nheks  (ATCC)

Journal: Communications Biology

Article Title: The MYB-related transcription factor MYPOP acts as a selective regulator of cancer cell growth

doi: 10.1038/s42003-026-10272-2

Figure Lengend Snippet: a MYPOP expression in normal (N) and tumor (T) cells detected by Western blotting showing endogenous protein levels of MYPOP in normal human epidermal keratinocytes (NHEK) and cervical cancer cells (HeLa). In addition, the expression of GFP and MYPOP in HeLa cells after transfection with pcDNA3.1-MYPOP (MYPOP), pEGFP-C3-MYPOP (GFP-MYPOP) or corresponding controls (pcDNA3.1, Control and pEGFP-C3, GFP-Control) is shown. MYPOP and GFP were stained using anti-MYPOP pAb and anti-GFP mAb. GAPDH staining was used as a loading control. b – i GFP-Control or GFP-MYPOP expressing HeLa cells. b , c Lower ( b ) and higher ( c ) magnification for representative fluorescence microscopy images of GFP-Control or GFP-MYPOP (green) expressing cells 24 h post transfection (p.t.). Cell nuclei were stained using Hoechst 33342 (blue). d , e Representative fluorescence microscopy images treated as in ( c ) showing cytoplasmic, nucleocytoplasmic ( d ) and nuclear ( e ) localization as well as co-localization of GFP-MYPOP (green) and DNA (blue). f Co-localization analysis between GFP or GFP-MYPOP and DNA (Hoechst) using Pearson correlation coefficient (PCC). At least 10 GFP-positive cells were analyzed for each treatment and biological replicate ( n = 3). Values are shown as mean + SD. Statistical significance was determined with p = 0.0001 comparing GFP-MYPOP and GFP-Control. g Representative fluorescence microscopy images of GFP-MYPOP (green) expressing cells showing shrunken or fragmented nuclei at 24 h p.t. Cell nuclei (blue) as above. h , i Quantification of fragmented and shrunken nuclei at 24 h p.t ( h ) and 48 h p.t ( i ). At least 100 GFP-positive cells were analyzed for each treatment, time point and biological replicate, respectively. Values ( n = 3) are shown as mean + SD. Statistical significance was determined with p (24 h p.t.) = 0.0036 and p (48 h p.t.) = 0.0428 comparing GFP-MYPOP and GFP-Control.

Article Snippet: Normal Human Epidermal Keratinocytes (NHEK) were purchased from PromoCell, Germany and were cultivated according to the manufacturer’s instructions.

Techniques: Expressing, Western Blot, Transfection, Control, Staining, Fluorescence, Microscopy

Journal: Communications Biology

Article Title: The MYB-related transcription factor MYPOP acts as a selective regulator of cancer cell growth

doi: 10.1038/s42003-026-10272-2

Figure Lengend Snippet: a Endogenous protein levels of MYPOP in untreated cancer cell lines. Normal skin cells (NHEK) and normal lung cells (181576 N, 181652 N) served as controls. GAPDH or β-actin staining was used as loading control. b Cancer cells were transfected with either a MYPOP expression plasmid or a control plasmid, selected for 6–12 days with G418, fixed, and stained with crystal violet. The cell-covered area was quantified (relative area). Values are presented as mean + SD. The mean for control-transfected cells was set to 100% (dotted line). Statistical significance was determined by comparing control and MYPOP expressing cells with p = 0.0022 for Huh7 ( n = 3), p = 0.0028 for HEK293 ( n = 5), p = 0.0064 for MCF7 ( n = 4), p = 0.0294 for HCT116 ( n = 3), p = 0.0106 for HeLa ( n = 3), p = 0.0003 for A549 ( n = 3), and p = 0.0026 for 2106 T ( n = 4).

Article Snippet: Normal Human Epidermal Keratinocytes (NHEK) were purchased from PromoCell, Germany and were cultivated according to the manufacturer’s instructions.

Techniques: Staining, Control, Transfection, Expressing, Plasmid Preparation

Journal: Communications Biology

Article Title: The MYB-related transcription factor MYPOP acts as a selective regulator of cancer cell growth

doi: 10.1038/s42003-026-10272-2

Figure Lengend Snippet: a Protein expression of untreated, control mRNA and MYPOP mRNA-transfected HeLa cells and NHEK cells, as indicated at different time points post transfection (6 h–54 h) was analyzed by western blotting using anti-MYPOP and anti-GAPDH antibodies. Under conditions of low protein loading and short exposure times, endogenous MYPOP is not detected in NHEK, as these parameters are optimized to prevent oversaturation of overexpressed MYPOP at 6 h p.t. Detection of endogenous MYPOP increases at later time points, reflecting cell growth and higher total protein content. Lower panel: optical microscope overview images of untreated, control mRNA and MYPOP mRNA-transfected HeLa cells (left panel) and NHEK cells (right panel) at 6 h and 54 h after mRNA transfection. b Growth curves (object counts per image, measurement every 2 h) of untreated, control mRNA and MYPOP mRNA-transfected HeLa, NHEK, CaSki, HaCaT, HCT116, and 2106 T cells at the indicated time points. Statistical significance ( n = 4 for HeLa, n = 3 for all others) was determined between Control and MYPOP cell counts at 70 h or 72 h p.t. as indicated with p = 0.0203 for HeLa, p = 0.0441 for CaSki, p = 0.4720 for HCT116, p = 0.2424 for NHEK, p < 0.0001 for HaCaT, and p = 0.0152 for 2106 T. c Left: MYPOP and GAPDH protein expression in Hela wild-type (WT) and MYPOP knockout (KO) cells. Statistical significance ( n = 3) between WT and KO cells was determined with p = 0.0014 for MYPOP band intensities. Center: optical microscope overview images of HeLa WT and KO cells. Right: growth curve of HeLa WT and KO cells. Statistical significance ( n = 4) was determined with p = 0.6099 at 72 h t.p.

Article Snippet: Normal Human Epidermal Keratinocytes (NHEK) were purchased from PromoCell, Germany and were cultivated according to the manufacturer’s instructions.

Techniques: Expressing, Control, Transfection, Western Blot, Microscopy, Knock-Out

Journal: Communications Biology

Article Title: The MYB-related transcription factor MYPOP acts as a selective regulator of cancer cell growth

doi: 10.1038/s42003-026-10272-2

Figure Lengend Snippet: Volcano plots depicting gene expression changes of control and MYPOP mRNA-transfected HeLa and NHEK cells at 6 h and 24 h p.t. as indicated. The labels display the overlapping DEGs, which were selected from the initial RNA-Seq experiment shown in Fig. , comprising the 30 top DEGs (15 up, 15 downregulated) and the 27 ‘cell cycle’ genes. Significantly downregulated candidates with adjusted p ≤ 0.05 are shown in blue, and upregulated candidates with adjusted p ≤ 0.05 are shown in red.

Article Snippet: Normal Human Epidermal Keratinocytes (NHEK) were purchased from PromoCell, Germany and were cultivated according to the manufacturer’s instructions.

Techniques: Gene Expression, Control, Transfection, RNA Sequencing

Journal: International Journal of Molecular Sciences

Article Title: A Functional Evaluation of Resveratrol–Paclitaxel Combination Reveals Enhanced Apoptotic Responses in HeLa Cells

doi: 10.3390/ijms27104505

Figure Lengend Snippet: Limited selectivity in immortalized normal cells of RES and PAC in HeLa and HaCaT cells. ( A ) IC 50 values for RES in HeLa cervical cancer cells and HaCaT normal keratinocytes at 24 and 48 h. ( B ) IC 50 values for PAC under the same conditions. For both agents, IC 50 values were consistently higher in HaCaT cells compared to HeLa cells at both time points, indicating reduced sensitivity of normal cells. The selectivity index (SI), calculated as IC 50 (HaCaT)/IC 50 (HeLa), exceeded 2 in all conditions (RES: 2.6 and 2.2; PAC: 2.2 and 2.1 at 24 and 48 h, respectively), indicating limited selectivity in immortalized normal cells rather than definitive tumor-specific selectivity. Data are derived from three independent experiments ( n = 3). IC 50 values were calculated by nonlinear regression analysis of dose–response curves. It should be noted that HaCaT cells are immortalized keratinocytes and do not fully represent primary normal cervical epithelial cells.

Article Snippet: Human cervical carcinoma HeLa cells (ATCC ® CCL-2TM) and human immortalized keratinocyte HaCaT cells (Cell Lines Service, CLS, Cat. No. 300493, Eppelheim, Germany) were used in this study.

Techniques: Derivative Assay