drug treatment human colorectal cancer cell lines dld 1  (ATCC)

Journal: Cell death & disease

Article Title: Hypoxia-induced ATF3 escalates breast cancer invasion by increasing collagen deposition via P4HA1.

doi: 10.1038/s41419-025-07461-y

Figure Lengend Snippet: Fig. 5 P4HA1 9a isoform increases the invasiveness of hypoxic breast cancer cells. A Invasion assay and B its quantification in control and P4HA1 9a or 9b isoform overexpressing MCF7 cells under normoxic condition. C Invasion assay and D its quantification in P4HA1 knockdown MCF7 cells rescued by overexpressing P4HA1 9a or 9b isoform as compared to the control P4HA1 knockdown cells as well as the control cells transfected with a non-targeting shRNA in hypoxic conditions. E Immunostaining of COL1A1 in ATF3 knockdown MCF7 cells which is then rescued by P4HA1 9a overexpression (DAPI staining is done to show the nucleus and overexpression of the P4HA1 isoforms is confirmed by GFP, scale bars, 10 µm). Error bars show mean values ± SD (n = 3 (biological replicates), unless otherwise specified) as calculated using two- tailed Student’s t-test, unless otherwise specified, *P ≤0.05, **P ≤0.01, ***P ≤0.001, ns, not significant.

Article Snippet: MATERIALS AND METHODS Cell culture and hypoxia treatment Human breast cancer cell lines MCF7 (ATCC HTB22) [28] and HCC1806 (ATCC CRL2335) [29] obtained from American Type Culture Collection (ATCC) were cultured in DMEM (Invitrogen, 12800017) and RPMI (Invitrogen, 23400021), respectively, HEK293T (ATCC CRL1573) was cultured in DMEM, Primary breast cell lines were cultured in DMEM/ F12(1:1).

Techniques: Invasion Assay, Control, Knockdown, Transfection, shRNA, Immunostaining, Over Expression, Staining, Two Tailed Test