Journal: International journal of molecular medicine

Article Title: Rhododendron album Blume inhibits iNOS and COX-2 expression in LPS-stimulated RAW264.7 cells through the downregulation of NF-κB signaling.

doi: 10.3892/ijmm.2015.2107

Figure Lengend Snippet: Figure 4. Rhododendron album Blume methanol extract (RAME) suppresses the phosphorylation of mitogen-activated protein kinase (MAPK)‑associated molecules in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Cells were pre-incubated with RAME for 12 h and then incubated with LPS (0.5 µg/ml) for 20 min. The total protein lysate was subjected to western blot analysis. The ratio of the immunointensity of p-ERK1/2, p-JNK and p-p38 was calculated. Total ERK1/2, JNK and p38 (ERK1/2, JNK and p38) were used as a control for the amount of protein in the same samples. A representative blot of each experiment is shown with the densitometric analysis corresponding to the means ± SEM of 3 independent experiments.

Article Snippet: Each membrane was then incubated for 1 h in 5% skim milk in TBS-T buffer (0.1 M Tris-HCl, pH 7.4, 0.9% NaCl, 0.1% Tween-20) to block non-specific binding and was then incubated with primary antibodies that recognized iNOS (Cat. no. ADI-905-431, 1:1,000; obtained from Enzo Life Sciences, Farmingdale, NY, USA), COX-2 (Cat. no. sc-1747, 1:1,000; Santa Cruz Biotechnology, Santa Cruz, CA, USA), β-actin (Cat. no. #4967, 1:2,000; Cell Signaling Technology, Danvers, MA, USA), PARP (Cat. no. #9542; Cell Signaling Technology), the total forms of extracellular signal-regulated kinase (ERK)2 (sc-154), p38 MAPK (sc-7149), c-Jun N-terminal kinase (JNK)1/3 (sc-474, 1:1,000; Santa Cruz Biotechnology), the phosphorylated forms of p38 MAPK (ADI-KAP-MA022) and JNK1/2 (KAP-SA011, 1:1,000; Enzo Life Sciences, Inc.) and the phosphorylated forms of ERK (#9106, 1:1,000; Cell Signaling Technology).

Techniques: Phospho-proteomics, Incubation, Western Blot, Control

Journal: Experimental and Therapeutic Medicine

Article Title: Fistular onion stalk extract exhibits anti-atherosclerotic effects in rats

doi: 10.3892/etm.2014.1790

Figure Lengend Snippet: Fistular onion stalk extract inhibits local inflammatory signaling pathways. The control group maintained a low expression and the model group had a significantly upregulated expression of p-p65, p-stat3 and p-p38. However, the p-p65, p-stat3 and p-p38 expression in the treatment group was downregulated and maintained at a lower level than that in the model group. Total expression levels of p65, stat3 and p38 were similar among the different groups. GAPDH expression was set as an internal control. p-, phosphorylated-; stat3, signal transducer and activator of transcription 3.

Article Snippet: The following antibodies were used at a dilution of 1:1,000 : Anti-phosphorylated-(p-)p65 (sc-101749), anti-p65 (sc-8008), anti-p-stat3 (sc-135649), anti-stat3 (sc-8019), anti-p-p38 (sc-7973) and anti-p38 (sc-7149) (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA).

Techniques: Protein-Protein interactions, Control, Expressing

Journal: Molecular medicine reports

Article Title: Inhibitory effect of Zanthoxylum bungeanum seed oil on ovalbumin‑induced lung inflammation in a murine model of asthma.

doi: 10.3892/mmr.2016.5050

Figure Lengend Snippet: Figure 6. Effect of Z. seed oil on MAPK signaling in the lung tissue of OVA‑induced asthmatic mice. (A) Western blot analysis of phosphorylated and unphosphorylated ERK1/2, p38 MAPK, JNK, Ras and β‑actin expression in vehicle‑ , OVA‑ and OVA + Z. seed oil‑treated mice. Quantitative analysis of the expression levels of (B) p‑ERK1/2, (C) p‑p38 MAPK, (D) p‑JNK and (E) Ras. Protein expression levels were normalized to β‑actin. Data are expressed as the mean ± standard deviation (n=3). *P<0.05 vs. the OVA‑treated asthmatic mice. OVA, ovalbumin; Z. seed oil, Zanthoxylum bungeanum seed oil; d, days; p‑ERK, phosphorylated‑extracellular signal regulated kinase; JNK, c‑jun N‑terminal kinase.

Article Snippet: Membranes were incubated with primary antibodies, as follows rabbit polyclonal anti-Ras (1:1,000; Cell Signaling Technology, Inc., Danvers, MA, USA; cat. no. 3965), rabbit polyclonal anti-ERK1/2 (1:1,000; Santa Cruz Biotechnology, Inc.; cat. no. sc-94), rabbit polyclonal anti-phosphorylated (p)-ERK1/2 (1:600; Santa Cruz Biotechnology, Inc.; sc-7383), rabbit polyclonal anti-p38 MAPK (1:800; Santa Cruz Biotechnology, Inc.; sc-7149), rabbit polyclonal anti-p-p38 MAPK (1:1,000; Cell Signaling Technology, Inc.; cat. no. 9211), rabbit polyclonal anti-JNK (1:800; Santa Cruz Biotechnology, Inc.; sc-572), rabbit polyclonal anti-nuclear factor-κB-p65 (NF-κB-p65; 1:1,000; Santa Cruz Biotechnology, Inc.; cat. no. sc-372), rabbit polyclonal anti-c-fos (1:1,000; Santa Cruz Biotechnology, Inc.; cat. no. sc-52), rabbit polyclonal anti-c-JUN (1:600; Santa Cruz Biotechnology, Inc.; cat. no. sc-1694), rabbit polyclonal β-actin (1:600; Wuhan Boster Biological Technology, Ltd., Wuhan, China; cat. no. MK1656), mouse monoclonal anti-p-JNK (1:600; Cell Signaling Technology, Inc.; cat. no. sc-6254), mouse monoclonal anti-p‐activating transcription factor-2 (ATF-2; 1:600; Santa Cruz Biotechnology, Inc.; cat. no. sc-8398), rabbit monoclonal anti-p-NF-κB‐p65 (1:1,000; Cell Signaling Technology, Inc.; cat. no. 3033), rabbit polyclonal intercellular adhesion molecule 1 (ICAM-1; 1:600; BIOSS; cat. no. bS-0608R), rabbit polyclonal anti-tumor necrosis factor-α (TNF-α; 1:600; BIOSS; cat. no. bS-00789), rabbit polyclonal anti-CD14 (1:600; Wuhan Boster Biological Technology, Ltd.; cat. no. BA0719-2), rabbit polyclonal anti-TNF-receptor (TNF-R; 1:600; BIOSS; cat. no. bS-2941R), rabbit polyclonal anti-toll-like receptor (TLR)2 (1:600; BIOSS; cat. no. bS-1019R) and rabbit polyclonal anti-TLR4 (1:600; BIOSS; cat. no. bS-1021R).

Techniques: Western Blot, Expressing, Standard Deviation

Journal: Molecular medicine reports

Article Title: Inhibitory effect of Zanthoxylum bungeanum seed oil on ovalbumin‑induced lung inflammation in a murine model of asthma.

doi: 10.3892/mmr.2016.5050

Figure Lengend Snippet: Figure 7. Effect of Z. seed oil on ERK1/2 and p38 MAPK nuclear levels in the lung tissue of OVA‑induced asthmatic mice. Western blot analysis and quan tification of (A and B) ERK1/2 and (C and D) p38 MAPK expression and phosphorylation levels in nuclear extracts from vehicle‑ , OVA‑ and OVA + Z. seed oil‑treated mice. Protein expression levels were normalized to β‑actin. qPCR analysis of (E) ERK1/2 and (F) p38 MAPK mRNA expression levels. Data are expressed as the mean ± standard deviation (n=3). *P<0.05 vs. the OVA‑treated asthmatic mice. OVA, ovalbumin; Z. seed oil, Zanthoxylum bungeanum seed oil; d, days; p‑ERK, phosphorylated‑extracellular signal regulated kinase.

Article Snippet: Membranes were incubated with primary antibodies, as follows rabbit polyclonal anti-Ras (1:1,000; Cell Signaling Technology, Inc., Danvers, MA, USA; cat. no. 3965), rabbit polyclonal anti-ERK1/2 (1:1,000; Santa Cruz Biotechnology, Inc.; cat. no. sc-94), rabbit polyclonal anti-phosphorylated (p)-ERK1/2 (1:600; Santa Cruz Biotechnology, Inc.; sc-7383), rabbit polyclonal anti-p38 MAPK (1:800; Santa Cruz Biotechnology, Inc.; sc-7149), rabbit polyclonal anti-p-p38 MAPK (1:1,000; Cell Signaling Technology, Inc.; cat. no. 9211), rabbit polyclonal anti-JNK (1:800; Santa Cruz Biotechnology, Inc.; sc-572), rabbit polyclonal anti-nuclear factor-κB-p65 (NF-κB-p65; 1:1,000; Santa Cruz Biotechnology, Inc.; cat. no. sc-372), rabbit polyclonal anti-c-fos (1:1,000; Santa Cruz Biotechnology, Inc.; cat. no. sc-52), rabbit polyclonal anti-c-JUN (1:600; Santa Cruz Biotechnology, Inc.; cat. no. sc-1694), rabbit polyclonal β-actin (1:600; Wuhan Boster Biological Technology, Ltd., Wuhan, China; cat. no. MK1656), mouse monoclonal anti-p-JNK (1:600; Cell Signaling Technology, Inc.; cat. no. sc-6254), mouse monoclonal anti-p‐activating transcription factor-2 (ATF-2; 1:600; Santa Cruz Biotechnology, Inc.; cat. no. sc-8398), rabbit monoclonal anti-p-NF-κB‐p65 (1:1,000; Cell Signaling Technology, Inc.; cat. no. 3033), rabbit polyclonal intercellular adhesion molecule 1 (ICAM-1; 1:600; BIOSS; cat. no. bS-0608R), rabbit polyclonal anti-tumor necrosis factor-α (TNF-α; 1:600; BIOSS; cat. no. bS-00789), rabbit polyclonal anti-CD14 (1:600; Wuhan Boster Biological Technology, Ltd.; cat. no. BA0719-2), rabbit polyclonal anti-TNF-receptor (TNF-R; 1:600; BIOSS; cat. no. bS-2941R), rabbit polyclonal anti-toll-like receptor (TLR)2 (1:600; BIOSS; cat. no. bS-1019R) and rabbit polyclonal anti-TLR4 (1:600; BIOSS; cat. no. bS-1021R).

Techniques: Western Blot, Expressing, Phospho-proteomics, Standard Deviation

Journal: Oncology reports

Article Title: Fenretinide inhibits the proliferation and migration of human liver cancer HepG2 cells by downregulating the activation of myosin light chain kinase through the p38‑MAPK signaling pathway.

doi: 10.3892/or.2018.6436

Figure Lengend Snippet: Figure 3. Effect of 4‑HPR on the migration ability and expression of p38‑MAPK in HepG2 cells. HepG2 cells were treated with 4‑HPR or ATRA at different concentrations for 48 h. (A) The treatment suppressed the migration of HepG2 cells. (a) Migration image. (b) Analysis of the migration rate. Calculation method: Relative migration ratio=(start distance‑end distance)/start distance. All values are presented as mean ± SD. n=6. 24 h: #P<0.05 compared with the cell group; 48 h: *P<0.05 compared with the cell group. (B) (a) Western blot analyses revealed that the phosphorylation of p38 was significantly increased. Lane 1, cell control; lane 2, DMSO; lane 3, 5 µM ATRA; lane 4, 10 µM ATRA; lane 5, 5 µM 4‑HPR; and lane 6, 10 µM 4‑HPR. (b) Analysis of contrast gray value. All values are presented as mean ± SD. n=3. *P<0.05 compared with the DMSO group; #P<0.05 compared with the ATRA group. 4‑HPR, fenretinide; ATRA, all‑trans retinoic acid.

Article Snippet: Primary antibodies: Rabbit antihuman monoclonal MLCK (cat. no. ab92721) and E-cadherin (cat. no. ab40772), mouse anti-human monoclonal F-actin (cat. no. ab205) were obtained from Abcam (Cambridge, MA, USA); rabbit anti‐human monoclonal phosphpho-p38 MAPK (cat. no. 4511), mouse anti-human monoclonal phospho-MLC (cat. no. 3675) were purchased from Cell Signaling Technology (Danvers, MA, USA); rabbit anti-human polyclonal MLC (cat. no. 15354-1-AP), mouse anti-human monoclonal GAPDH (cat. no. 60004-1-Ig) were obtained from Proteintech Group (Wuhan, Hubei, China); rabbit anti-human polyclonal p38 MAPK (cat. no. sc-7149) and mouse antihuman monoclonal β-actin (cat. no. sc-47778) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Migration, Expressing, Western Blot, Phospho-proteomics, Control

Journal: Oncology reports

Article Title: Fenretinide inhibits the proliferation and migration of human liver cancer HepG2 cells by downregulating the activation of myosin light chain kinase through the p38‑MAPK signaling pathway.

doi: 10.3892/or.2018.6436

Figure Lengend Snippet: Figure 4. Effect of 4‑HPR and SB203580 on the migration of HepG2 cells. The cells were treated with 4‑HPR or ATRA (10 µM) alone, or in combination with SB203580 (25 µM), for 48 h. 4‑HPR or ATRA joining with SB203580 significantly accelerated the migration of HepG2 cells. Calculating method as Fig. 3. All values are presented as mean ± SD. n=6. 24 h: #P<0.05 compared with 4‑HPR group; 48 h: *P<0.05 compared with 4‑HPR group and ΔP<0.05 compared with ATRA group. 4‑HPR, fenretinide; ATRA, all‑trans retinoic acid; SB, p38‑MAPK inhibitor SB203580.

Article Snippet: Primary antibodies: Rabbit antihuman monoclonal MLCK (cat. no. ab92721) and E-cadherin (cat. no. ab40772), mouse anti-human monoclonal F-actin (cat. no. ab205) were obtained from Abcam (Cambridge, MA, USA); rabbit anti‐human monoclonal phosphpho-p38 MAPK (cat. no. 4511), mouse anti-human monoclonal phospho-MLC (cat. no. 3675) were purchased from Cell Signaling Technology (Danvers, MA, USA); rabbit anti-human polyclonal MLC (cat. no. 15354-1-AP), mouse anti-human monoclonal GAPDH (cat. no. 60004-1-Ig) were obtained from Proteintech Group (Wuhan, Hubei, China); rabbit anti-human polyclonal p38 MAPK (cat. no. sc-7149) and mouse antihuman monoclonal β-actin (cat. no. sc-47778) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Migration

Journal: Oncology reports

Article Title: Fenretinide inhibits the proliferation and migration of human liver cancer HepG2 cells by downregulating the activation of myosin light chain kinase through the p38‑MAPK signaling pathway.

doi: 10.3892/or.2018.6436

Figure Lengend Snippet: Figure 7. Effect of 4‑HPR and ML‑7 on the expression of MLCK, E‑cadherin, F‑actin and phosphoryled MLC and p38. (A) ML‑7 was used to treat HepG2 cells. The phosphorylation of MLC was evidently decreased. In addition, the protein expression of MLC was decreased by ML‑7 combined with 4‑HPR or ATRA. All values are presented as mean ± SD. n=3, *P<0.05, ◊P<0.05 compared with DMSO group. (B) Phosphorylated (p)‑p38 was increased by ML‑7. All values are presented as mean ± SD. n=3, *P<0.05 compared with DMSO group, #P<0.05 compared with ATRA group and ΔP<0.05 compared with 4‑HPR group. (C) The protein expression of E‑cadherin was increased and F‑actin was decreased by ML‑7. All values are presented as mean ± SD. n=3, *P<0.05, #P<0.05 compared with DMSO group. For A‑C: lane 1, DMSO; lane 2, ML‑7; lane 3, ATRA; lane 4, ATRA+ML‑7; lane 5, 4‑HPR; lane 6, 4‑HPR+ML‑7. 4‑HPR, fenretinide; ATRA, all‑trans retinoic acid; ML‑7, a specific inhibitor of MLCK.

Article Snippet: Primary antibodies: Rabbit antihuman monoclonal MLCK (cat. no. ab92721) and E-cadherin (cat. no. ab40772), mouse anti-human monoclonal F-actin (cat. no. ab205) were obtained from Abcam (Cambridge, MA, USA); rabbit anti‐human monoclonal phosphpho-p38 MAPK (cat. no. 4511), mouse anti-human monoclonal phospho-MLC (cat. no. 3675) were purchased from Cell Signaling Technology (Danvers, MA, USA); rabbit anti-human polyclonal MLC (cat. no. 15354-1-AP), mouse anti-human monoclonal GAPDH (cat. no. 60004-1-Ig) were obtained from Proteintech Group (Wuhan, Hubei, China); rabbit anti-human polyclonal p38 MAPK (cat. no. sc-7149) and mouse antihuman monoclonal β-actin (cat. no. sc-47778) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Expressing, Phospho-proteomics

Journal: PLoS ONE

Article Title: A Mechanism of Male Germ Cell Apoptosis Induced by Bisphenol-A and Nonylphenol Involving ADAM17 and p38 MAPK Activation

doi: 10.1371/journal.pone.0113793

Figure Lengend Snippet: A) Seminiferous tubule cells from rats injected with 50 mg/kg of BPA or NP were isolated as described in the section and incubated with and antibody against the active form of ADAM17. The graph shows that the percentage of cells labeled with the antibody increases in rats treated with BPA or NP, but is reduced when rats were treated in the presence of GW280264X, a pharmacological inhibitor of ADAM17 (GW), or PD169316, a pharmacological inhibitor of p38 MAPK (PD). * p<0.05, n = 3.

Article Snippet: Rabbit polyclonal antibody against ADAM17 (ab39163), which reacts with an epitope located in the activation site (cysteine switch and furin cleavage site) of ADAM17 – , and p38 (phospho Y182+T 180) were purchased from Abcam (Cambridge, MA, USA), Rabbit polyclonal antibody against PARP-1/2 (sc-7150), clusterin-α (sc-8354), total-p38 MAPK (sc-7149) and mouse monoclonal SCP-3 (sc-74569) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Injection, Isolation, Incubation, Labeling

Journal: PLoS ONE

Article Title: A Mechanism of Male Germ Cell Apoptosis Induced by Bisphenol-A and Nonylphenol Involving ADAM17 and p38 MAPK Activation

doi: 10.1371/journal.pone.0113793

Figure Lengend Snippet: A) Time course of p38 MAPK phosphorylation in 21-day-old rats treated with 50 mg/kg of BPA. B) Time course of p38 MAPK phosphorylation in 21-day-old rats treated with 50 mg/kg of NP. C) Intra-testicular application of 5 µM PD169316 reduces the phosphorylation of p38 MAPK 1 or 2 h after in vivo treatment with BPA or NP. D) The pharmacological inhibitor of p38 MAPK (PD169316) prevents the increase of TUNEL-positive cells in testes of 21-day-old rats treated with 50 mg/kg of BPA or NP. * p<0.05, n = 3.

Article Snippet: Rabbit polyclonal antibody against ADAM17 (ab39163), which reacts with an epitope located in the activation site (cysteine switch and furin cleavage site) of ADAM17 – , and p38 (phospho Y182+T 180) were purchased from Abcam (Cambridge, MA, USA), Rabbit polyclonal antibody against PARP-1/2 (sc-7150), clusterin-α (sc-8354), total-p38 MAPK (sc-7149) and mouse monoclonal SCP-3 (sc-74569) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Phospho-proteomics, In Vivo, TUNEL Assay

Journal: Cell death & disease

Article Title: C1q-TNF-related protein-3 attenuates pressure overload-induced cardiac hypertrophy by suppressing the p38/CREB pathway and p38-induced ER stress.

doi: 10.1038/s41419-019-1749-0

Figure Lengend Snippet: Fig. 4 CTRP3 regulates activation of the p38 MAPK/CREB pathway in TAC mouse. a, b Representative western blot (top) and quantification (bottom) of the p38-CREB signaling pathway activity in the hearts of mice with different genotypes (WT, Ctrp3-KO, and LV-CTRP3) 4 weeks after sham treatment or TAC surgery (n = 5–6 mice per group). c, d Representative immunofluorescence images of murine heart sections stained with p-p38 (red) and DAPI (blue) (left), and the percentage of p-p38–positive nuclei (right) in the hearts of mice with different genotypes (WT, Ctrp3-KO, and LV- CTRP3) 4 weeks after sham treatment or TAC surgery (n = 5–6 mice per group). The data were analyzed by one-way ANOVA. *p < 0.05, **p < 0.01 vs. SHAM, #p < 0.05 vs. TAC. In the bar graphs, the data are presented as the mean ± SEM

Article Snippet: The primary antibodies used were anti-p-CREB (Ser133; 9198 s), anti-CREB (9197 s), anti-p-CaMKII (12716), antip-eIF2α (3398), anti-eIF2α (5324), anti-ATF4 (11815), anti-CHOP (2895), anti-p-AMPKα (2537), anti-AMPKα (5832), anti-p-AKT (4060), anti-AKT (9272), anti-Bcl-2 (3498), and anti-Bax (5023) antibodies, purchased from Cell Signaling Technology (Beverly, MA, USA); anti-ANP (ab209232), anti-CaN A (ab3673), anti-CaMK□ (ab52476), anti-GRP78 (ab21685), anti-p-IRE1 (ab48187), anti-IRE1 (ab37073), anti-XBP1 (ab37152), anti-ATF6 (ab203119), and anti-CTRP3 (ab36870) antibodies, purchased from Abcam (Cambridge, MA, USA); anti-p-p38 (sc-166182), anti-p38 (sc-7149), and anti-MYH7 (sc53089) antibodies, purchased from Santa Cruz Biotechnology (Santa Cruz, CA, China); and anti-GAPDH (AT0002) antibody purchased from cmcTAG (Milwaukee, WI, USA).

Techniques: Activation Assay, Western Blot, Activity Assay, Staining

Journal: Cell death & disease

Article Title: C1q-TNF-related protein-3 attenuates pressure overload-induced cardiac hypertrophy by suppressing the p38/CREB pathway and p38-induced ER stress.

doi: 10.1038/s41419-019-1749-0

Figure Lengend Snippet: Fig. 5 P38 MAPK activation mediates CTRP3 deficiency-aggravated cardiac hypertrophy and fibrosis. a Ejection fraction (EF)% after 4 weeks of TAC in the indicated groups (n = 5–6 mice per group). b The HW/BW ratio in animals from indicated groups after 4 weeks of TAC (n = 5–6 mice per group). c Representative images of the heart sections stained with HE, WGA, and Masson stain (n = 5 mice per group). d The mean cross-sectional area of cardiomyocytes from the indicated groups (n ≥100 cells per group). e The LV collagen volume in different groups (n ≥40 fields per group). f Real-time PCR analysis of the expression of genes encoding the hypertrophic markers β-MHC, ANP, and BNP, and the fibrotic markers TGF-β1, collagen-I, and collagen-III in each group (n = 5 mice per group). g Representative western blot (top) and quantification (bottom) of the CREB activity in the hearts of mice from indicated groups (n = 5 mice per group). The data were analyzed by one-way ANOVA. *p < 0.05 between the two indicated groups; ns, not significant. In the bar graphs, the data are presented as the mean ± SEM

Article Snippet: The primary antibodies used were anti-p-CREB (Ser133; 9198 s), anti-CREB (9197 s), anti-p-CaMKII (12716), antip-eIF2α (3398), anti-eIF2α (5324), anti-ATF4 (11815), anti-CHOP (2895), anti-p-AMPKα (2537), anti-AMPKα (5832), anti-p-AKT (4060), anti-AKT (9272), anti-Bcl-2 (3498), and anti-Bax (5023) antibodies, purchased from Cell Signaling Technology (Beverly, MA, USA); anti-ANP (ab209232), anti-CaN A (ab3673), anti-CaMK□ (ab52476), anti-GRP78 (ab21685), anti-p-IRE1 (ab48187), anti-IRE1 (ab37073), anti-XBP1 (ab37152), anti-ATF6 (ab203119), and anti-CTRP3 (ab36870) antibodies, purchased from Abcam (Cambridge, MA, USA); anti-p-p38 (sc-166182), anti-p38 (sc-7149), and anti-MYH7 (sc53089) antibodies, purchased from Santa Cruz Biotechnology (Santa Cruz, CA, China); and anti-GAPDH (AT0002) antibody purchased from cmcTAG (Milwaukee, WI, USA).

Techniques: Activation Assay, Staining, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Activity Assay

Journal: Cell death & disease

Article Title: C1q-TNF-related protein-3 attenuates pressure overload-induced cardiac hypertrophy by suppressing the p38/CREB pathway and p38-induced ER stress.

doi: 10.1038/s41419-019-1749-0

Figure Lengend Snippet: Fig. 8 CTRP3 exerted its protective role on pathological cardiac hypertrophy through inhibiting p38/CREB/ER stress pathway. a Representative western blot (left) and quantification (right) of the ER stress signaling pathway activity in NRCMs from the indicated groups (n = 4 samples per group). b Representative western blot (left) and quantification (right) of the p38-CREB and ER stress signaling pathway activity in NRCMs from the indicated groups (n = 4 samples per group). c Representative western blot (left) and quantification (right) of ER stress and p38-CREB signaling pathway activity, protein expression levels of ANP and β-MHC in NRCMs from the indicated groups (n = 4 samples per group). *p < 0.05, **p < 0.01 between indicated groups. In the bar graphs, the data are presented as the mean ± SEM. d Proposed protective mechanism of the role of CTRP3 in pathological cardiac hypertrophy. CTRP3 inhibits the activation of the p38/CREB signaling pathway and ER stress pathway in cardiomyocytes under pressure overload, thereby alleviating pathological cardiac hypertrophy

Article Snippet: The primary antibodies used were anti-p-CREB (Ser133; 9198 s), anti-CREB (9197 s), anti-p-CaMKII (12716), antip-eIF2α (3398), anti-eIF2α (5324), anti-ATF4 (11815), anti-CHOP (2895), anti-p-AMPKα (2537), anti-AMPKα (5832), anti-p-AKT (4060), anti-AKT (9272), anti-Bcl-2 (3498), and anti-Bax (5023) antibodies, purchased from Cell Signaling Technology (Beverly, MA, USA); anti-ANP (ab209232), anti-CaN A (ab3673), anti-CaMK□ (ab52476), anti-GRP78 (ab21685), anti-p-IRE1 (ab48187), anti-IRE1 (ab37073), anti-XBP1 (ab37152), anti-ATF6 (ab203119), and anti-CTRP3 (ab36870) antibodies, purchased from Abcam (Cambridge, MA, USA); anti-p-p38 (sc-166182), anti-p38 (sc-7149), and anti-MYH7 (sc53089) antibodies, purchased from Santa Cruz Biotechnology (Santa Cruz, CA, China); and anti-GAPDH (AT0002) antibody purchased from cmcTAG (Milwaukee, WI, USA).

Techniques: Western Blot, Activity Assay, Expressing, Activation Assay