Journal: Nature Communications

Article Title: Novel role of the synaptic scaffold protein Dlgap4 in ventricular surface integrity and neuronal migration during cortical development

doi: 10.1038/s41467-022-30443-z

Figure Lengend Snippet: a Representative RPE1 cell images showing reduced lamellipodia morphology and increased filopodia elongation (white arrows) after WT or mutant DLGAP4 transfections. Yellow arrows: F-actin. b Quantifications of cell percentages with lamellipodia morphology, c filopodia elongation (µm) and d organized stress fibers. Quantification data ( b – d ) represents the individual values, mean ± SEM (~100 cells per condition, n = 3 independent experiments). One-way ANOVA with post hoc Tukey’s test was performed. b F 2, 6 = 88.8, p < 0.0001; ** p = 0.0029, ***p = 0.0007, ****p < 0.0001. ( c ) F 2, 6 = 7.21, p = 0.025; Ctl vs WT DLGAP4 : *p = 0.036 and Ctl vs MUT DLGAP4 : *p = 0.041. d F 2, 6 = 1.25, p = 0.35. e IP performed from N e uro2A cells co-transfected with Flag-cortactin and GFP- DLGAP4 (WT or MUT). f Quantification data represent the relativized individual values, means ± SEM ( n = 2 WT; n = 3 MUT), ** p = 0.0018, two-sided unpaired t-test with Welch’s correction. g Flag-cortactin was found in GFP-DLGAP4 IPs. h Western blot assays from RPE1 cell extracts transfected with WT or MUT GFP-DLGAP4. Endogenous levels of DLG1, p38, FLNA, Raptor and Rictor were detected and normalized to GAPDH. i Table summarizing results. Dashed arrows: trend. One-way ANOVA with post hoc Tukey and at least three independent experiments were performed. DLGAP4 ( n = 5): F 2, 12 = 6.51, p = 0.012; control vs WT, p = 0.011, control vs MUT , p = 0.079. DLG1 ( n = 3): F 2, 6 = 0.7, p = 0.53. p38 ( n = 3): F 2, 6 = 147.3, p < 0.0001; control vs WT and control vs MUT, p < 0.0001. FLNA ( n = 4): F 2, 9 = 5.25, p = 0.031; control vs WT, p = 0.26 ; control vs MUT, p = 0.025. Raptor ( n = 3): F 2, 6 = 25.18, p = 0.0012; control vs WT, p = 0.0028, control vs MUT, p = 0.0016. Rictor ( n = 3): F 2, 6 = 6.49, p = 0.032; control vs WT, p = 0.98, control vs MUT, p = 0.042. j – l HA-Raptor and WT or MUT Flag-DLGAP4 co-IP in Neuro2A cells ( n = 2).

Article Snippet: Endogenous protein levels were detected by the following primary antibodies, applied O/N at 4 °C: mouse anti-GFP (G6539, Sigma-Aldrich, 1:1000), anti-DLG1 (clone S64-15, Abnova, 1:1000), rabbit anti-DLGAP4 (Ab73285, Abcam, 1:500), anti-Raptor (ab5454, Abcam, 1:2000), anti-Rictor (A300-459A, Bethyl, 1:2000), anti-FlnA (A301-135A, Bethyl, 1:2000), anti-p38 (SPC-172, StressMarq Bioscience, 1:1000).

Techniques: Mutagenesis, Transfection, Western Blot, Co-Immunoprecipitation Assay

Journal: International Journal of Clinical and Experimental Pathology

Article Title: Hawthorne leaf flavonoids prevent oxidative stress injury of renal tissues in rats with diabetic kidney disease by regulating the p38 MAPK signaling pathway

doi:

Figure Lengend Snippet: Immunochemical analysis of the (A) p38MAPK (B) and p-p38MAPK expression in the kidney of different groups (200×). Quantitative analysis of (C) p38MAPK and (D) p-p38MAPK expression in the kidney of different groups. The expression of p38MPAK and p-p38MAPK was significantly higher in the DKD group than in the HLF and IRB groups.

Article Snippet: Rabbit anti-rat p38MAPK and p-p38MAPK polyclonal antibodies were purchased from Boster (Beijing, China).

Techniques: Expressing

Journal: Redox Report : Communications in Free Radical Research

Article Title: The novel thioredoxin reductase inhibitor butaselen suppresses lung cancer by inducing oxidative stress

doi: 10.1080/13510002.2025.2588086

Figure Lengend Snippet: BS affects NF-κB, p38/JNK, and PI3K-Akt signaling pathways in lung cancer. (a–d) A549 and H1299 cells were treated with BS (0, 5, 10 μM) for 24 h, followed by western blot. Error bars are means ± std. * P < 0.05, ** P < 0.01, *** P < 0.001, compared with the control group by One-way ANOVA (n = 3).

Article Snippet: The primary antibodies used for immunoblotting analysis are as follows: TrxR1 (Proteintech, 11117-1-AP), Trx1 (Proteintech, 14999-1-AP), HBP1 (Proteintech, 11746-1-AP), DNMT1 (Proteintech, 24206-1-AP), Bcl-2 (Proteintech, 12789-1-AP), Bax (Proteintech, 50599-2-Ig), β-actin (Bioss, bs-0061R), Flag (Sigma-Aldrich, F1804), HA (Covance, MMS-101P), p53 (Santa, sc-126), p21 (MBL, K0081-3), p27 (MBL, K0082), γ-H2AX (CST, 9718), NF-κB (Abcam, ab32536), p-NF-κB(CST, 3033), p38 (Santa, sc-7972), p-p38 (Santa, sc-101759), JNK (MCE, HY- P80728 ), p-JNK (Immunoway, YP0157), Akt (Santa, sc-5298), phospho-Akt (CST, 4060).

Techniques: Protein-Protein interactions, Western Blot, Control

Journal: Redox Report : Communications in Free Radical Research

Article Title: The novel thioredoxin reductase inhibitor butaselen suppresses lung cancer by inducing oxidative stress

doi: 10.1080/13510002.2025.2588086

Figure Lengend Snippet: Schematic model of lung cancer inhibition by BS. The TrxR/Trx inhibitor butaselen (BS) can inhibit lung cancer by inducing ROS-dependent apoptosis. The inactivation of the NF-κB and PI3K-Akt signaling pathways, along with the activation of the p38/JNK signaling pathway, contributes to the anti-cancer effects of BS on lung cancer. Although p53 itself is not activated by BS, the HBP1/DNMT1/p21/γ-H2AX/Bcl-2/Bax signaling pathway is activated by BS and contributes to its tumor-inhibitory role. Further mechanistic studies revealed HBP1 as a novel target of the Trx system. The Trx system inversely associates with HBP1 in lung cancer and regulates HBP1 expression at the post-translational level. Under normal conditions, TrxR1 catalyzes the reduction of Trx1 by utilizing NADPH. In its reduced form, Trx1 interacts with HBP1, promoting the ubiquitination of HBP1, which leads to its proteasomal degradation and maintains a low level of HBP1 within cancer cells. Treatment with butaselen inhibits the activity of TrxR1 in lung cancer cells, resulting in the oxidation of Trx1 and the subsequent excessive generation of ROS. HBP1 is activated after being released by the oxidized Trx1 and escaping proteasomal degradation. The activated HBP1 inhibits the expression of DNMT1 and elevates Bax. The decreased DNMT1 further results in the demethylation of the whole genome as well as the promoters of p21 and HOXA9. Ultimately, the upregulation of p21 and γ-H2AX, along with the downregulation of DNMT1 and Bcl-2/Bax, contributes to the apoptosis of lung cancer cells induced by BS. Taken together, the TrxR/Trx inhibitor butaselen inhibits lung cancer by promoting ROS-induced apoptosis through the NF-κB, PI3K-Akt, p38/JNK, and HBP1/DNMT1 signaling pathways.

Article Snippet: The primary antibodies used for immunoblotting analysis are as follows: TrxR1 (Proteintech, 11117-1-AP), Trx1 (Proteintech, 14999-1-AP), HBP1 (Proteintech, 11746-1-AP), DNMT1 (Proteintech, 24206-1-AP), Bcl-2 (Proteintech, 12789-1-AP), Bax (Proteintech, 50599-2-Ig), β-actin (Bioss, bs-0061R), Flag (Sigma-Aldrich, F1804), HA (Covance, MMS-101P), p53 (Santa, sc-126), p21 (MBL, K0081-3), p27 (MBL, K0082), γ-H2AX (CST, 9718), NF-κB (Abcam, ab32536), p-NF-κB(CST, 3033), p38 (Santa, sc-7972), p-p38 (Santa, sc-101759), JNK (MCE, HY- P80728 ), p-JNK (Immunoway, YP0157), Akt (Santa, sc-5298), phospho-Akt (CST, 4060).

Techniques: Inhibition, Protein-Protein interactions, Activation Assay, Expressing, Ubiquitin Proteomics, Activity Assay

Journal: Molecular medicine reports

Article Title: Placental growth factor gene silencing mitigates the epithelial‑to‑mesenchymal transition via the p38 MAPK pathway in rats with hyperoxia‑induced lung injury.

doi: 10.3892/mmr.2019.10785

Figure Lengend Snippet: Figure 5. Expression of p38 and p‑p38 MAPK in hyperoxia‑exposed lung tissue. Neonatal rats were exposed to normoxic or hyperoxic conditions for 14 days; hyperoxia exposed rats were subsequently injected with shRNA‑NC or shRNA‑PLGF. (A) Western blot analysis and (B) quantification was performed to determine p‑p38/p38 protein expression levels in rat lung tissues; β‑actin was used as the endogenous control. Data are presented as the mean ± standard deviation. n=8. ***P<0.001. MAPK, mitogen‑activated protein kinase; NC, negative control; PLGF, placental growth factor; shRNA, short hairpin RNA.

Article Snippet: The animals were housed at a temperature of 25‐27 ̊C, with a humidity of 50-70% and a 12 h light/dark cycle with ad libitum access to food and water. xylene, absolute ethanol, eosin y and hydrogen peroxide were purchased from Wuhan uScn Business co., ltd. Hematoxylin, eosin and goat serum (cat. no. Sl038) were purchased from Beijing Solarbio Science& Technology co., ltd. PlGF mouse monoclonal antibody (cat. no. sc-518003) and e-cadherin mouse monoclonal antibody (cat. no. sc-71007) were purchased from Santa cruz Biotechnology, inc. anti-phosphorylated (p)-p38 rabbit polyclonal antibody (cat. no. bs-2210r) was purchased from (BioSS). anti-p38 rabbit monoclonal (cat. no. M00176), anti-β-actin goat polyclonal (cat. no. BM0627) and anti-ZeB2 rabbit polyclonal (cat. no. Pa1959) antibodies were purchased from Boster Biological Technology.

Techniques: Expressing, Injection, Western Blot, Control, Standard Deviation, Negative Control, shRNA

Journal: Frontiers in pharmacology

Article Title: Neuroprotective Effects and Mechanisms of Zhenlong Xingnao Capsule in In Vivo and In Vitro Models of Hypoxia.

doi: 10.3389/fphar.2019.01096

Figure Lengend Snippet: FIGURE 6 | Effects of ZXC on the mRNA levels of the Bcl-2/Bax ratio (A), caspase-3 (B), nuclear factor (NF)-кB (C), and p38 (D) in the prefrontal cortex of ischemia-reperfusion injury rats. The data are expressed as mean ± standard deviation (n = 3). I-30, ischemia for 30 min; I-90, ischemia for 90 min; I-90+R-30, ischemia for 90 min, then reperfusion for 30 min; I-90+R-180, ischemia for 90 min, then reperfusion for 180 min. *P < 0.05 vs. sham group; #P < 0.05 vs. model group; ##P < 0.01 vs. model group.

Article Snippet: The primary antibodies used for IHC were the MAPK14 (p38) antibody and CASP3 (P17) antibody; both were obtained from Boster Biological Technology Co., Ltd. (Wuhan, China).

Techniques: Standard Deviation

Journal: Frontiers in pharmacology

Article Title: Neuroprotective Effects and Mechanisms of Zhenlong Xingnao Capsule in In Vivo and In Vitro Models of Hypoxia.

doi: 10.3389/fphar.2019.01096

Figure Lengend Snippet: FIGURE 7 | Western blotting results for Sham, Model and ZXC groups (A). Effects of ZXC on the protein expressions Bcl-2 (B), Bax (C), Caspase-3 (D), NF-кB (E), and p38 (F) in brain tissue of ischemia-reperfusion injury rats induced by MCAO. The data are expressed as mean ± standard deviation (n = 4). I-30, ischemia for 30 min; I-90, ischemia for 90 min; I-90+R-30, ischemia for 90 min, then reperfusion for 30 min. *P < 0.05 vs. sham group; #P < 0.05 vs. model group.

Article Snippet: The primary antibodies used for IHC were the MAPK14 (p38) antibody and CASP3 (P17) antibody; both were obtained from Boster Biological Technology Co., Ltd. (Wuhan, China).

Techniques: Western Blot, Standard Deviation