panc 1 cell lines  (ATCC)

Journal: bioRxiv

Article Title: Reactivation of DRP1 plays a functional role in resistance to MEK inhibition in pancreatic cancer cells

doi: 10.64898/2026.05.20.726663

Figure Lengend Snippet: a. Western blot of DRP1 from trametinib-resistant 188, 395, PANC-1 and CFPAC-1 cells engineered to stably express a doxycycline-inducible mouse DRP1 and subsequently engineered to express CAS9 and CRISPR guides targeting either a control sequence or endogenous human DRP1. Cells were cultured in the presence or absence of doxycycline for 7 days prior to analysis. b. Inducible DRP1 knockout, trametinib-resistant 188, 395, PANC-1 and CFPAC-1 cells described in (a) were cultured in the absence of doxycycline for 7 days then treated with DMSO or the indicated concentrations of trametinib for 6 days (PANC-1R) or 9 days (188R, 395R, and CFPAC-1R). Viable cells were quantified by CTG assay. N=6 wells per condition. Mean±SD. Three technical replicates with two independent experiments c. Trametinib-resistant 188, 395 and PANC-1 cells were treated with DMSO or a range of abemaciclib doses for 5 days. Viable cells were quantified by CTG assay. N=4 wells per condition. Three independent experiments. Mean±SD. d. Trametinib-resistant 188, 395 and PANC-1 cells were treated with DMSO or a range of trametinib doses either alone, or in combination with 1 μM abemaciclib for 5 days. Viable cells were quantified by CTG assay. N=4 wells per condition. Three independent experiments. Mean±SD. e. 188S and 188RsgCTR cells were injected into the pancreata of nude mice (10 6 cells per mouse) and mice were treated with trametinib (0.3 mg/kg) orally once a day for 4 weeks at which point tumors were harvested and weighed. Unpaired t test, Mean±SD f. Tumors harvested from mice described in (e) were lysed in RIPA and the levels of P-DRP1 (S616) and total DRP1 were analyzed by Western blot. g. 188R sgCTR and 188R sgDRP1 cells were cultured in the absence of doxycycline for 7 days to eliminate expression of mDRP1 and then 10 6 cells were injected cells into the pancreata of nude mice. Mice were treated with either vehicle or trametinib (0.3mg/kg) orally once a day for six weeks and tumors were harvested and weighed. ANOVA. Mean±SD

Article Snippet: Human pancreatic patient-derived xenograft cell lines 188 and 395 [ , ] and established patient-derived pancreatic cancer cell lines PANC-1 and CFPAC-1 (ATCC, VA, USA) were cultured in RPMI 1640 (Gibco #11875093, USA) supplemented with 10% FBS (Gibco #16000-044) and 1% penicillin/streptomycin (Gibco #15140-122).

Techniques: Western Blot, Stable Transfection, CRISPR, Control, Sequencing, Cell Culture, Knock-Out, CTG Assay, Injection, Expressing

Journal: bioRxiv

Article Title: Reactivation of DRP1 plays a functional role in resistance to MEK inhibition in pancreatic cancer cells

doi: 10.64898/2026.05.20.726663

Figure Lengend Snippet: a. Trametinib-sensitive and resistant patient-derived pancreatic cancer cell lines 188, 395, PANC-1 and CFPAC-1 were treated with DMSO or the indicated concentrations of trametinib for 5 days. Viable cells were quantified by CTG assay. N=6 wells per condition. Three technical replicates of each two independent experiments. b. Western blot analysis of P-DRP1 (S616) and total DRP1 from the indicated cells treated DMSO or trametinib (200 nM) for 24hrs. An equal fraction of cells (100,000) was loaded per lane. N=3 to 4 independent experiments. c. Quantification of the blots from b. Data was normalized to corresponding sensitive cells DMSO condition. Mean ± SD. Two-way ANOVA. d. Western blot analysis of the indicated proteins and post-translational modifications from the indicated cells treated DMSO or trametinib (200nM) for 24hrs. An equal fraction of cells (100,000) was loaded per lane. Representative blots from N=3 independent experiments.

Article Snippet: Human pancreatic patient-derived xenograft cell lines 188 and 395 [ , ] and established patient-derived pancreatic cancer cell lines PANC-1 and CFPAC-1 (ATCC, VA, USA) were cultured in RPMI 1640 (Gibco #11875093, USA) supplemented with 10% FBS (Gibco #16000-044) and 1% penicillin/streptomycin (Gibco #15140-122).

Techniques: Derivative Assay, CTG Assay, Western Blot

Journal: bioRxiv

Article Title: Reactivation of DRP1 plays a functional role in resistance to MEK inhibition in pancreatic cancer cells

doi: 10.64898/2026.05.20.726663

Figure Lengend Snippet: a. PCA scores across 24 conditions, including 2 cell lines (188 and PANC-1) × 2 sensitivity conditions (S: sensitive; R: resistant) × 2 treatment conditions (48 h treatment with trametinib at 200 nM, or DMSO), each tested in 3 replicates. Arrows highlight significant trametinib-induced changes in mitochondrial features. b. PC1 loadings of non-redundant mitochondrial features with the strongest influences on PC1. c. Representative graphs of the differentially regulated features following trametinib treatment. The single-cell data for each mitochondrial feature were averaged to obtain a value for each experimental condition. P values represent two-way ANOVA analysis of DMSO vs Trametinib for each cell line. d. Description of features highlighted in (c)

Article Snippet: Human pancreatic patient-derived xenograft cell lines 188 and 395 [ , ] and established patient-derived pancreatic cancer cell lines PANC-1 and CFPAC-1 (ATCC, VA, USA) were cultured in RPMI 1640 (Gibco #11875093, USA) supplemented with 10% FBS (Gibco #16000-044) and 1% penicillin/streptomycin (Gibco #15140-122).

Techniques: Single Cell

Journal: bioRxiv

Article Title: Reactivation of DRP1 plays a functional role in resistance to MEK inhibition in pancreatic cancer cells

doi: 10.64898/2026.05.20.726663

Figure Lengend Snippet: a. Western blot analysis of c-Myc in 188, 395 and PANC-1 sensitive and resistant cells. An equal fraction of cells (100,000) was loaded per lane. N=3 independent experiments. b. Western blot analysis of the indicated proteins and post-translational modifications from trametinib-resistant 188, 395 and PANC-1 cells following 24hr treatment with either DMSO, trametinib (200 nM), 10058-F4 (50 μM), or a combination of trametinib (200 nM) and 10058-F4 (50 μM). 20 μg protein was loaded per lane. N=3 independent experiments. c. Western blot analysis of 188R cells expressing doxycycline inducible shRNA targeting a control sequence or c-Myc treated for 48 hrs with doxycycline to induce knockdown and trametinib (200 nM). N=3 independent experiments. d. Quantification of the blots from c. Welch’s t test. e. Western blot analysis of PANC-1R cells expressing shRNA targeting a control sequence or c-Myc were treated with trametinib (200 nM) for 24hrs.

Article Snippet: Human pancreatic patient-derived xenograft cell lines 188 and 395 [ , ] and established patient-derived pancreatic cancer cell lines PANC-1 and CFPAC-1 (ATCC, VA, USA) were cultured in RPMI 1640 (Gibco #11875093, USA) supplemented with 10% FBS (Gibco #16000-044) and 1% penicillin/streptomycin (Gibco #15140-122).

Techniques: Western Blot, Expressing, shRNA, Control, Sequencing, Knockdown

Journal: bioRxiv

Article Title: Reactivation of DRP1 plays a functional role in resistance to MEK inhibition in pancreatic cancer cells

doi: 10.64898/2026.05.20.726663

Figure Lengend Snippet: c-Myc and CDK4/6 contribute to the DRP1 S616 phosphorylation in trametinib-sensitive pancreatic cancer cells. Western blot analysis of the indicated proteins and post-translational modifications from trametinib-resistant 188, 395 and PANC-1 cells following 24hr treatment with: (a) DMSO, trametinib (200 nM), 10058-F4 (50 μM), or a combination of trametinib (200 nM) and 10058-F4 (50 μM); or (b) DMSO, trametinib (200 nM), abemaciclib (10 μM) or combination of trametinib (200 nM) and abemaciclib (10 μM). 20 μg protein was loaded per lane. N=2 independent experiments.

Article Snippet: Human pancreatic patient-derived xenograft cell lines 188 and 395 [ , ] and established patient-derived pancreatic cancer cell lines PANC-1 and CFPAC-1 (ATCC, VA, USA) were cultured in RPMI 1640 (Gibco #11875093, USA) supplemented with 10% FBS (Gibco #16000-044) and 1% penicillin/streptomycin (Gibco #15140-122).

Techniques: Phospho-proteomics, Western Blot

Journal: bioRxiv

Article Title: Reactivation of DRP1 plays a functional role in resistance to MEK inhibition in pancreatic cancer cells

doi: 10.64898/2026.05.20.726663

Figure Lengend Snippet: a. Western blot analysis of the indicated proteins and post-translational modifications from trametinib-resistant 188, 395 and PANC-1 cells following 24hr treatment with either DMSO, trametinib (200 nM), abemaciclib (10 μM) or combination of trametinib (200 nM) and abemaciclib (10 μM). 20 μg protein was loaded per lane. N=3 independent experiments. b. Western blot analysis of P-DRP1 (S616), CDK4 and CDK6 from trametinib-resistant 188 cells following doxycycline-induced expression of shRNAs targeting a control sequence, CDK4 or CDK6. 20 μg protein was loaded per lane. N=3 independent experiments. c. Quantification of the P-DRP1 levels from the experiment described in d. For each shRNA, P-DRP1 levels in doxycycline-treated cells were normalized to the corresponding untreated control. N=3 independent experiments. Welch’s t test was performed. d. In vitro kinase assay reaction including recombinant GST-DRP1 (518-736) (200 ng) and recombinant His-CDK6/GST-CyclinD1 (25 ng or 50 ng) in presence of ATP. GST-DRP1 (518-736) (SA mutant) was used as a negative control. Reactions were resolved by SDS–PAGE and subjected to either immunoblotting for P-DRP1 S616 and CDK6 or Coomassie staining to visualize recombinant DRP1 protein. N=3 independent experiments. e. Western blot analysis of P-DRP1 (S616) and CDK2 from trametinib-resistant 188 cells following doxycycline-induced expression of shRNA targeting a control sequence or CDK2. N=3 independent experiments. Welch’s t test was performed.

Article Snippet: Human pancreatic patient-derived xenograft cell lines 188 and 395 [ , ] and established patient-derived pancreatic cancer cell lines PANC-1 and CFPAC-1 (ATCC, VA, USA) were cultured in RPMI 1640 (Gibco #11875093, USA) supplemented with 10% FBS (Gibco #16000-044) and 1% penicillin/streptomycin (Gibco #15140-122).

Techniques: Western Blot, Expressing, Control, Sequencing, shRNA, In Vitro, Kinase Assay, Recombinant, Mutagenesis, Negative Control, SDS Page, Staining