Journal: Drug Design, Development and Therapy

Article Title: Inhibition of Pre-B Cell Colony Enhancing Factor Reduces Lung Injury in Rats Receiving Cardiopulmonary Bypass

doi: 10.2147/DDDT.S281554

Figure Lengend Snippet: Adenovirus-encoding sh-PBEF reduced the increased phosphorylation of ERK1/2, Akt, and p38MAPK in rat lung tissue. ( A ) Representative blots; ( B ) Phosphorylation of ERK1/2; ( C ) Phosphorylation of AKT; ( D ) Phosphorylation of p38MAPK vs control; Original blots were shown in Supplemental figure 1.*P<0.05; vs LPS, # P<0.05; vs CPB+Blank, @ P<0.05.

Article Snippet: The membrane was incubated with the following antibodies overnight at 4°C: rabbit anti-PBEF (1:2000, 11776-1-AP, Proteintech); rabbit anti-ERK1/2 (1:1000, 16443-1-AP, Proteintech); rabbit anti-p-ERK1/2 (1:500, bs-3016R, Bioss); rabbit anti-p38MAPK (1:1000, bs-0637R, Bioss); rabbit anti-p-p38MAPK (1:1000, bs-5477R, Bioss); rabbit anti-AKT (1:500, ab8805, Abcam); rabbit anti-p-AKT (1:1000, bs-2720R, Bioss); rabbit anti-AQP1 (1:1000, bs-1506R, Bioss); rabbit anti-AQP5 (1:1000, AF5169, Affinity); rabbit anti-ENaC (1:1000, bs-2957R, Bioss); rabbit anti-surfactant protein D (SP) (1:1000, ab220422, Abcam); and mouse monoclonal anti-GAPDH (1:2000, TA-08, ZSBio).

Techniques:

Journal: ACS Omega

Article Title: Injectable Thermosensitive Hydrogel Containing Resveratrol-Laden Nanoparticles Promotes Hypertrophic Scar Repair in Rat Tail

doi: 10.1021/acsomega.5c01469

Figure Lengend Snippet: (A) Representative immunofluorescence-staining images of phosphorylated p38-MAPK expression in scar tissue at week 0, week 2, and week 6 post-treatments. Scale bar = 200 or 50 μm. (B) ROS activity in scar tissue at week 0, week 2, and week 6 post-treatments. (C) Quantitative evaluation of phosphorylated p38-MAPK expression in scar tissue. Notes: compared with normal skin (NS), ** P < 0.01, *** P < 0.001,**** P < 0.0001; compared with NC, △△ P < 0.01, △△△△ P < 0.0001; compared with MSN@Res, #### P < 0.0001.

Article Snippet: Subsequently, they were kept at 4 °C overnight in the presence of rabbit antiphosphorylated p38-MAPK (p-p38, bs-5476R, Bioss, China, 1:400), rabbit anti-LC3-II (LC3-II, 14600–1-AP, Aspen, China, 1:500), rabbit anti-cleaved caspase-3 (Cleaved Caspase 3, 25128–1-AP, Aspen, China, 1:150), rodent-derived p53 antibody (p53, 60283–2-IG, Aspen, China, 1:400), and a rat-originated HIF-1α antibody (HIF-1α, SC-13515, Santa Cruz, 1:100).

Techniques: Immunofluorescence, Staining, Expressing, Activity Assay

Journal: BMC Oral Health

Article Title: Artesunate-loaded thermosensitive chitosan hydrogel promotes osteogenesis of maxillary tooth extraction through regulating T lymphocytes in type 2 diabetic rats

doi: 10.1186/s12903-024-04127-7

Figure Lengend Snippet: ART-loaded TCH suppressed MAPK signaling pathway in the maxilla of T2DM tooth extraction. A Network pharmacology analysis revealed the potential immunoregulatory mechanism of ART in T2DM. The top ten enriched pathways were illustrated. B Molecular docking of ART to MAPK and ERK1 proteins. C, D The protein levels of p38-MAPK, p-p38-MAPK, ERK1, and p-ERK1, were analyzed by WB and IHC ( n = 3), which can be downregulated by ART-loaded TCH treating. The uncropped blots are displayed in Additional file . E Relative mRNA expression of MAPK ( D ) and ERK1 ( E ) was analyzed by PCR ( n = 3). The 150 mg/ml ART-loaded TCH group significantly decreased the mRNA expression of MAPK and ERK1 than the T2DM group ( p < 0.01). Data are shown as mean ± standard deviation. * p < 0.05 and ** p < 0.01. T2DM, type 2 diabetes mellitus group; TCH, thermosensitive chitosan hydrogel group; Low, 50 mg/ml ART-loaded TCH group; Medium, 100 mg/ml ART-loaded TCH group; High; 150 mg/ml ART-loaded TCH group

Article Snippet: The membranes were then incubated overnight with primary antibodies including rabbit anti-β-actin (1:20000, Bioss), mouse anti-GATA-3 (1:2000, Proteintech), mouse anti-IL-4 (1:1000, Proteintech), rabbit anti-T-BET (1:500, Proteintech), rabbit anti-phospho-p38 MAPK (1:1000, Bioss), rabbit anti-p38 MAPK (1:1000, Bioss), mouse anti-ERK1 (1:1000, Bioss), mouse anti-phospho-ERK1 (1:1000, Bioss).

Techniques: Extraction, Expressing, Standard Deviation

Journal: Oncotarget

Article Title: Klotho inhibits EGF-induced cell migration in Caki-1 cells through inactivation of EGFR and p38 MAPK signaling pathways

doi: 10.18632/oncotarget.25481

Figure Lengend Snippet: (A) Cells were pretreated with EGF (100ng/ml) and harvested after the indicated times. Lysates were collected, and p38 MAPK phosphorylation levels were analyzed by Western blotting using the antiphospho-p38 antibody. The same membrane was striped and re-probed with a different antibody for measuring total-p38 MAPK protein levels. (B) Caki-1 cells were preincubated with 400pM of the secreted form of the Klotho (KL) protein at the indicated times followed by EGF (100ng/ml) stimulation for additional 5 min. Cell lysates were subjected to Western blot analysis as described in (A). Plots indicate means ± S.E.M of three independent experiments.

Article Snippet: Rabbit anti-human polyclonal active p38 MAPK antibody and SB203580 were purchased from Promega (Madison, WI).

Techniques: Phospho-proteomics, Western Blot, Membrane

Journal: Oncotarget

Article Title: Klotho inhibits EGF-induced cell migration in Caki-1 cells through inactivation of EGFR and p38 MAPK signaling pathways

doi: 10.18632/oncotarget.25481

Figure Lengend Snippet: (A) Classical in vitro wound healing assay was performed using near-confluent serum-starved Caki-1 cells grown on collagen type 1. Cells were either pretreated with 400pM Klotho (KL) or 1μM p38 MAPK-specific inhibitor SB203580 as positive control, for 60 min followed by EGF (100ng/ml) treatment. Cell culture images shown here were taken at 0 and 24 h. (B) Plots of quantification of the resultant cell motility values were computed by gap surface area measurements for four selected microscopic fields in each assay condition. The degree of migration is expressed as % wound closure compared with zero time point. The results represent means ± S.E.M of three independent experiments. (C) Wound healing assays performed under 3D settings. Images are microphotographs of Caki-1 cells showing hole-closure of “tissue openings” generated with magnetic pattering as described in the materials and method. Cells were pretreated the same way with either Klotho (KL) or SB203580 and stimulated with EGF as described for the classical wound healing assay. (D) Plots of rate closure of holes for Caki-1 cells as a function of EGF exposure in the presence or absence of KL or SB203580.

Article Snippet: Rabbit anti-human polyclonal active p38 MAPK antibody and SB203580 were purchased from Promega (Madison, WI).

Techniques: In Vitro, Wound Healing Assay, Positive Control, Cell Culture, Migration, Generated