Journal: International journal of cancer

Article Title: Autocrine/paracrine erythropoietin signalling promotes JAK/STAT-dependent proliferation of human cervical cancer cells.

doi: 10.1002/ijc.25935

Figure Lengend Snippet: Figure 3. Exogenous Epo induces proliferation of cervical cancer cells. SiHa, HeLa and C33 cells were treated with increasing

Article Snippet: Cervical cancer-derived HeLa, SiHa and C33 cells (American Type Culture Collection, Manassas VA) were maintained in Dulbecco’s modified Eagle’s medium (DMEM; Invitro, Mexico DF, Mexico) supplemented with 2% heat-inactivated foetal bovine serum (FBS) (Invitrogen, Carlsbad, CA).

Techniques:

Journal: bioRxiv

Article Title: Transcriptional Regulation of the P-bodies Associated Psoriasis Candidate Gene CCHCR1

doi: 10.1101/2023.04.08.535951

Figure Lengend Snippet: ( a ) RT-qPCR for the expression of CCHCR1 and TCF19 in HeLa cells transfected with E2F activators (E2F1, E2F2 or E2F3). pEGFP-N1 was the empty vector control. Gene expression level was normalized to the expression level of RPL13A . Results were mean ± SD of three replicates. Statistically significant differences of the gene expression levels were compared to empty vector control and were analyzed by paired t-test (** = P ≤ 0.01). ( b ) Down-regulation of CCHCR1 and TCF19 mRNA by E2F1 knockdown. Cells were transfected with siE2F1 and were harvested for the time indicated (time post-transfection). mRNA expressions (normalized to RPL13A gene expression) were analyzed by RT-qPCR. Results were mean ± SD of three replicates. Statistically significant differences of the gene expression levels were compared to 0 h post-siE2F1 transfection and were analyzed by paired t-test (*** = P ≤ 0.001).

Article Snippet: Human cervical cancer HeLa cells (ATCC ® CRM-CCL-2 TM ) were cultured in DMEM supplemented with 10% fetal bovine serum (FBS) and maintained at 37°C in a humidified atmosphere of 5% CO 2 .

Techniques: Quantitative RT-PCR, Expressing, Transfection, Plasmid Preparation, Control, Gene Expression, Knockdown

Journal: bioRxiv

Article Title: Transcriptional Regulation of the P-bodies Associated Psoriasis Candidate Gene CCHCR1

doi: 10.1101/2023.04.08.535951

Figure Lengend Snippet: ( a ) HeLa cells were synchronized and arrested in G1/S transition by a double thymidine block. The cells were released and collected at different time points, stained with propidium iodide (PI). The percentage of cells in different cell cycle phases in each time point (G1/S, S, G2/M, and G1) were analyzed by flow cytometry. ( b ) Percentage of cells in different cell cycles after double thymidine block and release. (C) Relative expression of CCHCR1 and TCF19 in different cell cycle phases. CCHCR1 and TCF19 gene expression were normalized to RPL13A gene expression. The time points after release from the double thymidine block were indicated for different cell cycle phases; i.e. 0 h: G1/S, 2 h: S, 8 h: G2/M and 14 h: G1. Results were mean ± SD of three replicates. Statistically significant differences of the gene expressions were compared to 0 h after double thymidine release (G1/S transition) and were analyzed by paired t-test (** = P ≤ 0.01 and *** = P ≤ 0.001)

Article Snippet: Human cervical cancer HeLa cells (ATCC ® CRM-CCL-2 TM ) were cultured in DMEM supplemented with 10% fetal bovine serum (FBS) and maintained at 37°C in a humidified atmosphere of 5% CO 2 .

Techniques: Blocking Assay, Staining, Flow Cytometry, Expressing, Gene Expression

Journal: bioRxiv

Article Title: Transcriptional Regulation of the P-bodies Associated Psoriasis Candidate Gene CCHCR1

doi: 10.1101/2023.04.08.535951

Figure Lengend Snippet: ( a ) HeLa cells were transfected with the corresponding siRNAs. Non-viable cells were counted using trypan blue exclusion assay. siCTL was Silencer select negative control #1 (Ambion, Life Technologies, USA). Results are mean ± SD of five replicates. Statistically significant differences of the cell counts were compared to the untransfected samples in the corresponding time point, and were analyzed by paired t-test (* = ≤0.05 and ** = P ≤ 0.01). ( b ) Up-regulation of p21 mRNA was observed after knockdown of CCHCR1 . siRNAs were transfected into HeLa cells and were collected for analysis 4 days post-transfection. Gene expressions (normalized to RPL13A gene expression) were analyzed by RT-qPCR. Results are mean ± SD of three replicates. Statistically significant differences of the gene expressions were compared to untransfected sample and were analyzed by paired t-test (* = ≤0.05, ** = P ≤ 0.01 and *** = P ≤ 0.001).

Article Snippet: Human cervical cancer HeLa cells (ATCC ® CRM-CCL-2 TM ) were cultured in DMEM supplemented with 10% fetal bovine serum (FBS) and maintained at 37°C in a humidified atmosphere of 5% CO 2 .

Techniques: Transfection, Trypan Blue Exclusion Assay, Negative Control, Knockdown, Gene Expression, Quantitative RT-PCR

Journal: bioRxiv

Article Title: Transcriptional Regulation of the P-bodies Associated Psoriasis Candidate Gene CCHCR1

doi: 10.1101/2023.04.08.535951

Figure Lengend Snippet: ( a ) HeLa cells were treated with 0.1% DMSO (vehicle control) or 2 μM CPT for 24 hours. The percentage of cells in different cell cycle phases were analyzed by flow cytometry. ( b ) Relative expression of CCHCR1 and TCF19 (normalized to RPL13A gene expression) in HeLa cells after the treatment with CPT for 24 hours. Results are mean ± SD of three replicates. Statistically significant differences of the gene expression level were compared to 0.1% DMSO treatment and were analyzed by paired t-test (* = ≤0.05 and ** = P ≤ 0.01).

Article Snippet: Human cervical cancer HeLa cells (ATCC ® CRM-CCL-2 TM ) were cultured in DMEM supplemented with 10% fetal bovine serum (FBS) and maintained at 37°C in a humidified atmosphere of 5% CO 2 .

Techniques: Control, Flow Cytometry, Expressing, Gene Expression