ptripz shns  (Addgene inc)

Journal: Oncogene

Article Title: Drug tolerance and persistence to EGFR inhibitor treatment are mediated by an ILK-SFK-YAP signaling axis in lung adenocarcinoma.

doi: 10.1038/s41388-025-03461-6

Figure Lengend Snippet: Fig. 3 ILK knockdown improves response to Osi treatment. a Schematic of pTRIPZ shRNA constructs. b Validation of knockdown in HCC4006 shILK cells; western blot of proteins in the IPP complex in HCC4006. c Three day Osi dose-response in HCC4006 shILK (n = 3) and shNS (n = 3) ± DOX. Multiple t-tests with Holm-Sidak correction were used to statistically compare DOX vs No DOX at each Osi concentration (*p < 0.05). d Clonogenic assay of HCC4006 shILK (n = 10) and shNS (n = 9) cells treated with Osi for 7 days ± DOX. Quantification of colony count is below the representative plates. Multiple paired t-tests with Holm-Sidak correction were used to statistically compare DOX vs No DOX at each Osi concentration (*p < 0.05; **p < 0.01). Error bars represent the mean ± SEM. DOX concentration = 100 ng/mL.

Article Snippet: HEK293T cells co-transfected with pLenti CMV ILK Blast, pTRIPZ shNS (a gift from Sandra Demaria; Addgene plasmid # 127696), pTRIPz shILK B09, pLenti CMV GFP Blast (a gift from Eric Campeau & Paul Kaufman; Addgene plasmid # 17445), EF1a_mCherry_P2A_Hygro (a gift from Prashant Mali; Addgene plasmid # 135003), pInducer20-SRC, or Oncogene pInducer20-GFP plasmids (as described previously [45]) together with pMD2.G, and psPAX2 plasmid constructs (pMD2.G and psPAX2 were gifts from Didier Trono; Addgene plasmid #12259 and #12260, respectively) using Lipofectamine 2000 (Thermo Fisher Scientific).

Techniques: Knockdown, shRNA, Construct, Biomarker Discovery, Western Blot, Concentration Assay, Clonogenic Assay

Journal: Oncogene

Article Title: Drug tolerance and persistence to EGFR inhibitor treatment are mediated by an ILK-SFK-YAP signaling axis in lung adenocarcinoma.

doi: 10.1038/s41388-025-03461-6

Figure Lengend Snippet: Fig. 4 ILK promotes DTP survival and the progression towards EMT-mediated Osi resistance. a Western blot of EMT markers in HCC4006 shNS, and shILK cells ± DOX Par and OsiR. b Western blot of ILK and EMT markers following treatment with DMSO or 100 nM Osi for 7 days and subsequent removal of Osi. c Schematic of persister cell assay. d Persister clonogenic assay of HCC4006 shILK (n = 6) and shNS (n = 6) cells treated with Osi for 7 days ± DOX. Quantification of colony count is below the representative plates. Multiple paired t-tests with Holm-Sidak correction were used to statistically compare DOX vs No DOX at each Osi concentration (*p < 0.05; **p < 0.01). Error bars represent the mean ± SEM. DOX concentration = 100 ng/mL.

Article Snippet: HEK293T cells co-transfected with pLenti CMV ILK Blast, pTRIPZ shNS (a gift from Sandra Demaria; Addgene plasmid # 127696), pTRIPz shILK B09, pLenti CMV GFP Blast (a gift from Eric Campeau & Paul Kaufman; Addgene plasmid # 17445), EF1a_mCherry_P2A_Hygro (a gift from Prashant Mali; Addgene plasmid # 135003), pInducer20-SRC, or Oncogene pInducer20-GFP plasmids (as described previously [45]) together with pMD2.G, and psPAX2 plasmid constructs (pMD2.G and psPAX2 were gifts from Didier Trono; Addgene plasmid #12259 and #12260, respectively) using Lipofectamine 2000 (Thermo Fisher Scientific).

Techniques: Western Blot, Clonogenic Assay, Concentration Assay

Journal: Oncogene

Article Title: Drug tolerance and persistence to EGFR inhibitor treatment are mediated by an ILK-SFK-YAP signaling axis in lung adenocarcinoma.

doi: 10.1038/s41388-025-03461-6

Figure Lengend Snippet: Fig. 7 ILK preserves residual tumours and promotes YAP activation during Osi treatment in vivo. a Schematic of HCC4006 shNS/shILK tumour xenograft experiment. b Validation of ILK knockdown by RT-qPCR. Ratio paired t-test was used to statistically compare between HCC4006 shNS vs shILK tumours (n = 7) in the same mouse (***p < 0.001). c Tumour weights of implanted HCC4006 shNS and shILK tumours extracted from mice gavaged with Vehicle (Veh; n = 4) or Osi (n = 3) at endpoint. Ratio paired t-test was used to statistically compare between HCC4006 shNS vs shILK tumours (n = 3) in the same mouse (**p < 0.01). d Representative western blot of HCC4006 shNS and shILK tumours treated with Veh or Osi. Quantification of YAP band density are plotted on the right. Ratio paired t-test was used to statistically compare between HCC4006 shNS vs shILK tumours (n = 3) in the same mouse (*p < 0.05). e Representative images of YAP staining in HCC4006 shNS and shILK tumours. White arrows indicate regions of YAP staining. f Quantification of nuclear YAP intensity and %YAP positive area between shNS vs shILK tumours (n = 4 images/section × 2 section/tumour × 3 tumours). Student’s t-tests were used to statistically compare HCC4006 shNS vs shILK persistent tumours (*p < 0.05, ***p < 0.001). Error bars represent the mean ± SEM.

Article Snippet: HEK293T cells co-transfected with pLenti CMV ILK Blast, pTRIPZ shNS (a gift from Sandra Demaria; Addgene plasmid # 127696), pTRIPz shILK B09, pLenti CMV GFP Blast (a gift from Eric Campeau & Paul Kaufman; Addgene plasmid # 17445), EF1a_mCherry_P2A_Hygro (a gift from Prashant Mali; Addgene plasmid # 135003), pInducer20-SRC, or Oncogene pInducer20-GFP plasmids (as described previously [45]) together with pMD2.G, and psPAX2 plasmid constructs (pMD2.G and psPAX2 were gifts from Didier Trono; Addgene plasmid #12259 and #12260, respectively) using Lipofectamine 2000 (Thermo Fisher Scientific).

Techniques: Activation Assay, In Vivo, Biomarker Discovery, Knockdown, Quantitative RT-PCR, Western Blot, Staining