal Ab (dilution: 1∶1000; Cell Signaling, Danvers, MA), anti-MK3 rabbit monoclonal Ab (dilution 1∶1000; Cell Signaling), anti-β-actin rabbit monoclonal Ab (dilution: 1∶5000; Cell Signaling), anti-eIF2α mouse monoclonal Ab (dilution: 1∶1000; Cell Signaling), anti-phospho-eIF2α rabbit monoclonal Ab (dilution: 1∶1000; Cell Signaling), anti-CHOP mouse monoclonal Ab (dilution: 1∶1000; Cell Signaling), anti-HSPB1 rabbit polyclonal Ab (dilution: 1∶5000; Assay Designs), anti-phospho-HSPB1-Ser86 rabbit polyclonal Ab (dilution: 1∶1000; Cell Signaling), anti-GRP78 rabbit polyclonal Ab (dilution: 1∶1000; StressMarq, Victoria, BC), anti-Hsp70 mouse monoclonal Ab (dilution: 1∶1000; StressMarq), anti-HSPB8 mouse monoclonal Ab (dilution: 1∶1000; Abcam, Cambridge, MA), anti-HSPB6 rabbit polyclonal Ab (dilution: 1∶1000; Abcam), anti-MK5 rabbit monoclonal Ab (dilution 1∶1000; Cell Signaling), anti-phospho-MK5 rabbit polyclonal Ab (dilution 1∶500; Pierce Biotechnology, Rockford, IL), anti-MNK1 rabbit monoclonal Ab (dilution 1∶1000; Cell Signaling), anti-phospho-Mnk1 polyclonal Ab (dilution 1∶1000; Cell Signaling), anti-MSK1 rabbit polyclonal Ab (dilution 1∶500; LifeSpan BioSciences, Seattle, WA), anti-phospho-MSK1 rabbit polyclonal Ab (dilution 1∶500; R&D Systems, Minneapolis, MN), and anti-GAPDH mouse monoclonal Ab (dilution 1∶5000; Millipore, Billerica, MA).

    Techniques: Activation Assay, Transduction, Control, Expressing, Activity Assay

    (A) Scatter plots show the urinary protein/creatinine ratios from samples collected from all mice at day 0 prior to AMC serum injection and at days 4, 8, and 12 following AMC serum injection. Horizontal bars indicate the means and error bars represent S.D. Asterisks indicate significant ( P <0.05) differences between means, as compared to the wild-type group at the same day. At days 8 and 12, all means were significantly greater than baseline proteinuria values at day 0 of the same genotypic group, with exception of the MK2/MK3 double knock-out ( MK2 −/− MK3 −/− ) mice at day 8. Given the high degree of variability within each experimental group, potential differences in mean proteinuria values among all other groups failed to reach statistical significance. Urine samples of mice selected for electrophoretic protein analysis as shown in (B) are labeled by numbers in the panels of days 4, 8, and 12. Note that in some instances the amount of collected urine was not sufficient for protein determination (i.e. the number of dots is less than the number of surviving mice as shown in ). (B) Urinary serum albumin excretion of selected mice (numbered 1 - 20) from the different MK2 and MK3 genotypes as visualized on Coomassie-stained SDS gels. Some of the selected mice survived throughout the entire experiment, while others died after day 4. The mouse numbers correspond to the numbered proteinuria values as indicated in (A). Consistent with the protein/creatinine ratios shown in (A), at day 0 mice of all genotypes had negligible albuminuria. Following AMC serum treatment, massive albuminuria was detected in most of the mice, with some variation in its extent and onset.

    Journal: PLoS ONE

    Article Title: Crucial Roles of the Protein Kinases MK2 and MK3 in a Mouse Model of Glomerulonephritis

    doi: 10.1371/journal.pone.0054239

    Figure Lengend Snippet: (A) Scatter plots show the urinary protein/creatinine ratios from samples collected from all mice at day 0 prior to AMC serum injection and at days 4, 8, and 12 following AMC serum injection. Horizontal bars indicate the means and error bars represent S.D. Asterisks indicate significant ( P <0.05) differences between means, as compared to the wild-type group at the same day. At days 8 and 12, all means were significantly greater than baseline proteinuria values at day 0 of the same genotypic group, with exception of the MK2/MK3 double knock-out ( MK2 −/− MK3 −/− ) mice at day 8. Given the high degree of variability within each experimental group, potential differences in mean proteinuria values among all other groups failed to reach statistical significance. Urine samples of mice selected for electrophoretic protein analysis as shown in (B) are labeled by numbers in the panels of days 4, 8, and 12. Note that in some instances the amount of collected urine was not sufficient for protein determination (i.e. the number of dots is less than the number of surviving mice as shown in ). (B) Urinary serum albumin excretion of selected mice (numbered 1 - 20) from the different MK2 and MK3 genotypes as visualized on Coomassie-stained SDS gels. Some of the selected mice survived throughout the entire experiment, while others died after day 4. The mouse numbers correspond to the numbered proteinuria values as indicated in (A). Consistent with the protein/creatinine ratios shown in (A), at day 0 mice of all genotypes had negligible albuminuria. Following AMC serum treatment, massive albuminuria was detected in most of the mice, with some variation in its extent and onset.

    Article Snippet: The following primary antibodies (Ab) were used: anti-MK2 rabbit polyclonal Ab (dilution: 1∶1000; Cell Signaling, Danvers, MA), anti-MK3 rabbit monoclonal Ab (dilution 1∶1000; Cell Signaling), anti-β-actin rabbit monoclonal Ab (dilution: 1∶5000; Cell Signaling), anti-eIF2α mouse monoclonal Ab (dilution: 1∶1000; Cell Signaling), anti-phospho-eIF2α rabbit monoclonal Ab (dilution: 1∶1000; Cell Signaling), anti-CHOP mouse monoclonal Ab (dilution: 1∶1000; Cell Signaling), anti-HSPB1 rabbit polyclonal Ab (dilution: 1∶5000; Assay Designs), anti-phospho-HSPB1-Ser86 rabbit polyclonal Ab (dilution: 1∶1000; Cell Signaling), anti-GRP78 rabbit polyclonal Ab (dilution: 1∶1000; StressMarq, Victoria, BC), anti-Hsp70 mouse monoclonal Ab (dilution: 1∶1000; StressMarq), anti-HSPB8 mouse monoclonal Ab (dilution: 1∶1000; Abcam, Cambridge, MA), anti-HSPB6 rabbit polyclonal Ab (dilution: 1∶1000; Abcam), anti-MK5 rabbit monoclonal Ab (dilution 1∶1000; Cell Signaling), anti-phospho-MK5 rabbit polyclonal Ab (dilution 1∶500; Pierce Biotechnology, Rockford, IL), anti-MNK1 rabbit monoclonal Ab (dilution 1∶1000; Cell Signaling), anti-phospho-Mnk1 polyclonal Ab (dilution 1∶1000; Cell Signaling), anti-MSK1 rabbit polyclonal Ab (dilution 1∶500; LifeSpan BioSciences, Seattle, WA), anti-phospho-MSK1 rabbit polyclonal Ab (dilution 1∶500; R&D Systems, Minneapolis, MN), and anti-GAPDH mouse monoclonal Ab (dilution 1∶5000; Millipore, Billerica, MA).

    Techniques: Injection, Knock-Out, Labeling, Staining

    Kidney sections from untreated and AMC serum-treated wild-type mice ( MK2 +/+ MK3 +/+ ), MK2/MK3 double knock-out mice ( MK2 −/− MK3 −/− ), and MK3 knock-out mice ( MK2 +/+ MK3 −/− ) on day 16 are shown. (A) Periodic acid-Schiff stain of the renal parenchyma of wild-type (panels a, c) and MK2/MK3 double knock-out mice (panels b, d), left untreated (panels a, b) or treated with the AMC serum (panels c, d). Normal morphology of the renal parenchyma was noted in untreated mice of both genotypes. Renal injury in response to the AMC serum included dilation of renal tubules and the presence of hyaline casts. Arrows designate glomeruli. Scale bar: 100 µM. (B) Silver stain (panels a–c, e) and trichrome stain (panels d, f) of glomeruli from untreated wild-type (panel a) and MK2/MK3 double knock-out mice (panel b), and from AMC serum-treated MK2/MK3 double knock-out (panels c–e) and MK3 knock-out mice (panel f). A preserved glomerular morphology was noted in untreated wild-type and MK2/MK3 double knock-out mice. Glomerular injury in response to the AMC serum included a thickening of the capillary walls due to duplication of basement membranes (tram-tracking) with associated mesangial interposition and narrowing of the capillary lumina (panel c, arrows), small fuchsinophilic subendothelial and mesangial deposits (panel d, arrows), and necrotizing lesions with associated crescent formation (panel e). These lesions were noted in mice of all genotypes. In addition, large, wire-loop type subendothelial deposits were found in the MK3 knock-out mice (panel f, arrows). Scale bar: 50 µM.

    Journal: PLoS ONE

    Article Title: Crucial Roles of the Protein Kinases MK2 and MK3 in a Mouse Model of Glomerulonephritis

    doi: 10.1371/journal.pone.0054239

    Figure Lengend Snippet: Kidney sections from untreated and AMC serum-treated wild-type mice ( MK2 +/+ MK3 +/+ ), MK2/MK3 double knock-out mice ( MK2 −/− MK3 −/− ), and MK3 knock-out mice ( MK2 +/+ MK3 −/− ) on day 16 are shown. (A) Periodic acid-Schiff stain of the renal parenchyma of wild-type (panels a, c) and MK2/MK3 double knock-out mice (panels b, d), left untreated (panels a, b) or treated with the AMC serum (panels c, d). Normal morphology of the renal parenchyma was noted in untreated mice of both genotypes. Renal injury in response to the AMC serum included dilation of renal tubules and the presence of hyaline casts. Arrows designate glomeruli. Scale bar: 100 µM. (B) Silver stain (panels a–c, e) and trichrome stain (panels d, f) of glomeruli from untreated wild-type (panel a) and MK2/MK3 double knock-out mice (panel b), and from AMC serum-treated MK2/MK3 double knock-out (panels c–e) and MK3 knock-out mice (panel f). A preserved glomerular morphology was noted in untreated wild-type and MK2/MK3 double knock-out mice. Glomerular injury in response to the AMC serum included a thickening of the capillary walls due to duplication of basement membranes (tram-tracking) with associated mesangial interposition and narrowing of the capillary lumina (panel c, arrows), small fuchsinophilic subendothelial and mesangial deposits (panel d, arrows), and necrotizing lesions with associated crescent formation (panel e). These lesions were noted in mice of all genotypes. In addition, large, wire-loop type subendothelial deposits were found in the MK3 knock-out mice (panel f, arrows). Scale bar: 50 µM.

    Article Snippet: The following primary antibodies (Ab) were used: anti-MK2 rabbit polyclonal Ab (dilution: 1∶1000; Cell Signaling, Danvers, MA), anti-MK3 rabbit monoclonal Ab (dilution 1∶1000; Cell Signaling), anti-β-actin rabbit monoclonal Ab (dilution: 1∶5000; Cell Signaling), anti-eIF2α mouse monoclonal Ab (dilution: 1∶1000; Cell Signaling), anti-phospho-eIF2α rabbit monoclonal Ab (dilution: 1∶1000; Cell Signaling), anti-CHOP mouse monoclonal Ab (dilution: 1∶1000; Cell Signaling), anti-HSPB1 rabbit polyclonal Ab (dilution: 1∶5000; Assay Designs), anti-phospho-HSPB1-Ser86 rabbit polyclonal Ab (dilution: 1∶1000; Cell Signaling), anti-GRP78 rabbit polyclonal Ab (dilution: 1∶1000; StressMarq, Victoria, BC), anti-Hsp70 mouse monoclonal Ab (dilution: 1∶1000; StressMarq), anti-HSPB8 mouse monoclonal Ab (dilution: 1∶1000; Abcam, Cambridge, MA), anti-HSPB6 rabbit polyclonal Ab (dilution: 1∶1000; Abcam), anti-MK5 rabbit monoclonal Ab (dilution 1∶1000; Cell Signaling), anti-phospho-MK5 rabbit polyclonal Ab (dilution 1∶500; Pierce Biotechnology, Rockford, IL), anti-MNK1 rabbit monoclonal Ab (dilution 1∶1000; Cell Signaling), anti-phospho-Mnk1 polyclonal Ab (dilution 1∶1000; Cell Signaling), anti-MSK1 rabbit polyclonal Ab (dilution 1∶500; LifeSpan BioSciences, Seattle, WA), anti-phospho-MSK1 rabbit polyclonal Ab (dilution 1∶500; R&D Systems, Minneapolis, MN), and anti-GAPDH mouse monoclonal Ab (dilution 1∶5000; Millipore, Billerica, MA).

    Techniques: Knock-Out, Staining, Silver Staining

    Extracts of renal cortices were processed for IEF-PAGE (A, panel a) or SDS-PAGE (A, panels b, c; B, C, D) from untreated mice (day 0; baseline control) and AMC serum-treated mice (day 8 of treatment). (A) Phosphorylation, baseline expression and induction of HSPB1. Panel a shows the distribution of the various HSPB1 isoforms (0p, unphosphorylated; 1p, singly phosphorylated; 2p, doubly phosphorylated) within each sample. Sample loading aimed to obtain comparable overall signals, in spite of considerable differences in the total HSPB1 content among the samples (cf. panel c). Panel b shows the amounts of Ser86-phosphorylated HSPB1 (p-Ser86). Equal amounts of total protein (15 µg) were loaded onto each lane. Panel c shows baseline expression and induction of HSPB1 in response to the AMC serum. (B) Baseline expression and response to the AMC serum of the heat shock proteins, HSPB6, HSPB8, and HSP70 (panels a–c, respectively). (C) Expression or phosphorylation of indicators of the unfolded protein response, CHOP (panel a), GRP78 (panel b), and eIF2α (panels c, d), before and after AMC serum treatment. Panels c and d show phosphorylated (p-eIF2α) and total eIF2α, respectively. (D) Expression of β-actin served as a loading control. Overall, this figure demonstrates partial involvement of MK2 and MK3 in baseline expression and/or phosphorylation of a number of sHSPs and indicators of the unfolded protein response, as well as in their pathophysiological response following AMC serum treatment.

    Journal: PLoS ONE

    Article Title: Crucial Roles of the Protein Kinases MK2 and MK3 in a Mouse Model of Glomerulonephritis

    doi: 10.1371/journal.pone.0054239

    Figure Lengend Snippet: Extracts of renal cortices were processed for IEF-PAGE (A, panel a) or SDS-PAGE (A, panels b, c; B, C, D) from untreated mice (day 0; baseline control) and AMC serum-treated mice (day 8 of treatment). (A) Phosphorylation, baseline expression and induction of HSPB1. Panel a shows the distribution of the various HSPB1 isoforms (0p, unphosphorylated; 1p, singly phosphorylated; 2p, doubly phosphorylated) within each sample. Sample loading aimed to obtain comparable overall signals, in spite of considerable differences in the total HSPB1 content among the samples (cf. panel c). Panel b shows the amounts of Ser86-phosphorylated HSPB1 (p-Ser86). Equal amounts of total protein (15 µg) were loaded onto each lane. Panel c shows baseline expression and induction of HSPB1 in response to the AMC serum. (B) Baseline expression and response to the AMC serum of the heat shock proteins, HSPB6, HSPB8, and HSP70 (panels a–c, respectively). (C) Expression or phosphorylation of indicators of the unfolded protein response, CHOP (panel a), GRP78 (panel b), and eIF2α (panels c, d), before and after AMC serum treatment. Panels c and d show phosphorylated (p-eIF2α) and total eIF2α, respectively. (D) Expression of β-actin served as a loading control. Overall, this figure demonstrates partial involvement of MK2 and MK3 in baseline expression and/or phosphorylation of a number of sHSPs and indicators of the unfolded protein response, as well as in their pathophysiological response following AMC serum treatment.

    Article Snippet: The following primary antibodies (Ab) were used: anti-MK2 rabbit polyclonal Ab (dilution: 1∶1000; Cell Signaling, Danvers, MA), anti-MK3 rabbit monoclonal Ab (dilution 1∶1000; Cell Signaling), anti-β-actin rabbit monoclonal Ab (dilution: 1∶5000; Cell Signaling), anti-eIF2α mouse monoclonal Ab (dilution: 1∶1000; Cell Signaling), anti-phospho-eIF2α rabbit monoclonal Ab (dilution: 1∶1000; Cell Signaling), anti-CHOP mouse monoclonal Ab (dilution: 1∶1000; Cell Signaling), anti-HSPB1 rabbit polyclonal Ab (dilution: 1∶5000; Assay Designs), anti-phospho-HSPB1-Ser86 rabbit polyclonal Ab (dilution: 1∶1000; Cell Signaling), anti-GRP78 rabbit polyclonal Ab (dilution: 1∶1000; StressMarq, Victoria, BC), anti-Hsp70 mouse monoclonal Ab (dilution: 1∶1000; StressMarq), anti-HSPB8 mouse monoclonal Ab (dilution: 1∶1000; Abcam, Cambridge, MA), anti-HSPB6 rabbit polyclonal Ab (dilution: 1∶1000; Abcam), anti-MK5 rabbit monoclonal Ab (dilution 1∶1000; Cell Signaling), anti-phospho-MK5 rabbit polyclonal Ab (dilution 1∶500; Pierce Biotechnology, Rockford, IL), anti-MNK1 rabbit monoclonal Ab (dilution 1∶1000; Cell Signaling), anti-phospho-Mnk1 polyclonal Ab (dilution 1∶1000; Cell Signaling), anti-MSK1 rabbit polyclonal Ab (dilution 1∶500; LifeSpan BioSciences, Seattle, WA), anti-phospho-MSK1 rabbit polyclonal Ab (dilution 1∶500; R&D Systems, Minneapolis, MN), and anti-GAPDH mouse monoclonal Ab (dilution 1∶5000; Millipore, Billerica, MA).

    Techniques: SDS Page, Control, Phospho-proteomics, Expressing

    Paraffin-embedded renal cortices of untreated and AMC serum-treated wild-type and MK2/MK3 double knock-out mice (day 16 following AMC serum treatment) were sectioned and processed for immunofluorescence microscopy. Total HSPB1 was visualized using an anti-HSPB1 antibody. In untreated control mice of either genotype, labeling of the glomeruli (including Bowman's space) was moderately elevated as compared to the surrounding tubules (upper row, left panels) or to the more distant tubules (upper row, right panels). AMC serum treatment caused a strong increase in HSPB1 labeling in the tubules, both adjacent to the glomeruli (lower row, left panels) and more distant from the glomeruli (lower row, right panels), thus indicating a stress response in the tubular compartment.

    Journal: PLoS ONE

    Article Title: Crucial Roles of the Protein Kinases MK2 and MK3 in a Mouse Model of Glomerulonephritis

    doi: 10.1371/journal.pone.0054239

    Figure Lengend Snippet: Paraffin-embedded renal cortices of untreated and AMC serum-treated wild-type and MK2/MK3 double knock-out mice (day 16 following AMC serum treatment) were sectioned and processed for immunofluorescence microscopy. Total HSPB1 was visualized using an anti-HSPB1 antibody. In untreated control mice of either genotype, labeling of the glomeruli (including Bowman's space) was moderately elevated as compared to the surrounding tubules (upper row, left panels) or to the more distant tubules (upper row, right panels). AMC serum treatment caused a strong increase in HSPB1 labeling in the tubules, both adjacent to the glomeruli (lower row, left panels) and more distant from the glomeruli (lower row, right panels), thus indicating a stress response in the tubular compartment.

    Article Snippet: The following primary antibodies (Ab) were used: anti-MK2 rabbit polyclonal Ab (dilution: 1∶1000; Cell Signaling, Danvers, MA), anti-MK3 rabbit monoclonal Ab (dilution 1∶1000; Cell Signaling), anti-β-actin rabbit monoclonal Ab (dilution: 1∶5000; Cell Signaling), anti-eIF2α mouse monoclonal Ab (dilution: 1∶1000; Cell Signaling), anti-phospho-eIF2α rabbit monoclonal Ab (dilution: 1∶1000; Cell Signaling), anti-CHOP mouse monoclonal Ab (dilution: 1∶1000; Cell Signaling), anti-HSPB1 rabbit polyclonal Ab (dilution: 1∶5000; Assay Designs), anti-phospho-HSPB1-Ser86 rabbit polyclonal Ab (dilution: 1∶1000; Cell Signaling), anti-GRP78 rabbit polyclonal Ab (dilution: 1∶1000; StressMarq, Victoria, BC), anti-Hsp70 mouse monoclonal Ab (dilution: 1∶1000; StressMarq), anti-HSPB8 mouse monoclonal Ab (dilution: 1∶1000; Abcam, Cambridge, MA), anti-HSPB6 rabbit polyclonal Ab (dilution: 1∶1000; Abcam), anti-MK5 rabbit monoclonal Ab (dilution 1∶1000; Cell Signaling), anti-phospho-MK5 rabbit polyclonal Ab (dilution 1∶500; Pierce Biotechnology, Rockford, IL), anti-MNK1 rabbit monoclonal Ab (dilution 1∶1000; Cell Signaling), anti-phospho-Mnk1 polyclonal Ab (dilution 1∶1000; Cell Signaling), anti-MSK1 rabbit polyclonal Ab (dilution 1∶500; LifeSpan BioSciences, Seattle, WA), anti-phospho-MSK1 rabbit polyclonal Ab (dilution 1∶500; R&D Systems, Minneapolis, MN), and anti-GAPDH mouse monoclonal Ab (dilution 1∶5000; Millipore, Billerica, MA).

    Techniques: Knock-Out, Immunofluorescence, Microscopy, Control, Labeling

    Summary of the observed baseline regulation in renal cortices with various MK2 and MK3 genotypes.

    Journal: PLoS ONE

    Article Title: Crucial Roles of the Protein Kinases MK2 and MK3 in a Mouse Model of Glomerulonephritis

    doi: 10.1371/journal.pone.0054239

    Figure Lengend Snippet: Summary of the observed baseline regulation in renal cortices with various MK2 and MK3 genotypes.

    Article Snippet: The following primary antibodies (Ab) were used: anti-MK2 rabbit polyclonal Ab (dilution: 1∶1000; Cell Signaling, Danvers, MA), anti-MK3 rabbit monoclonal Ab (dilution 1∶1000; Cell Signaling), anti-β-actin rabbit monoclonal Ab (dilution: 1∶5000; Cell Signaling), anti-eIF2α mouse monoclonal Ab (dilution: 1∶1000; Cell Signaling), anti-phospho-eIF2α rabbit monoclonal Ab (dilution: 1∶1000; Cell Signaling), anti-CHOP mouse monoclonal Ab (dilution: 1∶1000; Cell Signaling), anti-HSPB1 rabbit polyclonal Ab (dilution: 1∶5000; Assay Designs), anti-phospho-HSPB1-Ser86 rabbit polyclonal Ab (dilution: 1∶1000; Cell Signaling), anti-GRP78 rabbit polyclonal Ab (dilution: 1∶1000; StressMarq, Victoria, BC), anti-Hsp70 mouse monoclonal Ab (dilution: 1∶1000; StressMarq), anti-HSPB8 mouse monoclonal Ab (dilution: 1∶1000; Abcam, Cambridge, MA), anti-HSPB6 rabbit polyclonal Ab (dilution: 1∶1000; Abcam), anti-MK5 rabbit monoclonal Ab (dilution 1∶1000; Cell Signaling), anti-phospho-MK5 rabbit polyclonal Ab (dilution 1∶500; Pierce Biotechnology, Rockford, IL), anti-MNK1 rabbit monoclonal Ab (dilution 1∶1000; Cell Signaling), anti-phospho-Mnk1 polyclonal Ab (dilution 1∶1000; Cell Signaling), anti-MSK1 rabbit polyclonal Ab (dilution 1∶500; LifeSpan BioSciences, Seattle, WA), anti-phospho-MSK1 rabbit polyclonal Ab (dilution 1∶500; R&D Systems, Minneapolis, MN), and anti-GAPDH mouse monoclonal Ab (dilution 1∶5000; Millipore, Billerica, MA).

    Techniques: Phospho-proteomics

    Summary of the observed regulation in response to the AMC serum in renal cortices with various MK2 and MK3 genotypes.

    Journal: PLoS ONE

    Article Title: Crucial Roles of the Protein Kinases MK2 and MK3 in a Mouse Model of Glomerulonephritis

    doi: 10.1371/journal.pone.0054239

    Figure Lengend Snippet: Summary of the observed regulation in response to the AMC serum in renal cortices with various MK2 and MK3 genotypes.

    Article Snippet: The following primary antibodies (Ab) were used: anti-MK2 rabbit polyclonal Ab (dilution: 1∶1000; Cell Signaling, Danvers, MA), anti-MK3 rabbit monoclonal Ab (dilution 1∶1000; Cell Signaling), anti-β-actin rabbit monoclonal Ab (dilution: 1∶5000; Cell Signaling), anti-eIF2α mouse monoclonal Ab (dilution: 1∶1000; Cell Signaling), anti-phospho-eIF2α rabbit monoclonal Ab (dilution: 1∶1000; Cell Signaling), anti-CHOP mouse monoclonal Ab (dilution: 1∶1000; Cell Signaling), anti-HSPB1 rabbit polyclonal Ab (dilution: 1∶5000; Assay Designs), anti-phospho-HSPB1-Ser86 rabbit polyclonal Ab (dilution: 1∶1000; Cell Signaling), anti-GRP78 rabbit polyclonal Ab (dilution: 1∶1000; StressMarq, Victoria, BC), anti-Hsp70 mouse monoclonal Ab (dilution: 1∶1000; StressMarq), anti-HSPB8 mouse monoclonal Ab (dilution: 1∶1000; Abcam, Cambridge, MA), anti-HSPB6 rabbit polyclonal Ab (dilution: 1∶1000; Abcam), anti-MK5 rabbit monoclonal Ab (dilution 1∶1000; Cell Signaling), anti-phospho-MK5 rabbit polyclonal Ab (dilution 1∶500; Pierce Biotechnology, Rockford, IL), anti-MNK1 rabbit monoclonal Ab (dilution 1∶1000; Cell Signaling), anti-phospho-Mnk1 polyclonal Ab (dilution 1∶1000; Cell Signaling), anti-MSK1 rabbit polyclonal Ab (dilution 1∶500; LifeSpan BioSciences, Seattle, WA), anti-phospho-MSK1 rabbit polyclonal Ab (dilution 1∶500; R&D Systems, Minneapolis, MN), and anti-GAPDH mouse monoclonal Ab (dilution 1∶5000; Millipore, Billerica, MA).

    Techniques: Phospho-proteomics