hpv18 positive human cervical cancer cell line hela  (ATCC)

Journal: bioRxiv

Article Title: Human Papillomavirus Infection Blunts STING Signaling and Alters Downstream Response

doi: 10.1101/2025.11.06.686957

Figure Lengend Snippet: Both oncogenes are needed to impact cGAMP levels. (A) western blot analysis for functional markers p53 and PTPN14, N= 3 using TTL system (B) cGAMP ELISA following DNA stimulation, N=4. (C) p-values for cGAMP ELISA of each cell line relative to HPV18 at specified time points. (D) Western blot showing function expression of E6 and/or E7 in overexpression cell lines. N=3 (E) cGAMP ELISA following DNA transfection, N=3 Statistics for (C and E) were calculated with two-way ANOVA, followed by Dunnett’s multiple comparison test. * p < 0.05; **p < 0.01; *** p < 0.001. Error bars indicate SEM.

Article Snippet: Primary human foreskin keratinocytes (HFKs) with or without HPV18 genomes were treated with 25 μg/mL 2′3′-cGAMP (InvivoGen tlrl-nacga23) to directly activate STING.

Techniques: Western Blot, Functional Assay, Enzyme-linked Immunosorbent Assay, Expressing, Over Expression, Transfection, Comparison

Journal: bioRxiv

Article Title: Human Papillomavirus Infection Blunts STING Signaling and Alters Downstream Response

doi: 10.1101/2025.11.06.686957

Figure Lengend Snippet: HPV18(+) cells produce more cGAMP than parental control cells. Quantification of cGAMP by ELISA following DNA stimulation of HPV(-) cells (black) and HPV18(+) cells (red). N=3 donors, 3 reps in each donor, average of the 9 replicates, (A) replicates separated by donor (B). Statistics for (A) were calculated using a linear mixed-effects model with “HPV” and “Time” as fixed effects and donor as a random effect to account for inter-donor variability. Statistics for (B) were calculated with a two-way ANOVA followed by a Dunnett’s post hoc test. * p<0.05; ** p<0.01; *** p<0.001Error bars for represent SEM, calculated by first averaging technical replicates (n=3 per donor) with propagated error, and then computing SEM across biological replicates (n=3 donors).

Article Snippet: Primary human foreskin keratinocytes (HFKs) with or without HPV18 genomes were treated with 25 μg/mL 2′3′-cGAMP (InvivoGen tlrl-nacga23) to directly activate STING.

Techniques: Control, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Human Papillomavirus Infection Blunts STING Signaling and Alters Downstream Response

doi: 10.1101/2025.11.06.686957

Figure Lengend Snippet: HPV decreases STING and IRF3 activation. (A) Western blot showing DNA-treated HPV18(+) and parental cells. Quantification of (B) pSTING, average of the 9 replicates (C) pSTING, separated by donor (D) pIRF3 western blots, average of the 9 replicates, (E) pIRF3, separated by donor. N=3 donors, 3 replicates in each. Statistics for the average of 9 (B and D) were calculated using a linear mixed-effects model with “HPV” and “Time” as fixed effects and donor as a random effect to account for inter-donor variability. Statistics for each cell line (C and E) were calculated with a two-way ANOVA followed by a Dunnett’s post hoc test. * p < 0.05; ** p<0.01; *** p<0.001. Error bars represent SEM, calculated by first averaging technical replicates (n=3 per donor) with propagated error, and then computing SEM across biological replicates (n=3 donors). Slope was calculated by fitting linear regressions within each time interval, and the values were subsequently analyzed using a linear mixed-effects model.

Article Snippet: Primary human foreskin keratinocytes (HFKs) with or without HPV18 genomes were treated with 25 μg/mL 2′3′-cGAMP (InvivoGen tlrl-nacga23) to directly activate STING.

Techniques: Activation Assay, Western Blot

Journal: bioRxiv

Article Title: Human Papillomavirus Infection Blunts STING Signaling and Alters Downstream Response

doi: 10.1101/2025.11.06.686957

Figure Lengend Snippet: Both oncogenes are needed to impact STING activation. (A) Western blot for total STING, pSTING, and αTubulin for the TTL cell lines, N=3. (B) Densitometry of pSTING normalized to αTubulin and a loading control lysate run on all blots. (C) p-values for pSTING densitometry of each cell line relative to HPV18 cells at specified time points. (D) Western blot for total STING, pSTING, and αTubulin for the over expression cell lines, N=3. (E) Densitometry of pSTING normalized to αTubulin and a loading control lysate run on all blots. (F) p-values for pSTING densitometry of each cell line relative to E6/E7 expressing cells at specified time points. Statistics for (C and F) were calculated with two-way ANOVA, followed by Dunnett’s multiple comparison test. * p < 0.05; ** p < 0.01; *** p < 0.001. Error bars indicate SEM.

Article Snippet: Primary human foreskin keratinocytes (HFKs) with or without HPV18 genomes were treated with 25 μg/mL 2′3′-cGAMP (InvivoGen tlrl-nacga23) to directly activate STING.

Techniques: Activation Assay, Western Blot, Control, Over Expression, Expressing, Comparison

Journal: eLife

Article Title: The long noncoding RNA lnc-FANCI-2 intrinsically restricts RAS signaling in human papillomavirus type 16-infected cervical cancer cells

doi: 10.7554/eLife.102681

Figure Lengend Snippet: ( A and B ) Expression of lnc-FANCI-2 (red) in HPV16 + cervical squamous cell carcinoma tissues was examined by RNAscope RNA in situ hybridization (RNA-ISH) analysis. Nuclei were stained with DAPI (blue). The corresponding regions ( A ) in H&E staining are shown for the adjacent region from the tumor nest and histotype. ( C ) Human foreskin keratinocyte (HFK)-derived raft cultures without (HFK) or with HPV16 or HPV18 infections (HFK-HPV16 or HFK-HPV18) were examined at day 10 for lnc-FANCI-2 RNA by RNAscope RNA-ISH analysis. Scale bars: 25 μm in the original figures and 10 μm in the zoomed insets.

Article Snippet: Cell line ( Homo sapiens ) , HeLa (HPV18 + ) , ATCC , CCL-2 , Cervical cancer cell line.

Techniques: Expressing, RNAscope, RNA In Situ Hybridization, Staining, Derivative Assay

Journal: eLife

Article Title: The long noncoding RNA lnc-FANCI-2 intrinsically restricts RAS signaling in human papillomavirus type 16-infected cervical cancer cells

doi: 10.7554/eLife.102681

Figure Lengend Snippet: ( A ) RT-qPCR detection of lnc-FANCI-2 in HPV16 + cervical cancer cell lines SiHa, CaSki, and W12 20861 (integrated HPV16) and 20863 (episomal HPV16), HPV18 + cervical cancer cell lines HeLa and C4II, and HPV - cell lines C33A (cervical cancer cells with mutations of p53 and pRb), HCT116 (colorectal cancer cells), BCBL-1 (body cavity B lymphoma cells), HEK293 (Ad5 E1/E2-immortalized human kidney cells), and HaCaT (spontaneously immortalized human epidermal cells) in triplicates. ( B ) HeLa cells express no lnc-FANCI-2 when compared with C33A cells by northern blot. ( C ) lnc-FANCI-2 is mainly cytoplasmic in CaSki but nuclear in SiHa and C33A cells. Cytoplasmic and nuclear fractionation efficiency was blotted for nuclear SRSF3 (serine- and arginine-rich splicing factor 3) and cytoplasmic GAPDH. Total fractionated cytoplasmic and nuclear RNAs were quantified for lnc-FANCI-2 by RT-qPCR in triplicates, with GAPDH RNA serving as an internal control for RNA fractionation efficiency. ( D ) Subcellular lnc-FANCI-2 (red) localization in CaSki, SiHa, and C33A cells determined by RNAscope RNA in situ hybridization (RNA-ISH) analysis. Nuclei were stained with DAPI (blue). Scale bars: 25 μm in the top and 10 μm in the zoom. Figure 2—source data 1. Northern blot and western blot data for . Figure 2—source data 2. Northern blot and western blot data for .

Article Snippet: Cell line ( Homo sapiens ) , HeLa (HPV18 + ) , ATCC , CCL-2 , Cervical cancer cell line.

Techniques: Quantitative RT-PCR, Northern Blot, Fractionation, Control, RNAscope, RNA In Situ Hybridization, Staining, Western Blot

Journal: eLife

Article Title: The long noncoding RNA lnc-FANCI-2 intrinsically restricts RAS signaling in human papillomavirus type 16-infected cervical cancer cells

doi: 10.7554/eLife.102681

Figure Lengend Snippet: ( A and B ) Expression of lnc-FANCI-2 (red) in HPV16 + cervical squamous cell carcinoma tissues was examined by RNAscope RNA in situ hybridization (RNA-ISH) analysis. Nuclei were stained with DAPI (blue). The corresponding regions ( A ) in H&E staining are shown for the adjacent region from the tumor nest and histotype. ( C ) Human foreskin keratinocyte (HFK)-derived raft cultures without (HFK) or with HPV16 or HPV18 infections (HFK-HPV16 or HFK-HPV18) were examined at day 10 for lnc-FANCI-2 RNA by RNAscope RNA-ISH analysis. Scale bars: 25 μm in the original figures and 10 μm in the zoomed insets.

Article Snippet: Cell line ( Homo sapiens ) , C4II (HPV18 + ) , ATCC , CRL-1595 , Cervical cancer cell line.

Techniques: Expressing, RNAscope, RNA In Situ Hybridization, Staining, Derivative Assay

Journal: eLife

Article Title: The long noncoding RNA lnc-FANCI-2 intrinsically restricts RAS signaling in human papillomavirus type 16-infected cervical cancer cells

doi: 10.7554/eLife.102681

Figure Lengend Snippet: ( A ) RT-qPCR detection of lnc-FANCI-2 in HPV16 + cervical cancer cell lines SiHa, CaSki, and W12 20861 (integrated HPV16) and 20863 (episomal HPV16), HPV18 + cervical cancer cell lines HeLa and C4II, and HPV - cell lines C33A (cervical cancer cells with mutations of p53 and pRb), HCT116 (colorectal cancer cells), BCBL-1 (body cavity B lymphoma cells), HEK293 (Ad5 E1/E2-immortalized human kidney cells), and HaCaT (spontaneously immortalized human epidermal cells) in triplicates. ( B ) HeLa cells express no lnc-FANCI-2 when compared with C33A cells by northern blot. ( C ) lnc-FANCI-2 is mainly cytoplasmic in CaSki but nuclear in SiHa and C33A cells. Cytoplasmic and nuclear fractionation efficiency was blotted for nuclear SRSF3 (serine- and arginine-rich splicing factor 3) and cytoplasmic GAPDH. Total fractionated cytoplasmic and nuclear RNAs were quantified for lnc-FANCI-2 by RT-qPCR in triplicates, with GAPDH RNA serving as an internal control for RNA fractionation efficiency. ( D ) Subcellular lnc-FANCI-2 (red) localization in CaSki, SiHa, and C33A cells determined by RNAscope RNA in situ hybridization (RNA-ISH) analysis. Nuclei were stained with DAPI (blue). Scale bars: 25 μm in the top and 10 μm in the zoom. Figure 2—source data 1. Northern blot and western blot data for . Figure 2—source data 2. Northern blot and western blot data for .

Article Snippet: Cell line ( Homo sapiens ) , C4II (HPV18 + ) , ATCC , CRL-1595 , Cervical cancer cell line.

Techniques: Quantitative RT-PCR, Northern Blot, Fractionation, Control, RNAscope, RNA In Situ Hybridization, Staining, Western Blot