primary antibodies against flag, β-actin, p-ikkα/β, p-erk, p-p38  (Cell Signaling Technology Inc)

Journal: Journal of Advanced Research

Article Title: NAT10 promotes osteoclastogenesis in inflammatory bone loss by catalyzing Fos mRNA ac4C modification and upregulating MAPK signaling pathway

doi: 10.1016/j.jare.2024.07.031

Figure Lengend Snippet: Inhibition of NAT10 represses RANKL-activated MAPK signaling pathway. (A) Western blotting and quantification analysis of p-NFκB p65 and NFκB p65 in DMSO or Remodelin (20 μM)-pretreated RAW264.7 cells after RANKL stimulation at the designated times (n = 4). (B) Western blotting and quantification analysis of p-ERK1/2, ERK1/2, p-JNK, JNK, p-p38 and p38 in DMSO or Remodelin (20 μM)-pretreated RAW264.7 cells after RANKL stimulation at the designated times (n = 4). (C) Western blotting and quantification analysis of p-ERK1/2, ERK1/2, p-JNK, and JNK in DMSO or Remodelin (20 μM)-pretreated BMDMs after RANKL stimulation at the designated times (n = 4). (D) Western blotting and quantification analysis of p-ERK1/2, ERK1/2, p-JNK, and JNK in shNC and sh Nat10 RAW264.7 cells after RANKL stimulation at the designated times (n = 4). (E) The SRE-Luc and AP-1-Luc activities in DMSO or Remodelin (20 μM)-pretreated RAW264.7 cells after RANKL stimulation for 1 h (n = 4). Data were shown in the form of mean ± SD, two-way ANOVA was performed with Bonferroni’s multiple comparisons test, and Mann-Whitney U test was performed for comparisons between two groups. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001.

Article Snippet: Primary antibodies against β-actin, p-NFκB p65, NFκB p65, p-ERK1/2, ERK1/2, p-JNK, JNK, p-p38, and p38 were from Cell Signaling Technology, NAT10 was from Abcam (Cambridge, UK), ACP5, c-Jun, and c-Fos were from Proteintech (Wuhan, China), and NFATc1 was from Santa Cruz (CA, USA).

Techniques: Inhibition, Western Blot, MANN-WHITNEY

Journal: ACS applied materials & interfaces

Article Title: Tailored Mesoporous Silica Nanoparticles and the Chick Chorioallantoic Membrane: A Promising Strategy and Model for Efficient Blood-Brain Barrier Crossing.

doi: 10.1021/acsami.5c05429

Figure Lengend Snippet: Figure 4. In vitro Dox loading, release, and cytotoxicity in U87 Cells treated with Dox-loaded MSNs and transport of Dox across the BBB in the chick CAM. (a) Loading efficiency and loading amount of Dox-loaded MSNs. (b) Cumulative release profile of Dox from MSMs at pH 7.4 and 5.5. (c) Cytotoxicity analysis of U87 cells treated with Dox alone, MSNs (at equivalent Dox-loaded MSN concentration), and Dox-loaded MSNs (at equivalent Dox alone concentration) across various concentrations (1−20 μg/mL). Cell viability was measured using a CCK-8 assay 24 h after treatment. (d) Western blot analysis showing protein expression levels of Phospho-p38 and HMGB1. α-Tubulin was used as a loading control. (e) IVIS imaging of Dox distribution. (f) Radiant efficiency of Dox in chick embryonic brains. N = 10. (g) Quantitative analysis of Dox distribution in brain sections using TissueQuest software. Quantitative analysis of tissue staining and identification of nucleus-positive events were performed using TissueFAXS and TissueQuest software platforms, respectively (red: Dox; blue: DAPI-stained nuclei). (h) Confocal microscopy images of brain sections showing Dox distribution (red) and nuclei (blue, DAPI staining). Scale bar: 20 μm. (i) Drug permeability assay. Quantification of Dox content in brain homogenates using fluorescence spectrophotometry. Data are presented as mean ± SD. Statistical significance was analyzed using Student’s t test (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001).

Article Snippet: The PVDF membrane was blocked in PBS containing 0.1% Tween 20 (PBS-T) with 5% (w/v) BSA for 1 h and then washed three times with PBS-T. Primary antibodies against p-p38 (sc-17852-R, Santa Cruz Biotechnology, USA), HMGB1 (ab227168, abcam, UK), and αtubulin (sc-5286, Santa Cruz Biotechnology, USA) were incubated with the PVDF membrane at 4 °C overnight with gentle rocking.

Techniques: In Vitro, Concentration Assay, CCK-8 Assay, Western Blot, Expressing, Control, Imaging, Software, Staining, Confocal Microscopy, Permeability, Fluorescence, Spectrophotometry

Journal: Biomedicines

Article Title: Patient-Derived Cancer-Associated Fibroblasts Support the Colonization of Tumor Cells in Head and Neck Squamous Cell Carcinoma

doi: 10.3390/biomedicines13020358

Figure Lengend Snippet: Western blot analysis of p-p38 and vimentin in SCC-25 and SCC25-E cells with and without irradiation. The original Western blot image of ( A ) p-p38 and ( B ) vimentin compared to the loading control GAPDH in control SCC-25 cells (1), in the mixed culture at passages 1 (2) and 3 (3), in irradiated SCC-25 cells (4), and in irradiated SCC25-E cells (p3) (5). Densitometry of ( C ) p-p38 and ( D ) vimentin normalized to the GAPDH loading control. The normalized optical density of the control SCC-25 cells was considered 100%, and the columns show the relative normalized optical density.

Article Snippet: IHC for phosphorylated p38 MAPK was performed using the rabbit monoclonal primary antibody against p-p38 MAPK (Cat. Nr. 4511S; 1:400), purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Western Blot, Irradiation, Control

Journal: Biomedicines

Article Title: Patient-Derived Cancer-Associated Fibroblasts Support the Colonization of Tumor Cells in Head and Neck Squamous Cell Carcinoma

doi: 10.3390/biomedicines13020358

Figure Lengend Snippet: ( A ) Identification of the brown positive staining reaction of p-p38 in the nuclei of tumor cells in the original HNSCC cancer cell nest. ( B ) Cytoplasmic p-p38 antigen detection of single cells in the stromal compartment of the sectioned HNSCC tissue. Brown reactions without connections to cell nuclei are artifacts. The bottom left box of each image shows an overview of the whole scanned tissue section. The red box in each image shows the location of the presented area in the original tissue section.

Article Snippet: IHC for phosphorylated p38 MAPK was performed using the rabbit monoclonal primary antibody against p-p38 MAPK (Cat. Nr. 4511S; 1:400), purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Staining