human non small cell lung cancer cell line  (ATCC)

Journal: Scientific Reports

Article Title: PISTOL MRI allows noninvasive assessment of changes in tumor oxygenation post hypoxia targeted therapy

doi: 10.1038/s41598-025-32618-2

Figure Lengend Snippet: Overlay of the reference T 2 -weighted tumor image and the respective pO 2 map from day 0–15 for a representative tumor from the control ( a - d ) and TPZ ( e - h )) treated NCI-H1975 cohorts, respectively. The region of interest (shown as red boundary) represents the segmentation that was conducted to delineate the tumor from surrounding tissue and also to segment the tumor into center and periphery. Numbers represent mean pO 2 values ± standard deviation in torr.

Article Snippet: Human NCI-H1975 non-small cell lung cancer and A431 epidermoid carcinomas cells (ATCC Inc.) were cultured in Roswell Park Memorial Institute (RPMI 1640) and Dulbecco’s Modified Eagle’s Medium (DMEM) respectively, supplemented with 1% penicillin/streptomycin and 10% fetal bovine serum (Life Technologies, Carlsbad, CA, USA), in a 5% CO 2 -containing humidified atmosphere at 37 ° C. Immunocompromised male nu\nu mice (5–11/per cohort, Charles River Laboratories, USA) were implanted subcutaneously with 2 × 10 6 cells/40 μl of the respective cell line (A431 or NCI-H1975) in the right thigh.

Techniques: Control, Standard Deviation

Journal: Nature Communications

Article Title: AAK1 activation-mediated iron trafficking drives ferroptotic cell death

doi: 10.1038/s41467-025-67523-9

Figure Lengend Snippet: A Protein levels associated with iron metabolism in various cancer cell lines treated with erastin for 12 h. A549, 8 μM; CAL51, 30 μM; HN6, 30 μM; HT1080, 4 μM; MDA-MB-231, 8 μM; MCF7, 30 μM. B , C Total ( B ) and divalent ( C ) iron levels of various cancer cell lines treated with erastin for 12 h. A549, 4 μM; CAL51, 20 μM; HN6, 20 μM; HT1080, 1 μM; MDA-MB-231, 2 μM; MCF7, 20 μM . D Schematic of the endocytosis assays. TFR1 in the cell surface were labelled with TFR1-specific antibodies at 4 °C followed by transferring cells to 37 °C to allow TFR1 internalization. The internalization levels of TFR1 will be established by antibody-labelled TFR1 from the cell surface evaluated by flow cytometry. This figure was created in BioRender. Lichao, L. (2025) https://BioRender.com/p1ntlvc . E Endocytosis assays of TFR1 in various cancer cell lines treated with erastin for 12 h. A549, 4 μM; CAL51, 20 μM; HN6, 20 μM; HT1080, 1 μM; MDA-MB-231, 2 μM; MCF7, 20 μM. F PKCβ was knocked out using single guide RNAs (sgRNAs) in MDA-MB-231 (left) and HT1080 (right) cells. G Total (left) and divalent (right) iron levels in the indicated MDA-MB-231 cells treated with erastin for 12 h. H , I Endocytosis assays of TFR1 in the indicated MDA-MB-231 ( H ) and HT1080 ( I ) cells treated with 2 μM or 1 μM erastin for 12 h, respectively. The cells with blue highlight were kept in 4 °C for dormancy. The cells with red highlight were transferred to 37 °C for internalization. The cells with grey highlight were labelled with non-specific antibody as isotype control. J , K Plasmids of PKCβⅠ or PKCβⅡ were transfected into PKCβ -knockout MDA-MB-231 ( J ) and HT1080 ( K ) cells, respectively. Endocytosis assays of TFR1 were performed in these cells treated with 2 μM or 1 μM erastin for 12 h, respectively. L Total (left) and divalent (right) iron levels in the indicated MDA-MB-231 cells treated with erastin for 12 h. A , F , J , K Data are representative of n = 3 biologically independent experiments. B , C , E , J , K Data are presented as means ± SD, n = 3 biologically independent experiments, unpaired two-tailed Student’s t test. G–I , L Data are presented as means ± SD, n = 3 biologically independent experiments, one-way ANOVA test.

Article Snippet: Human breast cancer cell MDA-MB-231 and MCF7, human fibrosarcoma cell HT1080, human Non-Small Cell Lung Cancer cell A549 were obtained from American Type Culture Collection.

Techniques: Transferring, Flow Cytometry, Control, Transfection, Knock-Out, Two Tailed Test

Journal: Scientific Reports

Article Title: Statins attenuate PD-L1 sorting to small extracellular vesicles dependent on ubiquitin-like 3 modification

doi: 10.1038/s41598-025-27789-x

Figure Lengend Snippet: UBL3 regulates PD-L1 levels in sEVs. ( a ) Immunoblot (IB) analysis. The cell lysates and sEVs from the conditioned medium of H1299 cells transfected with 3xFlag-UBL3 vectors were blotted with various antibodies. ( b ) IB analysis. The cell lysates and sEVs from the stable UBL3 knockdown H1299 cell lines were blotted with various antibodies. CD9 and CD63 were used as sEV markers, as they are tetraspanin proteins abundantly expressed on the sEV surface . Relative intensity of PD-L1 in sEVs was calculated as PD-L1/CD63 and presented as fold change, normalized to the mean value of the respective control (mock for a; shNega for b). Data are presented as mean ± s.e.m., with dots representing individual experiments. Two-tailed unpaired t-test. *** P < 0.005.

Article Snippet: Human non-small cell lung cancer H1299 cell (CRL-5803, ATCC), human melanoma WM9 cells (WM9-01-0001, Rockland) and human breast cancer cell MDA-MB-231(MDA-MB-231-luc-D3H2LN, Xenogen) were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium (11875-093, Gibco) with 10% heat-inactivated fetal bovine serum (FBS; SH30910.03, Cytiva).

Techniques: Western Blot, Transfection, Knockdown, Control, Two Tailed Test

Journal: Scientific Reports

Article Title: Statins attenuate PD-L1 sorting to small extracellular vesicles dependent on ubiquitin-like 3 modification

doi: 10.1038/s41598-025-27789-x

Figure Lengend Snippet: Inhibition of UBL3 modification and PD-L1 sorting by statins. ( a and b ), Effect of pitavastatin treatment on UBL3 modification in WM9 ( a ) and H1299 ( b ) cells. Five hours after transfection, 10 µM pitavastatin was added. ( c and d ), IB analysis of the cell lysates and sEVs from the conditioned medium of H1299 cells transfected with 3xFlag-UBL3 vectors ( c ) or untransfected ( d ) were blotted with various antibodies. Five hours after gene transfection, 0.2 µM pitavastatin was added. Pit, Pitavastatin. Right panels, the relative intensity of PD-L1 in sEVs (c: PD-L1/CD63 [upper] normalized to the mean of mock − untreated control, and UBL3/CD63 [lower] normalized to the mean of Flag-UBL3 − untreated control; d: PD-L1/CD63 [upper] and UBL3/CD63 [lower], both normalized to the mean of untreated control). Data are presented as mean ± s.e.m., with dots representing individual experiments. As inhibition of UBL3 modification was observed even at low concentrations of pitavastatin (Supplementary Fig. S4b), a low concentration of pitavastatin was used in sEV purification experiments. IB analysis using the indicated antibodies. Flag, Flag-tag; GAPDH, internal control; CD63 and CD9, sEVs markers; PD-L1, endogenous PD-L1; UBL3, endogenous UBL3. βME−, without 2-mercaptoethanol. βME+, with 2-mercaptoethanol. ( c ), one-way ANOVA with Tukey’s multiple comparisons test. ( d ), Two-tailed unpaired t-test. * P < 0.05, ** P < 0.01.

Article Snippet: Human non-small cell lung cancer H1299 cell (CRL-5803, ATCC), human melanoma WM9 cells (WM9-01-0001, Rockland) and human breast cancer cell MDA-MB-231(MDA-MB-231-luc-D3H2LN, Xenogen) were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium (11875-093, Gibco) with 10% heat-inactivated fetal bovine serum (FBS; SH30910.03, Cytiva).

Techniques: Inhibition, Modification, Transfection, Control, Concentration Assay, Purification, FLAG-tag, Two Tailed Test