Journal: STAR Protocols

Article Title: Protocol to establish cancer-associated fibroblasts from surgically resected tissues and generate senescent fibroblasts

doi: 10.1016/j.xpro.2021.100553

Figure Lengend Snippet:

Article Snippet: Rabbit pSTQ , Cell Signaling Technology , Cat# 2851.

Techniques: Recombinant, Labeling, Software

Journal: Cancer Management and Research

Article Title: KLF7 Promotes Gastric Carcinogenesis Through Regulation of ANTXR1

doi: 10.2147/CMAR.S308071

Figure Lengend Snippet: Downregulation of KLF7 suppresses xenografted tumour growth. ( A ) MKN45 cell lines expressing shCtrl or shKLF7 were injected into athymic nude mice (n=5) and analysed for tumour formation. Representative images of xenografted tumours formed with shCtrl or shKLF7 MKN45 cells (2×106 cells/mouse). ( B ) The average tumour volumes at the indicated time points. ( C ) Quantification of the weight of tumours formed with shCtrl or shKLF7 MKN45 cells (2×106 cells/mouse). *** p <0.001, **p<0.01. ( D ) Western blot was performed to determine the expression levels of KLF7 and ANTXR1 in xenografted tumours after KLF7 knockdown.

Article Snippet: Next, the proteins were transferred to nitrocellulose membranes and blocked with 5% non-fat milk at room temperature for 1 h. The membranes were incubated with KLF7 (1:2000; Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA), ANTXR1 (1:2000; Proteintech), and GAPDH (1:5000; Proteintech) at 4°C overnight, followed by incubation with a secondary antibody (Thermo Fisher Scientific) at room temperature for 60 min.

Techniques: Expressing, Injection, Western Blot, Knockdown

Journal: Cancer Management and Research

Article Title: KLF7 Promotes Gastric Carcinogenesis Through Regulation of ANTXR1

doi: 10.2147/CMAR.S308071

Figure Lengend Snippet: KLF7 regulates cancer proliferation and migration through ANTXR1. ( A ) Correlation between KLF7 and ANTXR1 mRNA expression in stomach adenocarcinoma tissues (n=375) from TCGA database was analysed using the StarBase v2.0 database. ( B ) Correlation between the probability of survival for patients with gastric cancer and ANTXR1 expression. ( C and D ) qRT-PCR and Western blotting analysis of ANTXR1 expression in cells transfected with siCtrl and siKLF7. ANTXR1 was downregulated in cells transfected with siKLF7. For qRT-PCR, the expression levels were normalized to those of β-actin. For Western blotting, GAPDH was used as a loading control. ( E and F ) ANTXR1 was upregulated upon the overexpression of KLF7. qRT-PCR and Western blotting analysis of ANTXR1 expression in Ctrl and KLF7-overexpressing cells. For qRT-PCR, the expression levels were normalized to those of β-actin. For Western blotting, GAPDH was used as a loading control. ( G ) Western blotting analysis of AGS cells transfected with control, KLF7, or KLF7+siANTXR1. GAPDH was used as a loading control. ( H ) CCK-8 cell viability assay of AGS cells transfected with control, KLF7 or KLF7+siANTXR1. ( I and J ) Transwell invasion analysis of indicated AGS cells. ( K and L ) Colony formation assay of indicated AGS cells. *** p <0.001, **p<0.01.

Article Snippet: Next, the proteins were transferred to nitrocellulose membranes and blocked with 5% non-fat milk at room temperature for 1 h. The membranes were incubated with KLF7 (1:2000; Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA), ANTXR1 (1:2000; Proteintech), and GAPDH (1:5000; Proteintech) at 4°C overnight, followed by incubation with a secondary antibody (Thermo Fisher Scientific) at room temperature for 60 min.

Techniques: Migration, Expressing, Quantitative RT-PCR, Western Blot, Transfection, Control, Over Expression, CCK-8 Assay, Viability Assay, Colony Assay

Journal: Biochimica et biophysica acta. Molecular basis of disease

Article Title: Hepatocyte-like cells differentiated from methylmalonic aciduria cblB type induced pluripotent stem cells: A platform for the evaluation of pharmacochaperoning.

doi: 10.1016/j.bbadis.2022.166433

Figure Lengend Snippet: Fig. 3. Disease biomarkers in differentiated control and MMAB HLCs. A. Immunofluorescent detection of ATR (red) and the mitochondrial protein ATP5a (green) in C1 and MMAB1 HLCs (40× objective; scale bar 50 μm). The image shows the co-localization of both proteins in the mitochondria. B. Western blot immunodetection of ATR in C1, C2, MMAB1 and MMAB2 HLCs. GAPDH was used as a loading control. C. Quantification of methylmalonic acid in spent culture medium from healthy controls (mean of C1 and C2 methylmalonic acid levels) and diseased (MMAB1 and MMAB2) HLCs, with respect to the protein concentration in each corresponding sample. Values are fold changes with respect to controls. Differences were analyzed using the Student t-test (*** p < 0.001). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The antibodies used were those against MMAB (ProteinTech, Rosemont, IL, USA; 1:200), ATP5a (Abcam, Cambridge, UK; 1:200), OCT4 (Santa Cruz Biotechnology, Dallas, TX, USA, 1:60), SOX17 (Santa Cruz Biotechnology; 1:200), FOXA2 (Santa Cruz Biotechnology, 1:200), HNF4α (Santa Cruz Biotechnology, 1:100), AFP (Santa Cruz Biotechnology, 1:50), ALB (Cedarlane, Burlington, Ontario, Canada; 1:50) and E-Cadherin (Cell Signaling, Danvers, MA, USA; 1:200).

Techniques: Control, Western Blot, Immunodetection, Protein Concentration