gene exp itga1 hs00235006 m1 (Thermo Fisher)
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gene exp itga1 hs00235006 m1
Gene Exp Itga1 Hs00235006 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/transcriptome+sequencing+experiment/Gene+Exp%2E+ITGA1%2C+Hs00235006_m1/pmc07458291-215-13-14
Average 98 stars, based on 1 article reviews
Gene Exp Itga1 Hs00235006 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/transcriptome+sequencing+experiment/Gene+Exp%2E+ITGA1%2C+Hs00235006_m1/pmc07458291-215-13-14
Average 98 stars, based on 1 article reviews
gene exp itga1 hs00235006 m1 - by Bioz Stars,
2026-10
98/100 stars
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1) Product Images from "Human papillomavirus type 38 alters wild-type p53 activity to promote cell proliferation via the downregulation of integrin alpha 1 expression"
Article Title: Human papillomavirus type 38 alters wild-type p53 activity to promote cell proliferation via the downregulation of integrin alpha 1 expression
Journal: PLoS Pathogens
doi: 10.1371/journal.ppat.1008792
Figure Legend Snippet: (A) Primary HKs were transduced with pLXSN HPV38 E6/E7 or pLSXN. mRNA levels were measured by RT-qPCR and normalized to GAPDH . Error bars represent standard deviations from 3 biological replicates of 2 different donors ( n = 6). ***, p <0.001; ****, p <0.0001; ns, not significant. (B) Total RNA levels of HKs expressing or not expressing HPV38 E6 and E7 were analyzed by TaqMan PCR. Commercial probes for ITGA1 and GAPDH were used. Results were normalized to GAPDH . Data shown are the means of 3 independent experiments for 2 different donors ( n = 6). ****, p <0.0001. (C) Proteins extracts from HKs expressing or not expressing HPV38 E6 and E7 were analyzed by immunoblotting (IB) with the indicated antibodies. (D) The TaqMan assay was also performed as previously described in primary HKs previously retrovirally transduced with the hTERT gene and expressing E6 and/or E7 from HPV38 ( n = 3). Results were normalized to GAPDH . ****, p <0.0001. (E) Proteins extracts from hTERT pLXSN or hTERT HPV38 E6/E7 cells were analyzed by IB with the indicated antibodies. Images shown are representative examples of 2 different experiments. (F) hTERT pLXSN or hTERT HPV38 E6/E7 cells were plated on coverslips and after 24 h were probed for ITGA1 using anti-ITGA1 antibody followed by secondary Alexa Fluor 488-conjugated antibody. Nuclei were stained with DAPI (pseudocoloured red), and cells were analyzed under a microscope. Images were merged using ImageJ software.
Techniques Used: Transduction, Quantitative RT-PCR, Expressing, Western Blot, TaqMan Assay, Staining, Microscopy, Software
Figure Legend Snippet: (A) Electromobility shift assay performed with 38HK nuclear protein extracts and biotinylated probes containing p53RE WT or mutated sequences. Probes were incubated and cross-linked with protein extracts. Unlabeled WT or mutant p53RE2 probes were used as a control. Images shown are representative examples of 2 different experiments. (B) 38HK were cross-linked and chromatin was processed for ChIP using p53 antibody. Results were analyzed by qPCR with primers spanning p53RE1, p53RE2, p53RE3, or the intergenic region of chromosome 22 as a negative control (nc). Error bars represent standard deviations of 3 independent experiments performed in triplicate. **, p <0.01. (C) HKs or 38HK were cross-linked and chromatin was processed for ChIP using p53 or IgG antibodies. Results were analyzed by qPCR using primers spanning for p53 REs of the ITGA1 promoter and normalized to IgG enrichment (negative control). Error bars represent the standard deviation of 2 independent experiments performed in 2 different HKs donors. *, p <0.05, ns, not significant. (D) Cell lysate was incubated with WT biotinylated probe containing p53 REs of the ITGA1 promoter. Incubation without a probe was used as a control. DNA-associated proteins were recovered by precipitation with streptavidin beads and analyzed by IB. Images shown are representative examples of 3 independent experiments. Signals of 3 different IBs were quantified with Image Lab software (right panel). Data shown are the means of 3 independent experiments. *, p <0.05. (E) Chromatin from 38HK was processed for ChIP experiments using p53 or DNMT1 antibodies. Results were obtained by qPCR with primers spanning p53RE2 or the intergenic region of chromosome 22 (nc). Error bars indicate standard deviations from 3 independent experiments performed in duplicate. **, p <0.01; ***, p <0.001. (F) 38HK were cultured in medium containing cyclic pifithrin-α hydrobromide or DMSO as a control. Chromatin was processed for ChIP using p53 or DNMT1 antibodies. Results were obtained by qPCR using primers spanning p53RE2. Data shown are the means of 2 independent experiments performed in triplicate. *, p <0.05, **, p <0.01. (G) Chromatin was processed for a ChIP-reChIP assay in which p53-immunoprecipitated DNA was re-immunoprecipitated by DNMT1. Enrichment of p53RE2 or the intergenic region of chromosome 22 (nc) was obtained by qPCR. Data shown are the means of 3 independent experiments performed in triplicate. **, p <0.01. (H) Nuclear protein extracts from 38HK were processed for IP. Agarose beads were conjugated with IgG or p53 antibodies. Conjugated beads were incubated with protein lysate overnight. IgG was used as a control. Results were obtained by IB using the indicated antibodies. (I) 38HK were transfected with DNMT1 siRNA or control siRNA (Scramble). After 72 h, a ChIP assay was performed with p53 or DNMT1 antibodies. Results were obtained by qPCR using p53RE2 primers. Error bars represent standard deviations from 3 independent experiments. *, p <0.05; **, p <0.01. DNMT1 protein levels in different cells were determined by IB with the indicated antibodies (right panel).
Techniques Used: Electro Mobility Shift Assay, Incubation, Mutagenesis, Control, Negative Control, Standard Deviation, Software, Cell Culture, Immunoprecipitation, Transfection
Figure Legend Snippet: (A) mRNA levels of 38HK treated with cyclic pifithrin-α hydrobromide or DMSO for 6 h were analyzed by RT-qPCR and normalized to GAPDH . Histograms represent the mean of at least 3 independent experiments. ****, p <0.0001. (B and C) ITGA1 and p53 mRNA and protein levels from 38HK expressing WT p53 (Scramble) or with CRISPR/Cas9-mediated p53 deletion (CRISPR-p53) were measured by RT-qPCR (B) and IB (C). (B) Gene expression was normalized to GAPDH . (C) Protein quantification was normalized to β-actin. Signals of 4 different IBs were quantified with Image Lab software (right panel). Data shown are the means of 4 independent experiments. *, p <0.05. (D and E) 38HK N-HA-p53 or p53-C-HA cells were generated by retroviral transduction with WT p53 tagged at the N- or C-terminus. As a control, 38HK were transduced with the corresponding empty plasmid. Protein extracts and total mRNA levels were processed for IB and RT-qPCR analysis, respectively. (D) IB images shown are representative examples of 3 independent experiments. (E) ITGA1 mRNA levels were normalized to GAPDH . Error bars indicate standard deviations of 4 independent experiments. ***, p <0.001. (F) 38HK were transfected with control siRNA (Scramble) or with DNMT1 siRNA (siDNMT1). After 72 h, cells were collected for RNA extraction and RT-qPCR analysis. Error bars indicate standard deviations of 3 independent experiments. *, p <0.05; ***, p <0.001. (G) ITGA1 expression was evaluated by RT-qPCR after 24 h of treatment with 5-aza-2′- deoxycytidine (Aza) or DMSO at 30 μM final concentration. Error bars represent standard deviations of 3 independent experiments. **, p <0.01. (H) H3K9ac at the ITGA1 promoter was evaluated by ChIP assay after treatment with Aza or DMSO as previously described ( n = 4). Results were obtained by qPCR using primers for p53RE2. *, p <0.05.
Techniques Used: Quantitative RT-PCR, Expressing, CRISPR, Gene Expression, Software, Generated, Retroviral, Transduction, Control, Plasmid Preparation, Transfection, RNA Extraction, Concentration Assay
Figure Legend Snippet: (A) Protein extracts from 38HK were processed for oligonucleotide pulldown as previously described. Images shown are representative examples of 3 independent experiments. (B) HK and 38HK were processed for protein extraction and IB with the indicated antibodies. After incubation with p446PKR antibody, the membrane was stripped and incubated with total PKR antibody. (C) Proteins extracts from U2OS cells retrovirally transduced with E6 and/or E7 from HPV38 were analyzed by IB with the indicated antibodies. (D) 38HK were treated with PKR inhibitor, 2AP, or PBS:glacial acetic acid (200:1) as a control for 4 h at 10 mM final concentration. p-p53 S392 and p53 band intensities were quantified and normalized to total p53 (central panel) or β-actin (right panel). Membranes were first incubated with p446PKR, then stripped and incubated with total PKR. Data shown are the means of 3 independent experiments. *, p <0.05, **, p <0.01. (E and F) 38HK were treated with 2AP, and ITGA1 mRNA (D) and protein levels (E) were determined by RT-PCR and IB, respectively. (D) Data shown are the means of 3 independent experiments (**, p <0.01). (E) Images shown are representative examples of 3 independent experiments. (G and H) ChIP assay using p53 or DNMT1 antibodies was performed in 38HK treated with or PBS:glacial acetic acid (200:1) as a control (CTR) or 2AP. Data shown are the means of 2 independent experiments performed in duplicate by qPCR using p53RE2 primers. *, p <0.05.
Techniques Used: Protein Extraction, Incubation, Membrane, Transduction, Control, Concentration Assay, Reverse Transcription Polymerase Chain Reaction
Figure Legend Snippet: (A) 38HK were transfected with ITGA1 cDNA (ITGA1) or empty plasmid control (pcDNA). After zeocin selection, 38HK were fixed with crystal violet (left panel) and total colony number per well was counted (right panel). Data shown are the means of 3 independent experiments. ****, p <0.0001. (B) 38HK transfected with ITGA1 cDNA (ITGA1) or empty plasmid control (pcDNA) were fixed and stained with propidium iodide for flow cytometry analysis. The histograms (right) represent the means of the sub-G0 population of 3 independent experiments. **, p <0.01. (C and D) Total protein and mRNA extracts from transfected 38HK were analyzed by IB and RT-qPCR. (C) Protein band intensity was quantified and normalized to β-actin. (D) ITGA1 , EGFR , and CCND1 mRNA levels were normalized to GAPDH . For (C) and (D), data shown are the means of 4 independent experiments *, p <0.05; **, p <0.01; ***, p <0.001; ns, not significant.
Techniques Used: Transfection, Plasmid Preparation, Control, Selection, Staining, Flow Cytometry, Quantitative RT-PCR
Figure Legend Snippet: (A) Skin keratinocytes were isolated from WT animals ( n = 4) and K14 HPV38 E6/E7 transgenic mice ( n = 3). After 30 weeks of UV irradiation, cSCC samples (cSCC1–3) were isolated from HPV38 E/E7 transgenic mice. Total RNA extraction was performed and Itga1 mRNA levels were determined by quantitative RT-PCR by normalizing to Gapdh . Whole-exome sequencing of the same mice was also performed. Mutational analysis of the Trp53 , Itga1 , and Egfr genes was performed as described in Materials and Methods. (B) Genomic position of the exonic mutations and the corresponding amino acid change are represented for the Itga1 and Egfr genes. WT exons are represented as blue boxes, and mutated exons are represented in red boxes. Text boxes describe the cSCC sample, the genomic position of the nucleotide change on the GRCm38/mm10 mouse reference genome, the type of mutation, and the corresponding amino acid change.
Techniques Used: Isolation, Transgenic Assay, Irradiation, RNA Extraction, Quantitative RT-PCR, Sequencing, Mutagenesis
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