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Cold Spring Harbor Laboratory Meetings pdx1 cre kpc mice kpc mt4

CRISPR-mediated KDM4C deletion attenuates PDAC growth in vitro and in vivo . A, Western blot showing KDM4C depletion in selected AsPC1 and <t>KPC-mT4</t> KDM4C -null clones. B, Colony formation assay (CFA): representative wells comparing AsPC1 cells to KDM4C KO isogenic cells show reduced colony formation ability in the absence of KDM4C. C, Quantification of CFA by ImageJ shows a significant reduction in the number of colonies in KDM4C KO cells compared with KDM4C WT cells; P value was determined by one-way ANOVA test; *, P ≤ 0.05; **, P ≤ 0.01. D, Proliferation assay measured through % confluence over time by the Incucyte Live Imaging system shows decreased proliferation in the absence of KDM4C, P value determined by one-way ANOVA test. E, Kaplan–Meier survival plot showing increased survival of B6 mice orthotopically transplanted with KDM4C KO KPC-mT4 cells compared with those transplanted with KDM4C WT KPC-mT4 ( n = 6 per arm).

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1) Product Images from "The Lysine Demethylase KDM4C Is an Oncogenic Driver and Regulates ERK Activity in KRAS-Mutant Pancreatic Ductal Adenocarcinoma"

Article Title: The Lysine Demethylase KDM4C Is an Oncogenic Driver and Regulates ERK Activity in KRAS-Mutant Pancreatic Ductal Adenocarcinoma

Journal: Cancer Research Communications

doi: 10.1158/2767-9764.CRC-25-0278

CRISPR-mediated KDM4C deletion attenuates PDAC growth in vitro and in vivo . A, Western blot showing KDM4C depletion in selected AsPC1 and KPC-mT4 KDM4C -null clones. B, Colony formation assay (CFA): representative wells comparing AsPC1 cells to KDM4C KO isogenic cells show reduced colony formation ability in the absence of KDM4C. C, Quantification of CFA by ImageJ shows a significant reduction in the number of colonies in KDM4C KO cells compared with KDM4C WT cells; P value was determined by one-way ANOVA test; *, P ≤ 0.05; **, P ≤ 0.01. D, Proliferation assay measured through % confluence over time by the Incucyte Live Imaging system shows decreased proliferation in the absence of KDM4C, P value determined by one-way ANOVA test. E, Kaplan–Meier survival plot showing increased survival of B6 mice orthotopically transplanted with KDM4C KO KPC-mT4 cells compared with those transplanted with KDM4C WT KPC-mT4 ( n = 6 per arm).

Figure Legend Snippet: CRISPR-mediated KDM4C deletion attenuates PDAC growth in vitro and in vivo . A, Western blot showing KDM4C depletion in selected AsPC1 and KPC-mT4 KDM4C -null clones. B, Colony formation assay (CFA): representative wells comparing AsPC1 cells to KDM4C KO isogenic cells show reduced colony formation ability in the absence of KDM4C. C, Quantification of CFA by ImageJ shows a significant reduction in the number of colonies in KDM4C KO cells compared with KDM4C WT cells; P value was determined by one-way ANOVA test; *, P ≤ 0.05; **, P ≤ 0.01. D, Proliferation assay measured through % confluence over time by the Incucyte Live Imaging system shows decreased proliferation in the absence of KDM4C, P value determined by one-way ANOVA test. E, Kaplan–Meier survival plot showing increased survival of B6 mice orthotopically transplanted with KDM4C KO KPC-mT4 cells compared with those transplanted with KDM4C WT KPC-mT4 ( n = 6 per arm).

Techniques Used: CRISPR, In Vitro, In Vivo, Western Blot, Clone Assay, Colony Assay, Proliferation Assay, Imaging

KDM4C expression correlates with ERK activity. A, Top enriched IPA pathways from RNA-seq data from KDM4C KO vs. WT AsPC1 are cell-cycle arrest, inhibition of MEK/ERK, protein synthesis inhibition, and increased oxidative phosphorylation and apoptosis, whereas ATF4, MYC, and ERK1/2 are among the top downregulated networks. B, GSEA plot from RNA-seq data showing downregulated KRAS and MYC signatures in KDM4C -depleted cells. C, RPPA data demonstrate downregulation of the ERK pathway in two different KDM4C -null clones. D, Western blot validating downregulation of ERK phosphorylation in human PDAC cell line AsPC1 and in mouse PDAC cell line (KPC-mT4) KDM4C KO clones. E, Representative micrographs of pERK expression by IHC in PDAC tumors harvested at day 30 from mice injected with PDAC cell line KPC-mT4 demonstrates downregulation of pERK in the KDM4C KO clones compared with untreated KPC-mT4 (Scale bar, 100 μm).

Figure Legend Snippet: KDM4C expression correlates with ERK activity. A, Top enriched IPA pathways from RNA-seq data from KDM4C KO vs. WT AsPC1 are cell-cycle arrest, inhibition of MEK/ERK, protein synthesis inhibition, and increased oxidative phosphorylation and apoptosis, whereas ATF4, MYC, and ERK1/2 are among the top downregulated networks. B, GSEA plot from RNA-seq data showing downregulated KRAS and MYC signatures in KDM4C -depleted cells. C, RPPA data demonstrate downregulation of the ERK pathway in two different KDM4C -null clones. D, Western blot validating downregulation of ERK phosphorylation in human PDAC cell line AsPC1 and in mouse PDAC cell line (KPC-mT4) KDM4C KO clones. E, Representative micrographs of pERK expression by IHC in PDAC tumors harvested at day 30 from mice injected with PDAC cell line KPC-mT4 demonstrates downregulation of pERK in the KDM4C KO clones compared with untreated KPC-mT4 (Scale bar, 100 μm).

Techniques Used: Expressing, Activity Assay, RNA Sequencing, Inhibition, Phospho-proteomics, Clone Assay, Western Blot, Injection

Compensatory upregulation of KDM4A restores cell-intrinsic ERK signaling in KDM4C -null cells but not immune surveillance. A, Western blot panel shows that late-passage KDM4C KO clones of both AsPC1 and KPC-mT4 (KO-late) restored pERK levels. B–D, ERK reactivation rescues the reduction in colony formation and proliferation in both AsPC1 and KPC-mT4 KO cells observed in earlier passages. Bar plots show number of colonies calculated by ImageJ. For each sample, two wells have been analyzed, and the statistical significance is calculated using one-way ANOVA. For both AsPC1 and KPC-mT4 cells, the difference between parental and adapted KDM4C KO clones was not significant. E, Lentiviral shRNA knockdown of KDM4C recapitulates the adaptation to KDM4C depletion in adapted cells. Western blot panel compares early-passage (day 7 after transduction) and late-passage (day 40 after transduction) in AsPC1 cells. Late-passage KDM4C knockdown cells have adapted to the loss of KDM4C and restored ERK activation. F, Western blot validating KDM4A upregulation in adapted KPC-mT4 KDM4C KO clones. G, RNA-seq results from AsPC1 early-passage KDM4C KO cells correlate with the ERK inhibitor (ERKi) transcriptome signature, whereas late-passage adapted cells are inversely correlated. H and I, Bar graphs showing significant reduction in the tumor growth of adapted KPC-mT4 KDM4C KO cells when transplanted into the immunocompetent B6 mice compared with KDM4C WT control ( n = 5), whereas the reverse is observed when the same cells are transplanted into immune-compromised athymic mice ( n = 8), significance determined by unpaired t test; *, P ≤ 0.05.

Figure Legend Snippet: Compensatory upregulation of KDM4A restores cell-intrinsic ERK signaling in KDM4C -null cells but not immune surveillance. A, Western blot panel shows that late-passage KDM4C KO clones of both AsPC1 and KPC-mT4 (KO-late) restored pERK levels. B–D, ERK reactivation rescues the reduction in colony formation and proliferation in both AsPC1 and KPC-mT4 KO cells observed in earlier passages. Bar plots show number of colonies calculated by ImageJ. For each sample, two wells have been analyzed, and the statistical significance is calculated using one-way ANOVA. For both AsPC1 and KPC-mT4 cells, the difference between parental and adapted KDM4C KO clones was not significant. E, Lentiviral shRNA knockdown of KDM4C recapitulates the adaptation to KDM4C depletion in adapted cells. Western blot panel compares early-passage (day 7 after transduction) and late-passage (day 40 after transduction) in AsPC1 cells. Late-passage KDM4C knockdown cells have adapted to the loss of KDM4C and restored ERK activation. F, Western blot validating KDM4A upregulation in adapted KPC-mT4 KDM4C KO clones. G, RNA-seq results from AsPC1 early-passage KDM4C KO cells correlate with the ERK inhibitor (ERKi) transcriptome signature, whereas late-passage adapted cells are inversely correlated. H and I, Bar graphs showing significant reduction in the tumor growth of adapted KPC-mT4 KDM4C KO cells when transplanted into the immunocompetent B6 mice compared with KDM4C WT control ( n = 5), whereas the reverse is observed when the same cells are transplanted into immune-compromised athymic mice ( n = 8), significance determined by unpaired t test; *, P ≤ 0.05.

Techniques Used: Western Blot, Clone Assay, shRNA, Knockdown, Transduction, Activation Assay, RNA Sequencing, Control

Pan-KDM4 Inhibitor TACH107 reduces proliferation and colony formation in PDAC cell lines in vitro and increases survival in vivo . A, MTT assay shows reduced proliferation in PDAC cell lines following TACH107 treatment, significance determined by one-way ANOVA. B, Reduced colony formation ability of human and mouse PDAC cell lines upon treatment with varying doses of TACH107 and ( C ) Quantification of the number of colonies, one-way ANOVA was used to assess significance. D, Survival plot showing B6 mice transplanted with PDAC cell line KPC-mT4 and treated with TACH107 survive longer than those treated with vehicle only ( n = 3). E, Results from CETSA show increased heat stability of KDM4 proteins when treated with TACH107 vs. DMSO. F, Representative IF images showing increased H3K9me3 signal after TACH107 treatment compared with control cells. G, Normalized fluorescence intensity of H3K9me3 signal calculated by the corrected total cell fluorescence method using ImageJ, significance determined via unpaired Student t test. For all plots; * is P ≤ 0.05; ** is P ≤ 0.01; *** is P ≤ 0.001; and **** is P ≤ 0.0001. H, Representative micrographs showing H3K9me3 levels by IHC in PDAC tumors harvested on day 30 from mice injected with PDAC cell line KPC-mT4 demonstrate increased H3K9me3 mark in the TACH107 treated mice compared with vehicle control mice. Scale bar, 100 μm. I, Quantification of the area of positive staining in H using ImageJ color deconvolution plugin for IHC DAB staining, the difference between vehicle-treated and TACH107-treated samples is statistically significant.

Figure Legend Snippet: Pan-KDM4 Inhibitor TACH107 reduces proliferation and colony formation in PDAC cell lines in vitro and increases survival in vivo . A, MTT assay shows reduced proliferation in PDAC cell lines following TACH107 treatment, significance determined by one-way ANOVA. B, Reduced colony formation ability of human and mouse PDAC cell lines upon treatment with varying doses of TACH107 and ( C ) Quantification of the number of colonies, one-way ANOVA was used to assess significance. D, Survival plot showing B6 mice transplanted with PDAC cell line KPC-mT4 and treated with TACH107 survive longer than those treated with vehicle only ( n = 3). E, Results from CETSA show increased heat stability of KDM4 proteins when treated with TACH107 vs. DMSO. F, Representative IF images showing increased H3K9me3 signal after TACH107 treatment compared with control cells. G, Normalized fluorescence intensity of H3K9me3 signal calculated by the corrected total cell fluorescence method using ImageJ, significance determined via unpaired Student t test. For all plots; * is P ≤ 0.05; ** is P ≤ 0.01; *** is P ≤ 0.001; and **** is P ≤ 0.0001. H, Representative micrographs showing H3K9me3 levels by IHC in PDAC tumors harvested on day 30 from mice injected with PDAC cell line KPC-mT4 demonstrate increased H3K9me3 mark in the TACH107 treated mice compared with vehicle control mice. Scale bar, 100 μm. I, Quantification of the area of positive staining in H using ImageJ color deconvolution plugin for IHC DAB staining, the difference between vehicle-treated and TACH107-treated samples is statistically significant.

Techniques Used: In Vitro, In Vivo, MTT Assay, Control, Fluorescence, Injection, Staining

Image Search Results

Journal: Cancer Research Communications

Article Title: The Lysine Demethylase KDM4C Is an Oncogenic Driver and Regulates ERK Activity in KRAS-Mutant Pancreatic Ductal Adenocarcinoma

doi: 10.1158/2767-9764.CRC-25-0278

Figure Lengend Snippet: CRISPR-mediated KDM4C deletion attenuates PDAC growth in vitro and in vivo . A, Western blot showing KDM4C depletion in selected AsPC1 and KPC-mT4 KDM4C -null clones. B, Colony formation assay (CFA): representative wells comparing AsPC1 cells to KDM4C KO isogenic cells show reduced colony formation ability in the absence of KDM4C. C, Quantification of CFA by ImageJ shows a significant reduction in the number of colonies in KDM4C KO cells compared with KDM4C WT cells; P value was determined by one-way ANOVA test; *, P ≤ 0.05; **, P ≤ 0.01. D, Proliferation assay measured through % confluence over time by the Incucyte Live Imaging system shows decreased proliferation in the absence of KDM4C, P value determined by one-way ANOVA test. E, Kaplan–Meier survival plot showing increased survival of B6 mice orthotopically transplanted with KDM4C KO KPC-mT4 cells compared with those transplanted with KDM4C WT KPC-mT4 ( n = 6 per arm).

Article Snippet: Human PDAC cell lines AsPC1 (RRID: CVCL_0152) and Pa03C (RRID: CVCL_E301) and murine PDAC cell line derived from KRAS LSL-G12D/+ , Trp53 LSL-R172H/+ , Pdx1-Cre (KPC) mice KPC mT4 (Dr. David Tuveson, Cold Spring Harbor Laboratories; ref. ) as well as HPNE (RRID: CVCL_C466) and HEK293 cell lines (RRID: CVCL_0045) were used.

Techniques: CRISPR, In Vitro, In Vivo, Western Blot, Clone Assay, Colony Assay, Proliferation Assay, Imaging

Journal: Cancer Research Communications

Article Title: The Lysine Demethylase KDM4C Is an Oncogenic Driver and Regulates ERK Activity in KRAS-Mutant Pancreatic Ductal Adenocarcinoma

doi: 10.1158/2767-9764.CRC-25-0278

Figure Lengend Snippet: KDM4C expression correlates with ERK activity. A, Top enriched IPA pathways from RNA-seq data from KDM4C KO vs. WT AsPC1 are cell-cycle arrest, inhibition of MEK/ERK, protein synthesis inhibition, and increased oxidative phosphorylation and apoptosis, whereas ATF4, MYC, and ERK1/2 are among the top downregulated networks. B, GSEA plot from RNA-seq data showing downregulated KRAS and MYC signatures in KDM4C -depleted cells. C, RPPA data demonstrate downregulation of the ERK pathway in two different KDM4C -null clones. D, Western blot validating downregulation of ERK phosphorylation in human PDAC cell line AsPC1 and in mouse PDAC cell line (KPC-mT4) KDM4C KO clones. E, Representative micrographs of pERK expression by IHC in PDAC tumors harvested at day 30 from mice injected with PDAC cell line KPC-mT4 demonstrates downregulation of pERK in the KDM4C KO clones compared with untreated KPC-mT4 (Scale bar, 100 μm).

Techniques: Expressing, Activity Assay, RNA Sequencing, Inhibition, Phospho-proteomics, Clone Assay, Western Blot, Injection

Journal: Cancer Research Communications

Article Title: The Lysine Demethylase KDM4C Is an Oncogenic Driver and Regulates ERK Activity in KRAS-Mutant Pancreatic Ductal Adenocarcinoma

doi: 10.1158/2767-9764.CRC-25-0278

Figure Lengend Snippet: Compensatory upregulation of KDM4A restores cell-intrinsic ERK signaling in KDM4C -null cells but not immune surveillance. A, Western blot panel shows that late-passage KDM4C KO clones of both AsPC1 and KPC-mT4 (KO-late) restored pERK levels. B–D, ERK reactivation rescues the reduction in colony formation and proliferation in both AsPC1 and KPC-mT4 KO cells observed in earlier passages. Bar plots show number of colonies calculated by ImageJ. For each sample, two wells have been analyzed, and the statistical significance is calculated using one-way ANOVA. For both AsPC1 and KPC-mT4 cells, the difference between parental and adapted KDM4C KO clones was not significant. E, Lentiviral shRNA knockdown of KDM4C recapitulates the adaptation to KDM4C depletion in adapted cells. Western blot panel compares early-passage (day 7 after transduction) and late-passage (day 40 after transduction) in AsPC1 cells. Late-passage KDM4C knockdown cells have adapted to the loss of KDM4C and restored ERK activation. F, Western blot validating KDM4A upregulation in adapted KPC-mT4 KDM4C KO clones. G, RNA-seq results from AsPC1 early-passage KDM4C KO cells correlate with the ERK inhibitor (ERKi) transcriptome signature, whereas late-passage adapted cells are inversely correlated. H and I, Bar graphs showing significant reduction in the tumor growth of adapted KPC-mT4 KDM4C KO cells when transplanted into the immunocompetent B6 mice compared with KDM4C WT control ( n = 5), whereas the reverse is observed when the same cells are transplanted into immune-compromised athymic mice ( n = 8), significance determined by unpaired t test; *, P ≤ 0.05.

Techniques: Western Blot, Clone Assay, shRNA, Knockdown, Transduction, Activation Assay, RNA Sequencing, Control

Journal: Cancer Research Communications

Article Title: The Lysine Demethylase KDM4C Is an Oncogenic Driver and Regulates ERK Activity in KRAS-Mutant Pancreatic Ductal Adenocarcinoma

doi: 10.1158/2767-9764.CRC-25-0278

Figure Lengend Snippet: Pan-KDM4 Inhibitor TACH107 reduces proliferation and colony formation in PDAC cell lines in vitro and increases survival in vivo . A, MTT assay shows reduced proliferation in PDAC cell lines following TACH107 treatment, significance determined by one-way ANOVA. B, Reduced colony formation ability of human and mouse PDAC cell lines upon treatment with varying doses of TACH107 and ( C ) Quantification of the number of colonies, one-way ANOVA was used to assess significance. D, Survival plot showing B6 mice transplanted with PDAC cell line KPC-mT4 and treated with TACH107 survive longer than those treated with vehicle only ( n = 3). E, Results from CETSA show increased heat stability of KDM4 proteins when treated with TACH107 vs. DMSO. F, Representative IF images showing increased H3K9me3 signal after TACH107 treatment compared with control cells. G, Normalized fluorescence intensity of H3K9me3 signal calculated by the corrected total cell fluorescence method using ImageJ, significance determined via unpaired Student t test. For all plots; * is P ≤ 0.05; ** is P ≤ 0.01; *** is P ≤ 0.001; and **** is P ≤ 0.0001. H, Representative micrographs showing H3K9me3 levels by IHC in PDAC tumors harvested on day 30 from mice injected with PDAC cell line KPC-mT4 demonstrate increased H3K9me3 mark in the TACH107 treated mice compared with vehicle control mice. Scale bar, 100 μm. I, Quantification of the area of positive staining in H using ImageJ color deconvolution plugin for IHC DAB staining, the difference between vehicle-treated and TACH107-treated samples is statistically significant.

Techniques: In Vitro, In Vivo, MTT Assay, Control, Fluorescence, Injection, Staining

(A) Histological analysis (H&E and Masson’s trichrome staining). Scale bar, 200 μm. The enlarged boxed regions highlight details. Scale bar, 100 μm. (B) PDAC progression stages ( n = 7–10 mice for each genotype). (C) Immunohistochemistry of pancreas stained for Ki67 and α-SMA. Scale bars, 20 μm for Ki67 and 100 μm α-SMA. (D) Quantification of Ki67-positive nuclei and α-SMA-positive area (mean ± SD, n = 4 mice for each genotype). (E) Representative pancreas images. Scale bar, 1 cm. (F) Pancreas weight quantification (mean ± SD, n = 6 WT, 7 KPC, 9 KPC::RHOA-KO mice). (G) Survival curve of KPC and KPC::RHOA-KO mice ( n = 10 mice for each genotype). (H) H&E-stained pancreas sections of KPC and KPC::RHOA-KO mice at 12 weeks post-tamoxifen injection. Scale bar, 1 cm. Tumor boundaries are outlined; boxed regions are enlarged. (I) Quantification of inverse circularity of tumors (mean ± SD, n = 20 sections from 6 KPC mice and 50 sections from 10 KPC::RHOA-KO mice). Statistical analysis: one-way ANOVA with post hoc Tukey (D and F), log rank Mantel-Cox test (G), and two-tailed Student’s t test (I): * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Journal: Cell reports

Article Title: Context-dependent inhibitory roles of RhoA in 3D invasive cell migration within the extracellular matrix

doi: 10.1016/j.celrep.2025.116649

Figure Lengend Snippet: (A) Histological analysis (H&E and Masson’s trichrome staining). Scale bar, 200 μm. The enlarged boxed regions highlight details. Scale bar, 100 μm. (B) PDAC progression stages ( n = 7–10 mice for each genotype). (C) Immunohistochemistry of pancreas stained for Ki67 and α-SMA. Scale bars, 20 μm for Ki67 and 100 μm α-SMA. (D) Quantification of Ki67-positive nuclei and α-SMA-positive area (mean ± SD, n = 4 mice for each genotype). (E) Representative pancreas images. Scale bar, 1 cm. (F) Pancreas weight quantification (mean ± SD, n = 6 WT, 7 KPC, 9 KPC::RHOA-KO mice). (G) Survival curve of KPC and KPC::RHOA-KO mice ( n = 10 mice for each genotype). (H) H&E-stained pancreas sections of KPC and KPC::RHOA-KO mice at 12 weeks post-tamoxifen injection. Scale bar, 1 cm. Tumor boundaries are outlined; boxed regions are enlarged. (I) Quantification of inverse circularity of tumors (mean ± SD, n = 20 sections from 6 KPC mice and 50 sections from 10 KPC::RHOA-KO mice). Statistical analysis: one-way ANOVA with post hoc Tukey (D and F), log rank Mantel-Cox test (G), and two-tailed Student’s t test (I): * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: KPC mice , Jackson Laboratory , Strain# 032429.

Techniques: Staining, Immunohistochemistry, Injection, Two Tailed Test

( A ) Representative macroscopic tumor images and tumor weights from KPC Hdac5-WT and KPC Hdac5-KO mouse models treated with vehicle or MRTX1133 (30 mg/kg, i.p., twice daily [bid]) ( n = 5). ( B ) Kaplan-Meier survival curves with log-rank test ( n = 5). ( C and D ) Representative IHC images of tumors in C . IHC scores were quantified in D . Scale bars = 100 μm. n = 5 biologically independent repeats and 3 independent IHC quantifications. ( E ) C57BL/6 mice were orthotopically injected with KPC-Luc cells expressing shNc or shHdac5. Bioluminescence imaging was performed on day 7, followed by treatment with vehicle or MRTX1133 (30 mg/kg, i.p., bid). Representative bioluminescence images and corresponding quantification were acquired on day 28 ( n = 5). ( F ) Tumor growth curves in PDX models with treatment with vehicle or MRTX1133 (30 mg/kg, i.p., bid) ( n = 5). ( G and H ) Representative IHC images of PDXs and quantified IHC scores ( H ). Scale bars = 100 μm. n = 5 biologically independent repeats and 3 independent IHC quantifications. All data are presented as the mean ± SD. Statistical significance was determined by 2-way ANOVA followed by Tukey’s multiple comparisons test ( A , D – F , and H ). * P < 0.05, *** P < 0.001.

Journal: The Journal of Clinical Investigation

Article Title: HDAC5 deficiency induces intrinsic resistance to KRAS inhibition by disrupting c-Myc acetylation-ubiquitination homeostasis

doi: 10.1172/JCI195814

Figure Lengend Snippet: ( A ) Representative macroscopic tumor images and tumor weights from KPC Hdac5-WT and KPC Hdac5-KO mouse models treated with vehicle or MRTX1133 (30 mg/kg, i.p., twice daily [bid]) ( n = 5). ( B ) Kaplan-Meier survival curves with log-rank test ( n = 5). ( C and D ) Representative IHC images of tumors in C . IHC scores were quantified in D . Scale bars = 100 μm. n = 5 biologically independent repeats and 3 independent IHC quantifications. ( E ) C57BL/6 mice were orthotopically injected with KPC-Luc cells expressing shNc or shHdac5. Bioluminescence imaging was performed on day 7, followed by treatment with vehicle or MRTX1133 (30 mg/kg, i.p., bid). Representative bioluminescence images and corresponding quantification were acquired on day 28 ( n = 5). ( F ) Tumor growth curves in PDX models with treatment with vehicle or MRTX1133 (30 mg/kg, i.p., bid) ( n = 5). ( G and H ) Representative IHC images of PDXs and quantified IHC scores ( H ). Scale bars = 100 μm. n = 5 biologically independent repeats and 3 independent IHC quantifications. All data are presented as the mean ± SD. Statistical significance was determined by 2-way ANOVA followed by Tukey’s multiple comparisons test ( A , D – F , and H ). * P < 0.05, *** P < 0.001.

Article Snippet: KP [ Kras tm1(LSL-G12D) Trp53 tm1(LSL-R172H) , C001320] mice, Hdac5 -KO (S-KO-02424) mice, and Tg( Pdx1-Cre ) (C001408) mice were all purchased from Cyagen and interbred to generate KPC Hdac5-KO mice.

Techniques: Injection, Expressing, Imaging

( A ) Venn diagram showing the overlap of genes upregulated in KPC Hdac5-KO mice ( n = 5 per genotype; log 2 [fold-change] > 1, P < 0.05, 2,410 genes) identified via RNA-Seq and genes upregulated in shHDAC5-treated PANC-1 cells ( n = 3 per condition; log 2 [fold-change] > 1, P < 0.05, 2,683 genes), revealing a shared subset of 378 genes. ( B ) Bar graph showing the top 10 enriched transcription factors from transcription factor analysis of 378 genes in A using Enrichr. ATF2, activating transcription factor 2. ( C ) Dual-luciferase reporter assays were performed to assess the transcriptional activity of c-Myc in HDAC5-depleted PANC-1 and AsPC-1 cells. Data are presented as mean ± SD ( n = 5). Statistical significance was determined by 1-way ANOVA followed by Dunnett’s multiple comparisons test. *** P < 0.001. ( D ) Heatmap of MYC ChIP-Seq signal intensity (±3 kb around MYC binding sites) in control vs. HDAC5-knockdown PANC-1 cells. ( E ) The average ChIP signal of MYC centered at transcription start site (±3 kb) in indicated groups. ( F ) Venn diagram depicting the overlap between genes with enhanced MYC promoter occupancy after shHDAC5 knockdown, as identified by ChIP-Seq, and genes upregulated in PANC-1 cells following shHDAC5 knockdown, as determined by RNA-Seq. P = 6 × 10 –71 . ( G ) KEGG pathway enrichment analysis of 821 intersecting genes in F . ( H and I ) Western blot analysis of canonical MAPK pathway protein expression under indicated conditions.

Journal: The Journal of Clinical Investigation

Article Title: HDAC5 deficiency induces intrinsic resistance to KRAS inhibition by disrupting c-Myc acetylation-ubiquitination homeostasis

doi: 10.1172/JCI195814

Figure Lengend Snippet: ( A ) Venn diagram showing the overlap of genes upregulated in KPC Hdac5-KO mice ( n = 5 per genotype; log 2 [fold-change] > 1, P < 0.05, 2,410 genes) identified via RNA-Seq and genes upregulated in shHDAC5-treated PANC-1 cells ( n = 3 per condition; log 2 [fold-change] > 1, P < 0.05, 2,683 genes), revealing a shared subset of 378 genes. ( B ) Bar graph showing the top 10 enriched transcription factors from transcription factor analysis of 378 genes in A using Enrichr. ATF2, activating transcription factor 2. ( C ) Dual-luciferase reporter assays were performed to assess the transcriptional activity of c-Myc in HDAC5-depleted PANC-1 and AsPC-1 cells. Data are presented as mean ± SD ( n = 5). Statistical significance was determined by 1-way ANOVA followed by Dunnett’s multiple comparisons test. *** P < 0.001. ( D ) Heatmap of MYC ChIP-Seq signal intensity (±3 kb around MYC binding sites) in control vs. HDAC5-knockdown PANC-1 cells. ( E ) The average ChIP signal of MYC centered at transcription start site (±3 kb) in indicated groups. ( F ) Venn diagram depicting the overlap between genes with enhanced MYC promoter occupancy after shHDAC5 knockdown, as identified by ChIP-Seq, and genes upregulated in PANC-1 cells following shHDAC5 knockdown, as determined by RNA-Seq. P = 6 × 10 –71 . ( G ) KEGG pathway enrichment analysis of 821 intersecting genes in F . ( H and I ) Western blot analysis of canonical MAPK pathway protein expression under indicated conditions.

Techniques: RNA Sequencing, Luciferase, Activity Assay, ChIP-sequencing, Binding Assay, Control, Knockdown, Western Blot, Expressing

( A ) Representative macroscopic images of pancreatic tumors from KPC Hdac5-WT and KPC Hdac5-KO mice after sacrifice. ( B ) Tumor weight analysis in KPC mice treated with vehicle, MRTX1133 (30 mg/kg, i.p. bid), MYCi975 (50 mg/kg, i.p. bid), or their combination ( n = 5). ( C ) Kaplan-Meier survival curves with log-rank test ( n = 5). *** P < 0.001. ( D and E ) Representative IHC images of tumors from treated mice and quantified IHC scores ( E ). Scale bars = 100 μm. n = 5 biologically independent repeats and 3 independent quantifications. ( F and G ) Representative bioluminescence images and quantification ( n = 5). ( H ) Macroscopic images of PDX tumors after 30 days of treatment. ( I and J ) Tumor growth curves ( I ) and final tumor weights ( J ) in PDX models treated with Vehicle + LV-Control, MRTX1133 (30 mg/kg, i.p. bid) + LV-Control, Vehicle + LV-sgMYC (50 μL lentivirus, s.c., weekly), or MRTX1133 + LV-sgMYC. All data are presented as the mean ± SD. Statistical significance was determined by 2-way ANOVA followed by Tukey’s multiple comparisons test ( B , E , G , I , and J ). *** P < 0.001.

Journal: The Journal of Clinical Investigation

Article Title: HDAC5 deficiency induces intrinsic resistance to KRAS inhibition by disrupting c-Myc acetylation-ubiquitination homeostasis

doi: 10.1172/JCI195814

Figure Lengend Snippet: ( A ) Representative macroscopic images of pancreatic tumors from KPC Hdac5-WT and KPC Hdac5-KO mice after sacrifice. ( B ) Tumor weight analysis in KPC mice treated with vehicle, MRTX1133 (30 mg/kg, i.p. bid), MYCi975 (50 mg/kg, i.p. bid), or their combination ( n = 5). ( C ) Kaplan-Meier survival curves with log-rank test ( n = 5). *** P < 0.001. ( D and E ) Representative IHC images of tumors from treated mice and quantified IHC scores ( E ). Scale bars = 100 μm. n = 5 biologically independent repeats and 3 independent quantifications. ( F and G ) Representative bioluminescence images and quantification ( n = 5). ( H ) Macroscopic images of PDX tumors after 30 days of treatment. ( I and J ) Tumor growth curves ( I ) and final tumor weights ( J ) in PDX models treated with Vehicle + LV-Control, MRTX1133 (30 mg/kg, i.p. bid) + LV-Control, Vehicle + LV-sgMYC (50 μL lentivirus, s.c., weekly), or MRTX1133 + LV-sgMYC. All data are presented as the mean ± SD. Statistical significance was determined by 2-way ANOVA followed by Tukey’s multiple comparisons test ( B , E , G , I , and J ). *** P < 0.001.

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1) Product Images from "The Lysine Demethylase KDM4C Is an Oncogenic Driver and Regulates ERK Activity in KRAS-Mutant Pancreatic Ductal Adenocarcinoma"

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