martine roussel  (Addgene inc)

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Optogenetic clustering and membrane translocation of the BcLOV4 photoreceptor.

doi: 10.1073/pnas.2221615120

Figure Lengend Snippet: Fig. 3. BcLOV4 clustering at the membrane allows for modular activation of the entire ErbB receptor family. (A) The intracellular domains of ErbB1-4 were fused to the C-terminus of BcLOV-mCherry. (B) Membrane translocation of BcLOV-ErbB1-4 in response to blue light. (Scale bar, 20 μm.) See also Movie S3. (C and D) Erk activation dynamics downstream of BcLOV-ErbB1-4 in response to ON and OFF steps of blue light. See also SI Appendix, Fig. S4. Data represent mean ± SEM of two replicates, with each replicate representing the mean of ~500 to 2,400 cells. (E) Membrane ruffling (black arrows) downstream of stimulation of BcLOV- ErbB1-4, indicative of RTK stimulation. Ruffling is strongest for ErbB1 and ErbB2, less for ErbB4, and absent for ErbB3 activation. (Scale bar, 20 μm.) See also Movie S4. (F) Kinase activity suppresses BcLOV-EGFR membrane translocation. Translocation was observed under light stimulation of BcLOV-EGFR harboring a kinase-inactivating D813N mutation, in either the presence or absence of 1 µM EGFR inhibitor erlotinib (EGFRi). (Scale bar, 20 μm.) Quantification (Right) shows ratios of mean membrane and cytoplasmic fluorescence of 15 cells per condition. Significance was tested by one-way t tests between individual groups and was assessed by comparing P values to Bonferroni-corrected significance level of α/3. ****P < 0.00001, ***P < 0.0001.

Article Snippet: ErbB2 was sourced from MSCV- human Erbb2- IRES- GFP, which was a gift from Martine Roussel (Addgene plasmid # 91888; http://n2t.net/ addgene:91888; RRID:Addgene_91888).

Techniques: Membrane, Activation Assay, Translocation Assay, Activity Assay, Mutagenesis, Fluorescence

Journal: Cancers

Article Title: Potent Antitumor Activity of Liposomal Irinotecan in an Organoid- and CRISPR-Cas9-Based Murine Model of Gallbladder Cancer

doi: 10.3390/cancers11121904

Figure Lengend Snippet: Mutant ERBB2 cooperates with loss of p53 and leads to papillary GBC in recipient mice. ( A ) Top: Schematic of human ERBB2, indicating the location of two point mutants (S310F and V777L). Bottom: retroviral vector used to transduce organoids, that had been treated with an sgp53-containing plasmid (px459) to induce loss of p53. ( B ) Immunofluorescence for ERBB2 (top) and phospho-ERBB2 (bottom) on organoids harboring the indicated genetic alterations. ( C ) Tumor volumes 36 days after s.c. implantation of the respective organoids into recipient mice. All mice transplanted with sgp53;ERBB2 S310F - and sgp53;ERBB2 V777L organoids exhibited tumor development, whereas sgp53;empty vector- and sgp53;ERBB2 wildtype organoids did not give rise to tumors over a four-month observation period. There was no significant difference in the tumor burden of mice transplanted with sgp53;ERBB2 S310F - and sgp53;ERBB2 V777L organoids ( p = 0.999). ( D ) Mice transplanted with sgp53;ERBB2 S310F - and sgp53;ERBB2 V777L organoids reached endpoint criteria with a median survival of 79.5 days and 58.5 days, respectively. ( E ) H&E and IHC for CK19 and EGFP on tumors generated with sgp53;ERBB2 S310 - and sgp53;ERBB2 V777L organoids.

Article Snippet: The pMSCV- ERBB2 -IRES- EGFP was a gift from Martine Roussel (Addgene, Watertown, MA, USA, plasmid #91888).

Techniques: Mutagenesis, Retroviral, Plasmid Preparation, Transduction, Immunofluorescence, Generated