Journal: bioRxiv

Article Title: Mechanical Ventilation induced DNA Damage and P21 in an Acute Aging Model of Lung Injury

doi: 10.1101/2022.03.08.483505

Figure Lengend Snippet: Immunofluorescence staining of lung section for both young and old mice. Staining for KRT8 (red), a marker of transiently differentiated AT2, and SFTPC (green), an AT2 marker. DAPI (blue) was used as a counterstain. The staining reveals an increase in KRT8 positive cells with age and mechanical ventilation and a decrease in SFTPC in all groups compared to young non-ventilated. NV: non-ventilated and HP: high pressure.

Article Snippet: Primary antibodies: mouse anti-gamma H2AX (Novus Biologicals, NB100-74435), rabbit anti-Ki67/MKi67 (Novus Biologicals, NB500-170SS), rabbit anti-SP-C (Bioss, bs-10067R), rat anti-KRT8 (CreativeBiolabs, CBDH1469).

Techniques: Immunofluorescence, Staining, Marker

Journal: Journal of integrative neuroscience

Article Title: Kv1.3 Blockade Alleviates White Matter Injury through Reshaping M1/M2 Phenotypes via the NF-κB Signaling Pathway after Intracerebral Hemorrhage.

doi: 10.31083/j.jin2206171

Figure Lengend Snippet: Fig. 9. Application of PAP-1 inhibiting NF-κB activation after ICH. (A) Bar charts showing NF-κB P65 mRNA expression in each group on day 3 after ICH. n = 3 each group, ∗∗p < 0.01 vs. Sham group; #p < 0.05 vs. ICH group. (B) Bar charts showing NF-κB

Article Snippet: Then the sample was incubated separately with the following primary antibodies overnight at 4 °C: Rabbit anti-Kv1.3 (1:1000, No.14079-1-AP, Proteintech, Philadelphia, PA, USA); Rabbit anti-MBP (1:1000, No.BA0094, Boster, Wuhan, Hubei, China); Rabbit antiCD16 (1:1000, No.16559-1-AP, Proteintech, Philadelphia, PA, USA); Rabbit anti-CD206 (1:1000, No.18704-1-AP Proteintech, Philadelphia, PA, USA); Rabbit anti-iNOS (1:1000, No.22226-1-AP, Proteintech, Philadelphia, PA, USA); Rabbit anti-Arg-1 (1:1000, No.16001-1-AP, Proteintech, Philadelphia, PA, USA); Mouse anti-TNF-α (1:1000, No.sc-52746, Santa Cruz, Dallas, TX, USA); Mouse anti-IL-1β (1:1000, No.sc-32294, Santa Cruz, Dallas, TX, USA); Rabbit anti-IL-4 (1:1000, No.66142-1Ig, Proteintech, Philadelphia, PA, USA); Rabbit anti-IL10 (1:1000, No.60269-1-Ig, Proteintech, Philadelphia, PA, USA); Mouse anti-NF-κB p65 (1:200, No.sc-8008, Santa Cruz, Dallas, TX, USA); Mouse anti-NF-κB p50 (1:200, No.sc-8414, Santa Cruz, Dallas, TX, USA); Mouse antip-NF-κB p65 (1:200, No.sc-136548, Santa Cruz, Dallas, TX, USA); Mouse anti-p-NF-κB p50 (1:200, No.sc271908, Santa Cruz, Dallas, TX, USA); Mouse anti-βTubulin (1:5000, No.T200608, Zen-bio, Chengdu, Sichuan, China); Mouse anti-GAPDH (1:5000, No.250133, Zen-bio, Chengdu, Sichuan, China); Mouse anti-β-Actin (1:5000, No.700068, Zen-bio, Chengdu, Sichuan, China).

Techniques: Activation Assay, Expressing

Journal: PloS one

Article Title: Pre-clinical characterization of Dacomitinib (PF-00299804), an irreversible pan-ErbB inhibitor, combined with ionizing radiation for head and neck squamous cell carcinoma.

doi: 10.1371/journal.pone.0098557

Figure Lengend Snippet: Figure 1. EGFR is over-expressed in SCCHN cell lines. qRT-PCR was performed to measure ErbB transcript levels in the three SCCHN cell lines, normalized to that of the normal oral epithelial (NOE) cells. Expression fold change was determined by the 22DDCt method. ***p, 0.001; student’s t test. doi:10.1371/journal.pone.0098557.g001

Article Snippet: Phospho-EGFR immunohistochemistry was performed using microwave antigen retrieval in combination with the Level-2 Ultra Streptavidin System and antip-EGFR antibody (1:100 dilution; #3777, Cell Signalling), as previously described [28].

Techniques: Quantitative RT-PCR, Expressing

Journal: PloS one

Article Title: Pre-clinical characterization of Dacomitinib (PF-00299804), an irreversible pan-ErbB inhibitor, combined with ionizing radiation for head and neck squamous cell carcinoma.

doi: 10.1371/journal.pone.0098557

Figure Lengend Snippet: Figure 2. Dacomitinib demonstrated in vitro efficacy in EGFR over-expressing SCCHN cell lines. (A to D) Growth inhibition of three SCCHN and the NOE cell lines, treated with Dacomitinib, with or without IR. (B) FaDu; (C) UT-SCC-8; (D) UT-SCC-42a; and (E) NOE cells were treated with Dacomitinib (0, 10, 25, 50, 100, or 250 nM) alone, or in combination with IR (0, 2, or 4 Gy). MTS assays were conducted 72 hours post-treatment. The graphs represent data from 3 independent experiments, with the mean 6 SEM reported. *p,0.05, **p,0.01, ***p,0.001; one-way ANOVA. No statistically significant difference between curves for Figure 2A-D; two-way ANOVA. Combination of Dacomitinib plus RT did not result in a synergistic interaction for any of the dosing regimens in all three SCCHN cell lines (Chou-Talalay Method). doi:10.1371/journal.pone.0098557.g002

Article Snippet: Phospho-EGFR immunohistochemistry was performed using microwave antigen retrieval in combination with the Level-2 Ultra Streptavidin System and antip-EGFR antibody (1:100 dilution; #3777, Cell Signalling), as previously described [28].

Techniques: In Vitro, Expressing, Inhibition

Journal: PloS one

Article Title: Pre-clinical characterization of Dacomitinib (PF-00299804), an irreversible pan-ErbB inhibitor, combined with ionizing radiation for head and neck squamous cell carcinoma.

doi: 10.1371/journal.pone.0098557

Figure Lengend Snippet: Figure 4. Treatment of SCCHN cells with Dacomitinib inhibited EGFR signalling. FaDu, UT-SCC-8, and UT-SCC-42a cells were treated with Dacomitinib (D; 0, 10, 50, 100, 250, or 500 nM) in serum-free media for 24 hours. Thirty minutes prior to lysis, cells were stimulated with EGF (20 ng/ mL). Experiments were performed three independent times, with similar results; representative blots are shown. doi:10.1371/journal.pone.0098557.g004

Article Snippet: Phospho-EGFR immunohistochemistry was performed using microwave antigen retrieval in combination with the Level-2 Ultra Streptavidin System and antip-EGFR antibody (1:100 dilution; #3777, Cell Signalling), as previously described [28].

Techniques: Lysis

Journal: PloS one

Article Title: Pre-clinical characterization of Dacomitinib (PF-00299804), an irreversible pan-ErbB inhibitor, combined with ionizing radiation for head and neck squamous cell carcinoma.

doi: 10.1371/journal.pone.0098557

Figure Lengend Snippet: Figure 6. Dacomitinib delayed FaDu tumor growth, alone and in combination with IR. (A) FaDu-bearing mice were randomized to: (i) DMSO control; (ii) Dacomitinib (5 mg/kg/d) delivered orally on days 1 to 5; (iii) local tumor irradiation (IR; 2 Gy/treatment) delivered on days 2 and 5; or (iv) Dacomitinib plus IR. Mean tumor leg diameter (TLD) from three independent experiments (three mice per treatment group, per experiment) is reported 6 SEM. ***p,0.001, negative control vs. Dacomitinib only, or RT only vs. Dacomitinib plus RT; Student’s t test. (B) Mean mouse weight 6 SEM from experiment (A). (C) Ki-67 staining was performed on treated FaDu tumors: DMSO, radiation (IR), Dacomitinib (D), or Dacomitinib plus radiation (D+IR), as from (A). Tumors were extracted from mice 24 hours after the final treatment. Ki-67 positive cells stain dark brown. The right-hand panel represents the proportion of Ki-67-positive cells for each treatment group. Ki-67 scoring was conducted by counting the number of positive tumor cells in 3 representative sections for each tumor. Mean 6 SEM is reported. ***p,0.001; Student’s t test. (D) p-EGFR staining was performed on paraffin sections, as in (C). Positive Staining for p-EGFR is brown. doi:10.1371/journal.pone.0098557.g006

Article Snippet: Phospho-EGFR immunohistochemistry was performed using microwave antigen retrieval in combination with the Level-2 Ultra Streptavidin System and antip-EGFR antibody (1:100 dilution; #3777, Cell Signalling), as previously described [28].

Techniques: Control, Irradiation, Negative Control, Staining