Journal: Oncology Letters

Article Title: Silencing of GRHL2 induces epithelial‑to‑mesenchymal transition in lung cancer cell lines with different effects on proliferation and clonogenic growth

doi: 10.3892/ol.2023.13977

Figure Lengend Snippet: GRHL2 knockdown promotes the phosphorylation of AKT and ERK in HBEC4KT and lung cancer cell lines. Western blotting of GRHL2, p- or total AKT and p- or total ERK in HBEC4KT, H1975 and H2009 cells transfected with control or GRHL2 siRNA. Values below bands indicate the semi-quantitation of band intensities normalized to AKT for p-AKT (ser473), ERK for p-ERK or β-actin for GRHL2. GRHL2, Grainyhead-like 2; siRNA, small-interfering RNA; NC, negative control; p-, phosphorylated.

Article Snippet: The primary antibodies used were rabbit anti-GRHL2 (cat. no. HPA062839; Merck KGaA; 1:500), mouse anti-E-cadherin (cat. no. 610181; BD Biosciences; 1:1,000), mouse anti-vimentin (cat. no. HPA001762; Merck KGaA; 1:500), rabbit anti-AKT (pan; cat. no. 4691; Cell Signaling Technology, Inc.; 1:1,000), rabbit anti-phosphorylated (p-)-AKT (ser473; cat. no. 4060; Cell Signaling Technology, Inc.; 1:1,000), rabbit anti-p44/42MAPK (Erk1/2; cat. no. 9102; Cell Signaling Technology, Inc.; 1:1,000), rabbit anti-p-p44/42MAPK (Erk1/2; cat. no. 4370; Cell Signaling Technology, Inc.; 1:1,000) and mouse anti-β-actin (cat. no. A2228; Merck KGaA; 1:2,000).

Techniques: Western Blot, Transfection, Quantitation Assay, Small Interfering RNA, Negative Control

Journal: bioRxiv

Article Title: CCT and Cullin1 regulate the TORC1 pathway to promote dendritic arborization in health and disease

doi: 10.1101/2023.07.31.551324

Figure Lengend Snippet: ( A ) Heat map showing percent change in Raptor fluorescence of CCT5-IR or Raptor-IR knockdowns, as well as Raptor OE as compared to controls. ( B ) S6k fluorescence is significantly reduced in S6k-IR conditions as compared to WT. ( C ) P-Akt fluorescence is significantly reduced in Akt-IR conditions. ( D ) Cullin1 fluorescence is significantly reduced in Cullin1-IR conditions as compared to WT. ( E ) Heat map showing percent change in β -tubulin IIA for each genetic manipulation. Each experimental condition was compared to WT control and appropriate statistical comparisons were performed. In all panels * = p < 0.05, see for detailed statistics.

Article Snippet: Primary antibodies used include: chicken anti-GFP (1:1000 dilution, Aves Labs), rabbit anti-phosphorylated S6k (1:300 dilution, Cell Signaling Technology), rabbit anti-Raptor (1:200 dilution, Cell Signaling Technology), mouse anti-acetylated α-tubulin (1:100 dilution, Santa Cruz Biotechnology), mouse anti-Futsch (1:100 dilution, Developmental Studies Hybridoma Bank), rabbit anti-Huntingtin (1:200 dilution, Cell Signaling Technology), rabbit anti-HA (1:500 dilution, Cell Signaling Technology), rat anti-CCT1 (1:200 dilution, Origene), mouse anti-CCT5 (1:200 dilution, GeneTex), mouse anti-S6k (1:200 dilution, Proteintech), rabbit anti-phosphorylated Akt (1:200 dilution, Cell Signaling Technology), rabbit anti-Cullin1 (1:200 dilution, Invitrogen), mouse anti-β-tubulin IIA (1:500 dilution, Novus Biologicals).

Techniques: Fluorescence

Journal: International Journal of Oncology

Article Title: A disintegrin and metalloprotease 12 contributes to colorectal cancer metastasis by regulating epithelial-mesenchymal transition

doi: 10.3892/ijo.2023.5498

Figure Lengend Snippet: Effects of ADAM12 on oncogenic signaling pathways in the human colorectal cancer cells. Representative western blot images and semi-quantification graphs for the phosphorylation levels of PDK1, GSK-3β and AKT in AV-transfected or AS-transfected DLD1 or SW480 cells. Data are presented as the mean ± SD (n=3). * P<0.05. ADAM12, a disintegrin and metalloprotease 12; PDK1, phosphoinositide-dependent protein kinase 1; GSK-3β, glycogen synthase kinase-3β; EV, empty pcDNA6-myc vector; AV, ADAM12-pcDNA6-myc construct; SS, scrambled siRNA; AS, ADAM12 siRNA; siRNA, small interfering RNA; p-, phosphorylated; T-, total.

Article Snippet: Antibodies against E-cadherin (cat. no. #14472), Snail (cat. no. #3879), vimentin (cat. no. #5741), claudin-1 (cat. no. #4933), integrin α5 (cat. no. #4705), integrin β1 (cat. no. #9699), integrin β3 (cat. no. #13166), phosphorylated (p)-AKT (S473) (cat. no. #4060), p-phosphoinositide-dependent protein kinase 1 (PDK1) (S241) (cat. no. #3438), p-glycogen synthase kinase-3β (GSK-3β) (S9) (cat. no. #9323), total AKT (cat. no. #4691), total PDK1 (cat. no. #3062), total GSK-3β (cat. no. #9832) and Myc-tag (cat. no. #2278) were obtained from Cell Signaling Technology, Inc. Antibodies against β-tubulin (cat. no. sc-9104) and GAPDH (cat. no. sc-25778) were purchased from Santa Cruz Biotechnology, Inc.

Techniques: Western Blot, Transfection, Plasmid Preparation, Construct, Small Interfering RNA