3 5 full race kit  (TaKaRa)

Journal: bioRxiv

Article Title: Identification of genes supporting cold resistance of mammalian cells: lessons from a hibernator

doi: 10.1101/2023.12.27.573489

Figure Lengend Snippet: (A) Cold resistance of hamster HapT1 cells but not of human HT1080 cells. The proportion of dead cells was determined by LDH assay (One-way ANOVA with the Tukey’s multiple comparison test, p < 0.05). (B) Schematic illustration of hamster cDNA expression screening for genes that render the resistance to cold-rewarming culture to human HT1080 cells. (C) Venn diagram of the genes inserted in the genome of survived cells under cold-rewarming culture in four screening experiments. (D) The proportion of dead cells in HT1080 cells after 24hr of cold (4°C) culture when each indicated gene was overexpressed (N = 3 wells, One-way ANOVA with the Dunnett’s multiple comparison test, ***p < 0.001).

Article Snippet: The full-length cDNA expression library was constructed using the SMARTer RACE 5’/3’ Kit (Takara Bio, 634858) according to the manufacturer’s instructions for the In-Fusion SMARTer Directional cDNA Library Construction Kit (Takara Bio, 634933).

Techniques: Lactate Dehydrogenase Assay, Comparison, Expressing

Journal: Frontiers in Immunology

Article Title: CD4 and LAG-3 from sharks to humans: related molecules with motifs for opposing functions

doi: 10.3389/fimmu.2023.1267743

Figure Lengend Snippet: Alignment of the deduced amino acid sequences of CD4 and LAG-3 molecules in representative species. Predicted signal peptides are in Italic font and shaded in gray. Residue numberings under the alignment follow human CD4 or human LAG-3 mature proteins. Gray shading within the ectodomain (and not in Italic font) highlights β-strands as indicated for human CD4 and mouse LAG-3 in PDB accessions 1WIP and 8DGG, respectively, and names for β-strand regions are indicated with the letters A-to-G. The numbers between brackets refer to introns and to their phases at the indicated position (0) or in the preceding codon (1,2); intron positions that can only be speculated because genomic DNA sequence information is not available for that species are indicated with “(?)”. Cysteines are in purple and, based on Hopp and Woods, 1981 , red font is used for basic residues, blue for acidic residues, and of the other residues (green and orange) the more hydrophilic ones are in green. Highlighted in the cytoplasmic tail are the motifs Cx(C/H) in CD4 (yellow/orange), P(K/Q)P(K/R)(A/G)FY(H/K/R) in CD4-1, and FPAL(D/E) in LAG-3 (cyan). The species for which sequences are shown are Chgr: Chiloscyllium griseum (gray bambooshark); Gici: Ginglymostoma cirratum (nurse shark); Scca: Scyliorhinus canicula (small-spotted catshark); Scto: Scyliorhinus torazame (cloudy catshark); Heze: Heterodontus zebra (zebra bullhead shark); Pose: Polypterus senegalus (gray bichir); Erca: Erpetoichthys calabaricus (Reedfish); Acru: Acipenser ruthenus (sterlet sturgeon); Posp: Polyodon spathula (Mississippi paddlefish); Leoc: Lepisosteus oculatus (spotted gar); Dare: Danio rerio (zebrafish); Icpu: Ictalurus punctatus (channel catfish); Onmy: Oncorhynchus mykiss (rainbow trout); Taru: Takifugu rubripes (fugu); Lame: Latimeria menadoensis (Menado coelacanth); Lach: Latimeria chalumnae (West Indian Ocean Coelacanth); Pran: Protopterus annectens (West African lungfish); Xetr: Xenopus tropicalis (tropical clawed frog); Chmy: Chelonia mydas (green sea turtle); Gaga: Gallus gallus (chicken); Mumu: Mus musculus (mouse); Hosa: Homo sapiens (human). The Scyliorhinus torazame (cloudy catshark) CD4 intron positions were determined by comparison of the experimentally determined cDNA sequence with the rather short (in most cases containing only one exon) genomic sequences available as GenBank accessions BFAA01030515, BFAA01044711, BFAA01027384, BFAA01122682, BFAA01153775, BFAA01050327, BFAA01052865, and BFAA01028059; for LAG-3 in this species this was done similarly using GenBank accessions BFAA01031787, BFAA01335495, BFAA01051587, BFAA01228759, and BFAA01053433.

Article Snippet: For determining sequences, total RNA from gill tissue was reverse-transcribed into cDNA for 5’- and 3’-RACE PCR with SMARTer RACE 5’/3’ Kit (Takara Bio) for full-length cDNA cloning of catshark CD4 and LAG-3 genes.

Techniques: Residue, Sequencing, Comparison, Genomic Sequencing

Journal: Advances in clinical and experimental medicine : official organ Wroclaw Medical University

Article Title: LncRNA-LINC00472 suppresses the malignant progression of non-small cell lung cancer via modulation of the miRNA-1275/Homeobox A2 axis.

doi: 10.17219/acem/168431

Figure Lengend Snippet: Fig. 1. LINC00472 is downregulated in non-small cell lung cancer (NSCLC) and predominantly located in the cytoplasm. A. Volcano map of differential genes in NSCLC gene expression chip GSE44077. X-axis represents the log10 p-value, while Y-axis represents the logFC value; the red points represent significantly upregulated genes in the tumor, the green points represent markedly downregulated genes in the tumor, and the black points represent genes with no significant difference; B. LINC00472 level in normal (left) and tumor (right) groups in GSE44077. X-axis: sample type, Y-axis: lncRNA expression value (Mann–Whitney U test (M–W)); C. LINC00472 level in NSCLC cells (NCI-H1975, NCI-H157, NCI-H358, and NCI-H1299) and normal cells (BEAS-2B) was assayed using quantitative real-time polymerase chain reaction (qPCR) (Kruskal–Wallis test); D. Fluorescence in situ hybridization (FISH) (400×) was conducted to verify location of LINC00472 in NSCLC tissues. After the nuclei and cytoplasm of NCI-H1299 and NCI-H358 cells were separated, the expression of GAPDH (cytoplasmic marker), U6 (nuclear marker) and LINC00472 was assessed (M–W; **p < 0.01; ***p < 0.001). The horizontal lines represent the mediansFig. 1. LINC00472 is downregulated in non-small cell lung cancer (NSCLC) and predominantly located in the cytoplasm. A. Volcano map of differential genes in NSCLC gene expression chip GSE44077. X-axis represents the log10 p-value, while Y-axis represents the logFC value; the red points represent significantly upregulated genes in the tumor, the green points represent markedly downregulated genes in the tumor, and the black points represent genes with no significant difference; B. LINC00472 level in normal (left) and tumor (right) groups in GSE44077. X-axis: sample type, Y-axis: lncRNA expression value (Mann–Whitney U test (M–W)); C. LINC00472 level in NSCLC cells (NCI-H1975, NCI-H157, NCI-H358, and NCI-H1299) and normal cells (BEAS-2B) was assayed using quantitative real-time polymerase chain reaction (qPCR) (Kruskal–Wallis test); D. Fluorescence in situ hybridization (FISH) (400×) was conducted to verify location of LINC00472 in NSCLC tissues. After the nuclei and cytoplasm of NCI-H1299 and NCI-H358 cells were separated, the expression of GAPDH (cytoplasmic marker), U6 (nuclear marker) and LINC00472 was assessed (M–W; **p < 0.01; ***p < 0.001). The horizontal lines represent the medians

Article Snippet: The sequence of LINC00472 (full-length) was cloned using the SMARTerTM RACE cDNA kit (cat. No. 634858/59; Takara, Kusatsu, Japan).

Techniques: Gene Expression, Expressing, MANN-WHITNEY, Real-time Polymerase Chain Reaction, Fluorescence, In Situ Hybridization, Marker

Journal: Advances in clinical and experimental medicine : official organ Wroclaw Medical University

Article Title: LncRNA-LINC00472 suppresses the malignant progression of non-small cell lung cancer via modulation of the miRNA-1275/Homeobox A2 axis.

doi: 10.17219/acem/168431

Figure Lengend Snippet: Fig. 3. LINC00472 targets to inhibit miR-1275 level in NSCLC cells. A. Identification of the downstream miRNA of LINC00472. The left side is the prediction result of the RNA22 database, the right side is the differential

Article Snippet: The sequence of LINC00472 (full-length) was cloned using the SMARTerTM RACE cDNA kit (cat. No. 634858/59; Takara, Kusatsu, Japan).

Techniques:

Journal: Advances in clinical and experimental medicine : official organ Wroclaw Medical University

Article Title: LncRNA-LINC00472 suppresses the malignant progression of non-small cell lung cancer via modulation of the miRNA-1275/Homeobox A2 axis.

doi: 10.17219/acem/168431

Figure Lengend Snippet: Fig. 4. LINC00472 reverses the impact of miR-1275 on the malignant phenotype of non-small cell lung cancer (NSCLC) cells. A–C. Proliferation (A), migration and invasion (100×) (B), and apoptosis (C) of NCI-H1299 and

Article Snippet: The sequence of LINC00472 (full-length) was cloned using the SMARTerTM RACE cDNA kit (cat. No. 634858/59; Takara, Kusatsu, Japan).

Techniques: Migration

Journal: Advances in clinical and experimental medicine : official organ Wroclaw Medical University

Article Title: LncRNA-LINC00472 suppresses the malignant progression of non-small cell lung cancer via modulation of the miRNA-1275/Homeobox A2 axis.

doi: 10.17219/acem/168431

Figure Lengend Snippet: Fig. 5. LINC00472 promotes HOXA2 expression level and affects epithelial–mesenchymal transition (EMT), metastasis and apoptosis-related protein levels via inhibiting miR-1275. A. Identification of the downstream

Article Snippet: The sequence of LINC00472 (full-length) was cloned using the SMARTerTM RACE cDNA kit (cat. No. 634858/59; Takara, Kusatsu, Japan).

Techniques: Expressing

Journal: Advances in clinical and experimental medicine : official organ Wroclaw Medical University

Article Title: LncRNA-LINC00472 suppresses the malignant progression of non-small cell lung cancer via modulation of the miRNA-1275/Homeobox A2 axis.

doi: 10.17219/acem/168431

Figure Lengend Snippet: Fig. 6. LINC00472/miR-1275/HOXA2 axis regulates the progression of non-small cell lung cancer (NSCLC) cell phenotype. A. Proliferation of NCI-H1299 and NCI-H358 cells in each treatment group was assayed using Cell

Article Snippet: The sequence of LINC00472 (full-length) was cloned using the SMARTerTM RACE cDNA kit (cat. No. 634858/59; Takara, Kusatsu, Japan).

Techniques: