Journal: BMC Cancer

Article Title: DOCK1 regulates the malignant biological behavior of endometrial cancer through c-Raf/ERK pathway

doi: 10.1186/s12885-024-12030-1

Figure Lengend Snippet: DOCK1 modulated the expression of related proteins and signaling pathway of endometrial cancer cells. ( A ) The expression of E-cadherin, MMP9, Ezrin and Bcl-2 protein was analyzed after DOCK1 knockout or overexpression. ( B ) The effect of DOCK1 knockout or overexpression on c-Raf/ERK signaling pathway related protein

Article Snippet: The primary antibodies (1:1000) were shown as follows: rabbit anti-Bcl2 antibody (Boster, China), rabbit anti-DOCK1 antibody (CST, USA), rabbit anti-E-cadherin antibody (Proteintech, USA), rabbit anti-Ezrin antibody (Proteintech, USA), mouse anti-GAPDH antibody (CST, USA), rabbit anti-MMP9 antibody (CST, USA), rabbit anti-tubulin antibody (ABclonal, China), rabbit anti-phospho-C-Raf (S338) antibody (CST, USA), rabbit anti-C-Raf antibody (CST, USA), rabbit anti-phospho-ERK1/2 (Thr202/Tyr204) antibody (CST, USA), rabbit anti-ERK1/2 antibody (CST, USA).

Techniques: Expressing, Knock-Out, Over Expression

Journal: BMC Cancer

Article Title: DOCK1 regulates the malignant biological behavior of endometrial cancer through c-Raf/ERK pathway

doi: 10.1186/s12885-024-12030-1

Figure Lengend Snippet: The inhibition of DOCK1 attenuated the growth of xenograft tumors in vivo. ( A-B ) The images of nude mice and subcutaneous tumors at 4 weeks after injecting DOCK1-KO Ishikawa cells. ( C-D ) The growth and weights of tumors were analyzed in DOCK1-KO group and control group. ( E ) The expression of E-cadherin, MMP9, Ezrin and Bcl-2 protein on xenograft tumors by immunohistochemical assay. * P < 0.05

Article Snippet: The primary antibodies (1:1000) were shown as follows: rabbit anti-Bcl2 antibody (Boster, China), rabbit anti-DOCK1 antibody (CST, USA), rabbit anti-E-cadherin antibody (Proteintech, USA), rabbit anti-Ezrin antibody (Proteintech, USA), mouse anti-GAPDH antibody (CST, USA), rabbit anti-MMP9 antibody (CST, USA), rabbit anti-tubulin antibody (ABclonal, China), rabbit anti-phospho-C-Raf (S338) antibody (CST, USA), rabbit anti-C-Raf antibody (CST, USA), rabbit anti-phospho-ERK1/2 (Thr202/Tyr204) antibody (CST, USA), rabbit anti-ERK1/2 antibody (CST, USA).

Techniques: Inhibition, In Vivo, Control, Expressing, Immunohistochemical staining

Journal: Cancer biology & therapy

Article Title: The involvement of MALAT1-ALKBH5 signaling axis into proliferation and metastasis of human papillomavirus-positive cervical cancer.

doi: 10.1080/15384047.2023.2249174

Figure Lengend Snippet: Figure 6. Effect of the MALAT1-ALKBH5 axis on MMP2 and MMP9 expression. (a and b) HeLa and CaSki cells were, respectively, transfected with NC and two siRNAs against MALAT1 or ALKBH5. The expression levels of MMP2 and MMP9 was determined by RT-qPCR. (c) The cell lysates were subjected to western blot analysis of MMP2 and MMP9 after knockdown of MALAT1 in HeLa and CaSki cells using GAPDH as loading control. (d) Both HeLa and CaSki cells were transfected with NC and siRNAs against MALAT1 with co-transfection of the ALKBH5 expression vector (pcDNA3.1-ALKBH5), with empty vector (pcDNA3.1) serving as the control. Cell RNA was isolated and subsequently reverse transcribed into cDNA, which were subjected to RT-qPCR for determination of MMP2 and MMP9 expression using specific primers. NC, negative control.

Article Snippet: The antibody information was as follows: Primary antibody: anti-ALKBH5 antibody (1:1000, Cell Signaling Technology, Inc.), anti-MMP2 antibody (1:1000, Elabscience Inc.), anti-MMP9 antibody (1:1000, BOSTER Biological Technology Co. Ltd.), and anti-GAPDH antibody (1:5000, Beyotime Institute of Biotechnology); and secondary antibody: HRP conjugated anti-rabbit or -mouse secondary antibody (1:5000, MultiSciences Biotech Co., Ltd.).

Techniques: Expressing, Transfection, Quantitative RT-PCR, Western Blot, Knockdown, Control, Cotransfection, Plasmid Preparation, Isolation, Reverse Transcription, Negative Control

Journal: Redox biology

Article Title: Coronin 1a-mediated F-actin disassembly controls effector function in murine neutrophils.

doi: 10.1016/j.redox.2025.103618

Figure Lengend Snippet: Fig. 3. Enhanced F-actin levels in Coro1a-deficient neutrophils are associated with impaired granule release (A-E) Granule exocytosis (degranulation) by WT and Coro1a−/−(KO) neutrophils. (A, B) Release of myeloperoxidase (MPO) from primary granules into culture supernatants upon stimulation of neutrophils with (A) serum-opsonized whole glucan particles (WGP) or (B) serum-opsonized C. albicans hyphae (SO–CaH). (C) Upregulation of CD63 (Lamp3) cell surface expression upon stimulation with WGP. (D, E) Release of (D) neutrophil gelatinase-associated lipocalin (NGAL) from secondary granules or (E) matrix metalloproteinase-9 (MMP-9) from tertiary granules into culture supernatants upon stimulation of neutrophils with TNFα. (F) Total amount of MPO, NGAL, and MMP-9 in WT and Coro1a−/−(KO) neutrophils expressed in % of WT (n = 5). Unpaired Student’s t-test. (G–I) PMA (100 ng/ml)-induced release of MMP-9 by WT and Coro1a−/−(KO) neutrophils pretreated for 60 min with (G) 16 μM jasplakinolide (Jas; actin-stabilizing agent), (H) 1 μM latrunculin B (Lat; actin-depolymerizing agent); or (I) 30 μM benproperine phosphate (Benp; Arp2/3-complex inhibitor). Data are mean ± SEM (A, D-E: n = 3; B, G-H: n = 2; C: n = 7). (A-E, G-H) 2-way ANOVA. ns > 0.05, * <0.05, ** <0.01, *** <0.001, **** <0.0001. Data are representative for at least 3 independent experiments.

Article Snippet: MMP-9 was detected with biotinylated anti-MMP9 polyclonal Ab (R&D Systems, BAF909) followed by staining with Streptavidin-AF488 (Thermo Fisher Scientific, S11223).

Techniques: Expressing

Journal: Redox biology

Article Title: Coronin 1a-mediated F-actin disassembly controls effector function in murine neutrophils.

doi: 10.1016/j.redox.2025.103618

Figure Lengend Snippet: Fig. 4. Local F-actin destruction is required for neutrophil granule exocytosis (A) Transmission electron microscopy (TEM) images of WT and Coro1a−/−(KO) neutrophils either unstimulated or challenged with TNFα (20 ng/ml; 30 min). Micrograph labels: primary (1◦G), secondary – quaternary (2◦G – 4◦G) granules, mitochondria (Mt). TNFα treated condition: non-primary granules are labeled with yellow asterisks. Scale bar, 2 μm; each micrograph is 4.5 μm × 4.5 μm, x22000 magnification. Images shown are representative of at least n = 50 neutrophils being analyzed from each condition. (B) Airyscan-enhanced confocal micrographs of WT and Coro1a−/−(KO) neutrophils in basal state (left panel) or stimulated with 20 ng/ml of TNFα for 30 min (right panel). Neutrophils were fixed permeabilized and stained for F-actin (phalloidin) and MMP-9 together with DAPI (nucleus). Scale bar: 3 μm. Images shown are representative of neutrophils observed in 3 independent microscopy experiments. Each experiment consisted of analysis of 8–12 fields for every condition, with each field containing ca. 10–30 cells.

Article Snippet: MMP-9 was detected with biotinylated anti-MMP9 polyclonal Ab (R&D Systems, BAF909) followed by staining with Streptavidin-AF488 (Thermo Fisher Scientific, S11223).

Techniques: Transmission Assay, Electron Microscopy, Labeling, Staining, Microscopy

Journal:

Article Title:

doi:

Figure Lengend Snippet: Gingival inflammation was present in both chimeras after gingival application of LPS (A-F). The degree of gingival inflammation was confirmed by the number of inflammatory infiltrates (A) and identification of IL1β (B), TNFα (C), IL6 (D), MCP1 (E), and MMP9 (F) producing cells. Inflammatory infiltrates were quantitatively measured using sections stained with H&E (n = 4 per treatment group) (A). Number of cells expressing cytokines (B, C, D), MCP1(E), and MMP9 (F) was assessed by immunofluorescence microscopy (n = 4 per treatment group). Inflammatory infiltrates were present in both GFPWT:WT and GFPWT:KO mice treated with LPS (A). Insets show inflammatory cell infiltrates. Arrows point to inflammatory cells. The number of inflammatory cells was significantly higher in LPS treated animals compared to PBS treated animals in both chimeric groups (p< 0.01) (A). There was no statistical difference in the amount of infiltrates between these chimeras with LPS application (A). There was also no statistical difference in the number of cells stained with IL1β, TNFα, IL6, MCP1 and MMP9 (B-F) between GFPWT:WT and GFPWT:KO mice with LPS application. Arrows indicate GFP+ donor cells expressing IL1β (B), TNFa (C), IL6 (D), MCP1 (E), and MMP9 (F) respectively. Inflammatory cell infiltrates were counted in 5 random fields visualized by light microscopy at 400X magnification. Photos of H&E sections were taken at 200X magnification. Insets are at 400X (A). Cells positive for cytokines, MCP1 and MMP9 were counted in 5 random fields and visualized by immunofluorescence microscopy at 400X magnification. Photos of immunofluorescence microscopy were taken at 400X (B-F). In all cases, the examiner was blinded to the animal group to which the tissue section belonged. x-axis: chimeric group. y-axis: number of cells/random field. Data presented are mean ± SD. (G) Alveolar bone loss was assessed by microtomography: Microtomography of maxilla from GFPWT:WT chimeras following 18 weeks of LPS or PBS treatment. The distances between the cemento-enamel junction (CEJ) and the alveolar crest are shown in red. The distance between the CEJ and the alveolar bone shown in a GFPWT:WT chimera without LPS application is occupied by junctional epithelium and connective tissue attached to the root and therefore does not reflect actual bone loss (G-a). Comparison of bone loss among chimeric groups (G-b):The bracket on the right bottom indicates the mean distance where epithelia and connective tissue attach. The bracket on the top indicates the mean bone loss. Bone loss was statistically greater in GFPWT:WT mice with LPS compared to PBS treatment. There is no statistically significant difference in bone loss between PBS and LPS treated groups in GFPWT:KO animals (G-b). x-axis: chimeric group, y-axis: distance (mm) measured from CEJ to the crest of alveolar bone on the mid palatal portion of the second maxillary molars. n = 8–12 per chimeric group (4–6 per treatment group). Data presented are mean ± SD. *p<0.05.

Article Snippet: The following primary antibodies were used at 1:100 dilution to detect cells producing IL1β, TNFα, IL6, MCP1, and MMP9 in the gingiva: rabbit polyclonal anti-IL1β (ab9722) (Abcam, Cambridge, MA), anti-TNFα (ab66579) (Abcam), anti-MCP1 sc-28879) (Santa Cruz Biotechnology, Dallas, TX), anti-MMP9 antibody ab124513) (Abcam).

Techniques: Staining, Expressing, Immunofluorescence, Microscopy, Light Microscopy