research cell line source s human osteosarcoma u2os cells  (ATCC)

Journal: bioRxiv

Article Title: Comparative analysis of wavelength-specific UV stress granule formation

doi: 10.64898/2026.03.15.711948

Figure Lengend Snippet: Representative images of SG formation by G3BP1 and TIA1 signals in wt U2OS cells treated as indicated. Cells were either untreated, treated with 250μM As III for 1h, or exposed to 15mJ/cm 2 of UV. UV samples were harvested and processed for imaging 4 hours after exposure. Yellow arrows indicate SGs. Scale bar = 20μm.

Article Snippet: Cultured cell lines U2OS (ATCC HTB-96), wild-type C57-Bl6 MEF (primary MEFs transformed by the 3T3 method, a gift of Joel D. Richter, UMass Chan Medical School) and HaCaT, were maintained at 37oC in a CO2 incubator (5% CO2) in DMEM (Gibco) supplemented with 10% FBS, 1% penicillin/streptomycin and 1% glutamine (0.2% glutamine for HaCaT cells).

Techniques: Imaging

Journal: bioRxiv

Article Title: Comparative analysis of wavelength-specific UV stress granule formation

doi: 10.64898/2026.03.15.711948

Figure Lengend Snippet: (A) Representative images of SG formation by G3BP1 and TIA1 signals in HaCaT cells treated as indicated. Cells were either untreated, treated with 250μM As III for 1h, or exposed to 15mJ/cm 2 of UV. UV samples were harvested and processed for imaging 4 hours after exposure. (B) Quantification of SG formation by G3BP1 and TIA1 signals in wt U2OS, HaCaT, and wt MEF cells treated as indicated. Cells were either untreated, treated with 250μM As III for 1h, or exposed to 15mJ/cm 2 of UV. UV samples were harvested and processed for imaging 4 hours after exposure. At least 250 cells and 3 fields of view were scored to calculate percent SG formation. n =3; error bars are ±SD; ns P > 0.05, ** P ≤ 0.01, **** P ≤ 0.0001 by one-way ANOVA and Dunnett’s post-hoc test. (C) Quantification of SG formation by G3BP1 and TIA1 signals in wt U2OS cells treated as indicated. Cells were either untreated, treated with 250μM As III for 1h, or exposed to 150mJ/cm 2 of UV. UV samples were harvested and processed for imaging 4 hours after exposure. At least 250 cells and 3 fields of view were scored to calculate percent SG formation. n =3; error bars are ±SD; ns P > 0.05, **** P ≤ 0.0001 by one-way ANOVA and Dunnett’s post-hoc test. Scale bar = 20μm.

Article Snippet: Cultured cell lines U2OS (ATCC HTB-96), wild-type C57-Bl6 MEF (primary MEFs transformed by the 3T3 method, a gift of Joel D. Richter, UMass Chan Medical School) and HaCaT, were maintained at 37oC in a CO2 incubator (5% CO2) in DMEM (Gibco) supplemented with 10% FBS, 1% penicillin/streptomycin and 1% glutamine (0.2% glutamine for HaCaT cells).

Techniques: Imaging

Journal: bioRxiv

Article Title: Comparative analysis of wavelength-specific UV stress granule formation

doi: 10.64898/2026.03.15.711948

Figure Lengend Snippet: (A-D) Representative images of SGs identified by G3BP1 co-stained with relevant SG markers (A) DHX9, (B) Rps6, (C) eIF3n, (D) Poly(A) fluorescence in situ hybridization (FISH). wt U2OS cells were either untreated, treated with 250μM As III for 1h, or exposed to 150mJ/cm 2 of UV. UV samples were harvested and processed for imaging 4 hours after exposure. Cells were stained with relevant SG marker antibodies and observed and imaged under 63X magnification. Scale bar = 20μm.

Article Snippet: Cultured cell lines U2OS (ATCC HTB-96), wild-type C57-Bl6 MEF (primary MEFs transformed by the 3T3 method, a gift of Joel D. Richter, UMass Chan Medical School) and HaCaT, were maintained at 37oC in a CO2 incubator (5% CO2) in DMEM (Gibco) supplemented with 10% FBS, 1% penicillin/streptomycin and 1% glutamine (0.2% glutamine for HaCaT cells).

Techniques: Staining, Fluorescence, In Situ Hybridization, Imaging, Marker

Journal: bioRxiv

Article Title: Comparative analysis of wavelength-specific UV stress granule formation

doi: 10.64898/2026.03.15.711948

Figure Lengend Snippet: Representative images of wt U2OS cells treated as indicated and stained for G3BP1 and Geminin. Cells were either untreated, treated with 250μM As III for 1h, or exposed to 150mJ/cm 2 of UV. UV samples were harvested and processed for imaging 4 hours after exposure. Yellow arrows indicate cells in S, G2, or M phase. Red arrows indicate cells in G1 phase. Scale bar = 20μm.

Article Snippet: Cultured cell lines U2OS (ATCC HTB-96), wild-type C57-Bl6 MEF (primary MEFs transformed by the 3T3 method, a gift of Joel D. Richter, UMass Chan Medical School) and HaCaT, were maintained at 37oC in a CO2 incubator (5% CO2) in DMEM (Gibco) supplemented with 10% FBS, 1% penicillin/streptomycin and 1% glutamine (0.2% glutamine for HaCaT cells).

Techniques: Staining, Imaging

Journal: bioRxiv

Article Title: Comparative analysis of wavelength-specific UV stress granule formation

doi: 10.64898/2026.03.15.711948

Figure Lengend Snippet: (A-B) Representative images (A) and quantification (B) of SG formation in wt U2OS cells after treatment as indicated. Cells were either untreated or pre-treated with 4sU for 1h. Following pretreatment cells were either untreated, treated with 250μM As III for 1h, or treated with UVA. UV samples were harvested and processed for imaging 8 hours after exposure. At least 250 cells and 3 fields of view were scored to calculate percent SG formation. n =4; error bars are ±SD; ns P > 0.05, * P≤0.05 by unpaired t test. Scale bar = 20μm.

Article Snippet: Cultured cell lines U2OS (ATCC HTB-96), wild-type C57-Bl6 MEF (primary MEFs transformed by the 3T3 method, a gift of Joel D. Richter, UMass Chan Medical School) and HaCaT, were maintained at 37oC in a CO2 incubator (5% CO2) in DMEM (Gibco) supplemented with 10% FBS, 1% penicillin/streptomycin and 1% glutamine (0.2% glutamine for HaCaT cells).

Techniques: Imaging

Journal: bioRxiv

Article Title: Comparative analysis of wavelength-specific UV stress granule formation

doi: 10.64898/2026.03.15.711948

Figure Lengend Snippet: (A) Representative images of wt U2OS cells stained for G3BP1 and keratin 8 treated as indicated. Cells were either untreated, treated with 250μM As III for 1h, or exposed to 150mJ/cm 2 of UV. UV samples were harvested and processed for imaging 4 hours after exposure. (B) Representative images of wt U2OS cells co-transfected with keratin 8 and keratin18_mCherry. Cells were either untreated, treated with 250μM As III for 1h, or exposed to 150mJ/cm 2 of UV. UV samples were harvested and processed for imaging 4 hours after exposure. (C) Quantification of SG formation in untransfected and co-transfected cells by G3BP1 staining. Cells were treated as indicated. At least 200 cells and 3 fields of view were scored to calculate percent SG formation. n =3; error bars are ±SD; ns P > 0.05, ** P ≤ 0.01, *** P ≤ 0.001 by unpaired t -test. Scale bar = 20μm.

Article Snippet: Cultured cell lines U2OS (ATCC HTB-96), wild-type C57-Bl6 MEF (primary MEFs transformed by the 3T3 method, a gift of Joel D. Richter, UMass Chan Medical School) and HaCaT, were maintained at 37oC in a CO2 incubator (5% CO2) in DMEM (Gibco) supplemented with 10% FBS, 1% penicillin/streptomycin and 1% glutamine (0.2% glutamine for HaCaT cells).

Techniques: Staining, Imaging, Transfection

Journal: bioRxiv

Article Title: Comparative analysis of wavelength-specific UV stress granule formation

doi: 10.64898/2026.03.15.711948

Figure Lengend Snippet: (A-B) Representative images (A) and quantification of SG formation in U2OS cells after treated as indicated. Cells were either untreated, treated with NAC (10mM) for 1h, treated with 250μM As III for 1h, co-treated with NAC and 250μM As III for 1h, treated with UV, or co-treated with UV and NAC. UV samples were harvested and processed for imaging 4 hours after exposure. At least 250 cells and 3 fields of view were scored to calculate percent SG formation. n =3; error bars are ±SD; ns P > 0.05, ** P ≤ 0.01, **** P ≤ 0.0001 by unpaired t -test.

Article Snippet: Cultured cell lines U2OS (ATCC HTB-96), wild-type C57-Bl6 MEF (primary MEFs transformed by the 3T3 method, a gift of Joel D. Richter, UMass Chan Medical School) and HaCaT, were maintained at 37oC in a CO2 incubator (5% CO2) in DMEM (Gibco) supplemented with 10% FBS, 1% penicillin/streptomycin and 1% glutamine (0.2% glutamine for HaCaT cells).

Techniques: Imaging

Journal: Science Advances

Article Title: RAD51C-XRCC3 complex regulates FANCM-mediated R-loop resolution to safeguard genome integrity

doi: 10.1126/sciadv.aea5932

Figure Lengend Snippet: ( A ) Quantification of S9.6 staining in control, C/X3/X2-depleted U2OS cells with or without HU (1 mM, 4 hours) treatment. Data are represented as a box plot ( n = 3, two-tailed, unpaired t test with Welch’s correction on means of biological replicates, ≥242 cells were analyzed per condition). ( B ) Quantification of S9.6 staining in control, C/X3/X2-depleted U2OS cells with or without Aph (0.4 μM, 16 hours) treatment. Data are represented as a box plot ( n = 3, two-tailed, unpaired t test with Welch’s correction on means of biological replicates, ≥211 cells were analyzed per condition). ( C ) Quantification of γH2AX intensity in the presence or absence of HU (1 mM, 4 hours) treatment in control, C/X3-depleted cells with or without overexpression of RNH1-Flag. Data are represented as a scatter plot. Black lines indicate mean values ( n = 3, two-tailed unpaired t test, ≥241 cells were analyzed per condition). ( D ) Effect of RAD51C or XRCC3 depletion on fork progression/fork symmetry following 50 nM CPT treatment as indicated in the schematic. Top: Experimental schematic; middle: representative images; and bottom: quantification of sister fork ratio. Sister fork ratios were measured by dividing the longer sister fork by the shorter fork (IdU tracts). Data are represented as a scatter plot. Black lines indicate mean values ( n = 3, two-tailed unpaired t test on means of biological replicates, ≥40 sister fork pairs were analyzed per condition). P values are indicated.

Article Snippet: Human cell lines U2OS (American Type Culture Collection; HTB-96; RRID: CVCL_0042), HeLa Kyoto (Sachin Kotak laboratory; RRID: CVCL_1922), and hTERT RPE-1 (Sachin Kotak laboratory) were grown in Dulbecco’s modified Eagle medium supplemented with 10% fetal bovine serum, 1% penicillin/streptomycin (Sigma-Aldrich), and 1% Glutamax (Gibco) under standard conditions (5% CO 2 , humidified atmosphere) at 37°C.

Techniques: Staining, Control, Two Tailed Test, Over Expression

Journal: Science Advances

Article Title: RAD51C-XRCC3 complex regulates FANCM-mediated R-loop resolution to safeguard genome integrity

doi: 10.1126/sciadv.aea5932

Figure Lengend Snippet: ( A ) Representative immunoblot indicating levels of WT and R258H RAD51C-Flag after depletion of endogenous RAD51C in U2OS cells. MCM3 levels are shown as loading control. ( B ) Quantification of S9.6 staining after transfection of cells with the indicated DNA constructs, as shown in (A). Relative nuclear S9.6 intensity data are plotted as a box plot ( n = 3, two-tailed, unpaired t test with Welch’s correction on means of biological replicates, ≥252 cells were analyzed per condition). ( C ) Quantification of RNAPIIS2P + PCNA PLA foci after transfection of cells with the indicated DNA constructs. Data are represented as a scatter plot. Black lines indicate mean values ( n = 3, two-tailed unpaired t test on means of biological replicates, ≥303 cells were analyzed per condition). ( D ) Representative immunofluorescence images (top) and quantification (bottom) of S9.6 + Flag PLA foci in WT and R258H Flag-RAD51C–transfected cells. Data are represented as a scatter plot. Black lines indicate mean values ( n = 3, two-tailed unpaired t test on means of biological replicates, ≥731 cells were analyzed per condition). Scale bar, 5 μm. ( E ) Co-IP of exogenously expressed Flag-tagged WT and R258H RAD51C, followed by immunoblotting with indicated antibodies. Endogenous protein levels were shown in the input. ( F and G ) Representative immunofluorescence images (F) and quantification (G) of S9.6 + FANCM PLA foci in cells transfected with the indicated DNA constructs. Data are represented as a scatter plot. Black lines indicate mean values ( n = 3, two-tailed unpaired t test on means of biological replicates, ≥251 cells were analyzed per condition). Scale bar, 5 μm. P values are indicated. res, shRNA-resistant.

Article Snippet: Human cell lines U2OS (American Type Culture Collection; HTB-96; RRID: CVCL_0042), HeLa Kyoto (Sachin Kotak laboratory; RRID: CVCL_1922), and hTERT RPE-1 (Sachin Kotak laboratory) were grown in Dulbecco’s modified Eagle medium supplemented with 10% fetal bovine serum, 1% penicillin/streptomycin (Sigma-Aldrich), and 1% Glutamax (Gibco) under standard conditions (5% CO 2 , humidified atmosphere) at 37°C.

Techniques: Western Blot, Control, Staining, Transfection, Construct, Two Tailed Test, Immunofluorescence, Co-Immunoprecipitation Assay, shRNA

Journal: Science Advances

Article Title: RAD51C-XRCC3 complex regulates FANCM-mediated R-loop resolution to safeguard genome integrity

doi: 10.1126/sciadv.aea5932

Figure Lengend Snippet: ( A ) Experimental schematic and representative images, and ( B ) quantification of IdU/CldU ratio for fork protection assay in cells expressing the indicated mutants of RAD51C. Data are represented as a scatter plot. Black lines indicate mean values ( n = 3, Mann-Whitney test, ≥203 fibers were analyzed per condition). ( C ) Effect of expressing WT, K131A, and K131R RAD51C mutants on replication restart following the treatment of U2OS cells with 2 mM HU for the indicated time. Top: Experimental schematic; and bottom: quantification of stalled forks. Data show percentage of stalled forks as means ± SD ( n = 3, two-tailed unpaired t test). ( D and E ) Representative immunofluorescence images (D) and quantification (E) of S9.6 staining after transfection of cells with indicated shRNA and RAD51C constructs. Relative nuclear S9.6 intensity data are plotted as a box plot ( n = 3, two-tailed, unpaired t test with Welch’s correction on means of biological replicates, ≥278 cells were analyzed per condition). Scale bar, 5 μm. P values are indicated. EV, empty vector. h, hours.

Article Snippet: Human cell lines U2OS (American Type Culture Collection; HTB-96; RRID: CVCL_0042), HeLa Kyoto (Sachin Kotak laboratory; RRID: CVCL_1922), and hTERT RPE-1 (Sachin Kotak laboratory) were grown in Dulbecco’s modified Eagle medium supplemented with 10% fetal bovine serum, 1% penicillin/streptomycin (Sigma-Aldrich), and 1% Glutamax (Gibco) under standard conditions (5% CO 2 , humidified atmosphere) at 37°C.

Techniques: Expressing, MANN-WHITNEY, Two Tailed Test, Immunofluorescence, Staining, Transfection, shRNA, Construct, Plasmid Preparation