human crc cell lines ht29  (ATCC)

Journal: Cell Death & Disease

Article Title: Targeting the Met-RIPK1 signaling axis to enforce apoptosis and necroptosis in colorectal cancer

doi: 10.1038/s41419-025-08054-5

Figure Lengend Snippet: A HT29 cells treated with indicated doses of Cabozantinib for 48 h. Remaining fraction of adherent cells quantified via crystal violet. Mean of three independent experiments ± standard deviation (SD) ( n = 3). p < 0.05 (*), p > 0.05 (no significance, n.s.) (Student’s t test). B HT29 cells pre-treated with Cabozantinib (5 μM for two hours), followed by indicated doses of BV6 for 48 h. Fractional survival quantified via crystal violet. Mean of three independent experiments ± SD ( n = 3). p < 0.05 (*), p < 0.01 (**), or p < 0.001 (***) (Student’s t test). C Representative microscopy images illustrating morphological signs of cell death. D Western blot for cleaved Caspase-3 in HT29 cells treated with Cabozantinib and BV6 in the presence or absence of pan-caspase inhibitor ZVAD (25 μM, representative data of three independent experiments). E Fractional cellular survival after 48 h of treatment with BV6 (0.6 μM), Cabozantinib, or ZVAD. Mean of three independent experiments ± SD ( n = 3). p < 0.05 (*) (Student’s t test). F Western blot for pMLKL (representative data of three independent experiments).

Article Snippet: The human CRC cell line HT29 (ATCC) was maintained in RPMI-1640 medium (Gibco, 21875-034) containing 10% FBS (Sigma-Aldrich, F7524) and 1% penicillin/streptomycin (Sigma-Aldrich, P0781) at 5% CO 2 and 37 °C.

Techniques: Standard Deviation, Microscopy, Western Blot

Journal: Cell Death & Disease

Article Title: Targeting the Met-RIPK1 signaling axis to enforce apoptosis and necroptosis in colorectal cancer

doi: 10.1038/s41419-025-08054-5

Figure Lengend Snippet: A HT29 cells treated with Cabozantinib (5 μM) and BV6 (0.6 μM) in the presence or absence of RIPK1 kinase inhibitor Necrostatin-1 (10 μM). Fractional survival after 48 h quantified via crystal violet. Mean of three independent experiments ± standard deviation (SD) ( n = 3). p < 0.01 (**) (Student’s t test). B Fractional survival relative to control cells shown in Fig. 2A after 48 h of indicated treatments, here in the presence of ZVAD (25 μM). Mean of three independent experiments ± SD ( n = 3). p < 0.01 (**), p < 0.001 (***) (Student’s t test). C , D Western blot for PARP and Caspase-3 in HT29 treated with indicated substances for 30 h. Relative quantification of cleaved protein normalized to proform. Mean of three independent experiments ± SD ( n = 3). p < 0.05 (*), p < 0.01 (**), (Student’s t test). E , F Western blot for pRIPK3 and pMLKL in HT29. Relative quantification of phosphorylated normalized to total protein. Mean of three independent experiments ± SD ( n = 3). p > 0.05 (no significance, n.s.), p < 0.01 (**), (Student’s t test). G , H Fractional survival in HT29 wild-type (CRISPR control) and CRISPR/Cas9-knockout (KO) cells after 48 h of indicated treatments. Mean of three independent experiments ± SD ( n = 3). p < 0.05 (*), p < 0.01 (**), p < 0.001 (***) (Student’s t test).

Article Snippet: The human CRC cell line HT29 (ATCC) was maintained in RPMI-1640 medium (Gibco, 21875-034) containing 10% FBS (Sigma-Aldrich, F7524) and 1% penicillin/streptomycin (Sigma-Aldrich, P0781) at 5% CO 2 and 37 °C.

Techniques: Standard Deviation, Control, Western Blot, Quantitative Proteomics, CRISPR, Knock-Out

Journal: Cell Death & Disease

Article Title: Targeting the Met-RIPK1 signaling axis to enforce apoptosis and necroptosis in colorectal cancer

doi: 10.1038/s41419-025-08054-5

Figure Lengend Snippet: A , B Fractional survival in HT29 wild-type (CRISPR control) and CRISPR/Cas9-knockout (TNFR KO) cells after 48 h of indicated treatments (BV6: 0.6 μM, Cabozantinib: 5 μM, ZVAD: 25 μM) quantified via crystal violet. Mean of three independent experiments ± standard deviation (SD) ( n = 3). p < 0.05 (*), p < 0.01 (**) (Student’s t test). C Western blot for pMet and Met in HT29 after indicated treatments. Mean of three independent experiments ± SD ( n = 3). p < 0.01 (**), p < 0.001 (***) (Student’s t test). D–F Relative mRNA expression in HT29 after indicated treatments measured via qRT-PCR. Mean of three independent experiments ± SD ( n = 3). p < 0.05 (*), p < 0.01 (**), p < 0.001 (***) (Student’s t test). G Western blot for RIPK1 in HT29 treated with Cabozantinib for 24 h (representative data of three independent experiments). H Relative quantification of RIPK1 protein in Cabozantinib-treated cells in Fig. 3G. Mean of three independent experiments ± SD ( n = 3). p < 0.05 (*) (Student’s t test). I–K : Relative mRNA expression measured via qRT-PCR. Mean of three independent experiments ± SD ( n = 3). p < 0.05 (*), p < 0.01 (**), p < 0.001 (***) (Student’s t test).

Article Snippet: The human CRC cell line HT29 (ATCC) was maintained in RPMI-1640 medium (Gibco, 21875-034) containing 10% FBS (Sigma-Aldrich, F7524) and 1% penicillin/streptomycin (Sigma-Aldrich, P0781) at 5% CO 2 and 37 °C.

Techniques: CRISPR, Control, Knock-Out, Standard Deviation, Western Blot, Expressing, Quantitative RT-PCR, Quantitative Proteomics

Journal: Cell Death & Disease

Article Title: Targeting the Met-RIPK1 signaling axis to enforce apoptosis and necroptosis in colorectal cancer

doi: 10.1038/s41419-025-08054-5

Figure Lengend Snippet: A HT29 cells were subcutaneously injected into left and right flank of CD1 nude mice. When tumors were palpable (day 0), mice were divided into eight groups with five animals per group to apply apoptotic (left) or necroptotic treatments (right). Every 3 or 4 days, mice received drugs and tumor volumes were measured. Data represent average tumor sizes per group ( n = 10). B Tumor volumes on day 15 for apoptotic (left) and necroptotic treatment groups (right). Each circle represents one individual tumor. Bars are group average, dashed line is average of control group ( n = 10). p < 0.05 (*), p < 0.01 (**), p < 0.001 (***) (one-way ANOVA with Bonferroni’s correction). C Scatter plot to illustrate inverse correlation between tumor sizes and the percentage of tumor area with high RIPK1 expression (see Supplementary Fig. , and Methods section). Each circle represents one individual tumor (total of n = 24, including n = 3 per treatment group) (r, Spearman correlation analysis). D Immunohistochemical stainings of cleaved Caspase-3, pRIPK3 and pMLKL, or Van Gieson’s Elastica (EVG) staining to quantify fibrotic tumor area in apoptotic (left, n = 4) or necroptotic treatment groups (right, n = 3). Quantification depicts percentage of positively stained tumor area; each circle represents one individual tumor, while bars represent mean ± standard error of the mean (SEM). p > 0.05 (no statistical significance, n.s.), p < 0.05 (*), p < 0.01 (**) (Student’s t test). E HMGB1 released by necroptotic HT29 cells after 24 h of treatment. Mean of three independent experiments ± standard deviation (SD) ( n = 3). p < 0.001 (***) (Student’s t test). F Time course of ATP release by necroptotic HT29 cells. Cycloheximide/TNF was included as an inducer of non-inflammatory cell death. Mean of three independent experiments ± SD ( n = 3).

Article Snippet: The human CRC cell line HT29 (ATCC) was maintained in RPMI-1640 medium (Gibco, 21875-034) containing 10% FBS (Sigma-Aldrich, F7524) and 1% penicillin/streptomycin (Sigma-Aldrich, P0781) at 5% CO 2 and 37 °C.

Techniques: Injection, Control, Expressing, Immunohistochemical staining, Staining, Standard Deviation

Journal: Cell Death & Disease

Article Title: Targeting the Met-RIPK1 signaling axis to enforce apoptosis and necroptosis in colorectal cancer

doi: 10.1038/s41419-025-08054-5

Figure Lengend Snippet: A Patient-derived HROC cell lines treated with indicated doses of BV6 for 48 h. Fractional survival quantified via crystal violet assay. Mean of three independent experiments ± standard deviation ( n = 3) (SD). p < 0.05 (*), p < 0.01 (**), or p < 0.001 (***) (Student’s t test). B Western blot for PARP and Caspase-3 in HROC cells treated with BV6 (HROC24, 87: 0.6 μM; HROC46, 69: 5 μM) and ZVAD (25 μM) for 30 h. C Relative mRNA expression measured via qRT-PCR normalized to HT29 as a control. Mean of three independent experiments ± SD ( n = 3). D Fold induction of TNF mRNA expression by BV6 as measured via qRT-PCR. Mean of three independent experiments ± SD ( n = 3). E HROC cell lines treated with BV6 (HROC24, 87: 0.6 μM; HROC39, 40, 46, 69: 5 μM), ZVAD (25 μM) and Necrostatin-1 (10 μM) for 48 h. Fractional survival quantified via crystal violet assay. Mean of three independent experiments ± SD ( n = 3). p < 0.05 (*), p < 0.001 (**) or p < 0.001 (***). (Student’s t test). F Western blot for pMLKL in HROC cells treated as indicated in Fig. 5B. G Relative expression of RIPK3 mRNA measured via qRT-PCR and normalized to HT29. Mean of three independent experiments ± SD ( n = 3). H Cells treated with BV6 and ZVAD as specified in Fig. 5E. TNF secretion quantified in cell culture supernatants via ELISA, and normalized to HROC46. Means of three independent experiments ± SD ( n = 3). I Cells treated as specified in Fig. 5E, in the presence or absence of TNF (25 ng/ml). Fractional survival quantified via crystal violet assay. Mean of three independent experiments ± SD ( n = 3). p < 0.05 (*), p < 0.01 (**) or p < 0.001 (***) (Student’s t test).

Article Snippet: The human CRC cell line HT29 (ATCC) was maintained in RPMI-1640 medium (Gibco, 21875-034) containing 10% FBS (Sigma-Aldrich, F7524) and 1% penicillin/streptomycin (Sigma-Aldrich, P0781) at 5% CO 2 and 37 °C.

Techniques: Derivative Assay, Crystal Violet Assay, Standard Deviation, Western Blot, Expressing, Quantitative RT-PCR, Control, Cell Culture, Enzyme-linked Immunosorbent Assay