Journal: bioRxiv

Article Title: Spt5 interacts genetically with Myc and is limiting for brain tumor growth in Drosophila

doi: 10.1101/2023.04.14.536839

Figure Lengend Snippet: A , number of NB II-derived (GFP-positive) cells per larva (based on 3-7 independent dissections per genotype, each involving 130-240 larvae). B , NB II lineages in brains from 3rd instar larvae were marked with mCD8::GFP (green) and co-stained for the nuclear proteins Dpn (red) and the apoptosis marker Dcp-1 (blue). Few apoptotic cells are observed in control, Spt5-KD and brat-KD brains but their number increases under double-knockdown conditions. Scale bar: 10 µm.

Article Snippet: After blocking in PBT containing 5% normal goat serum for 1 h, tissues were incubated overnight at 4°C with combinations of the following primary antibodies: rabbit anti-Ase (1:400; F. Diaz-Benjumea, Madrid, Spain), mouse anti-Bruchpilot (1:30, clone nc82; E. Buchner, Würzburg, Germany), rabbit anti-Dcp-1 (1:100, Cell Signaling Techn.

Techniques: Derivative Assay, Staining, Marker

Journal: Frontiers in Cell and Developmental Biology

Article Title: Integrins Cooperate With the EGFR/Ras Pathway to Preserve Epithelia Survival and Architecture in Development and Oncogenesis

doi: 10.3389/fcell.2022.892691

Figure Lengend Snippet: Integrins promote cell survival in the wing imaginal disc. (A–D′′) Maximal projection of confocal views of wing imaginal discs from third-instar larvae stained with anti-βPS [green in (A–D) , white in (A′–D′)] , anti-Dcp1 [red in (A–D) , white in (A′′–D′′)] and the nuclear marker Hoechst [DNA, blue in (A–D) ]. (A) Control nubGal4 wing disc. (B) Wing disc carrying mys mutant clones (white arrow), marked by the absence of βPS (green). (C) Wing disc expressing GFP or a mys RNAi under the control of nubGal4 , nub > GFP (inset) and nub > mys RNAi , respectively. (yz) Confocal yz section along the white dotted line shown in (C) . (D) Wing disc co-expressing a mys RNAi and Diap-1 under the control of nubGal4 , nub > mys RNAi ; Diap1 . (E) Quantification of Dcp1 levels in wing discs of the designated genotypes. The dotted white circles indicate the wing pouch area in all figures. The statistical significance of differences was assessed with a t -test, ***, ** and * p values < 0.001, <0.01, and <0.05, respectively. Scale bar in all panels, 30 μm.

Article Snippet: The following primary antibodies were used: goat anti-GFP FICT (Abcam, 1:500), rabbit anti-caspase Dcp1 (Cell Signaling; 1:100), rabbit anti-pJNK (Promega, 1:200), mouse anti-βGal (Promega, 1:1000), mouse anti-βPS (Developmental Studies Hybridoma Bank, DSHB, University of Iowa, United States, 1:50) and mouse anti-myc (Promega, 1:100).

Techniques: Staining, Marker, Control, Mutagenesis, Clone Assay, Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: Integrins Cooperate With the EGFR/Ras Pathway to Preserve Epithelia Survival and Architecture in Development and Oncogenesis

doi: 10.3389/fcell.2022.892691

Figure Lengend Snippet: Integrins stimulate cell survival through inhibition of hid expression. (A–D‴) Maximal projection of confocal views of third-instar wing imaginal discs stained with anti-βPS [blue in (A–D) , white in (A′–D′)] , anti-Dcp1 [red in (A–D) , white in (A′′–D′′)] and anti-GFP [green in (A–D) , white in (A‴–D‴)] . (A–A‴) Control nubGal4 wing disc carrying a reporter of hid expression, hidGFP , hidGFP; nubGal4 . (B–B‴) hidGFP wing disc expressing a mys RNAi under the control of nubGal4 , hidGFP;nub > mys RNAi . (C–D‴) hidGFP wing disc co-expressing a mys RNAi under the control of nubGal4 with either Diap1, hidGFP; nub > mys RNAi ; Diap1 (C–C‴) , or a hid RNAi, hidGFP;nub > mys RNAi ; hid RNAi (D–D‴) . (E) Quantification of Dcp1 levels in wing discs of the indicated genotypes. The statistical significance of differences was assessed with a t -test, ***, ** and * p values < 0.001, <0.01, and <0.05, respectively. NS, non-statistically significant. Scale bar in all panels, 30 μm.

Article Snippet: The following primary antibodies were used: goat anti-GFP FICT (Abcam, 1:500), rabbit anti-caspase Dcp1 (Cell Signaling; 1:100), rabbit anti-pJNK (Promega, 1:200), mouse anti-βGal (Promega, 1:1000), mouse anti-βPS (Developmental Studies Hybridoma Bank, DSHB, University of Iowa, United States, 1:50) and mouse anti-myc (Promega, 1:100).

Techniques: Inhibition, Expressing, Staining, Control

Journal: Frontiers in Cell and Developmental Biology

Article Title: Integrins Cooperate With the EGFR/Ras Pathway to Preserve Epithelia Survival and Architecture in Development and Oncogenesis

doi: 10.3389/fcell.2022.892691

Figure Lengend Snippet: Integrins promote cell survival by inhibiting the JNK pathway. (A–C‴) Maximal projection of confocal views of third-instar wing imaginal discs carrying the reporter hidGFP , stained with anti-βPS [blue in (A–C) , white in (A′–C′)] , anti-Dcp1 [red in (A–C) , white in (A′′–C′′)] and anti-GFP [green in (A–C) , white in (A‴–C‴)] . (A–A‴) Control hidGFP ; nubGal4 wing disc. (B–B‴) hidGFP wing disc expressing a mys RNAi under the control of nubGal4 , hidGFP ; nub > mys RNAi . (C–C‴) hidGFP wing disc co-expressing a mys RNAi and the negative regulator of the JNK pathway puc , under the control of nubGal4 hidGFP ; nub > mys RNAi ; puc . (D,E) Quantification of Dcp1 (D) and hidGFP (E) levels in wing discs of the indicated genotypes. The statistical significance of differences was assessed with a t -test, *** and * p values < 0.001, <0.01, and <0.05, respectively. NS, non-statistically significant. Scale bar in all panels, 30 μm.

Article Snippet: The following primary antibodies were used: goat anti-GFP FICT (Abcam, 1:500), rabbit anti-caspase Dcp1 (Cell Signaling; 1:100), rabbit anti-pJNK (Promega, 1:200), mouse anti-βGal (Promega, 1:1000), mouse anti-βPS (Developmental Studies Hybridoma Bank, DSHB, University of Iowa, United States, 1:50) and mouse anti-myc (Promega, 1:100).

Techniques: Staining, Control, Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: Integrins Cooperate With the EGFR/Ras Pathway to Preserve Epithelia Survival and Architecture in Development and Oncogenesis

doi: 10.3389/fcell.2022.892691

Figure Lengend Snippet: Integrins and EGFR cooperate to promote cell survival. (A–D”) Maximal projection of confocal views of wing imaginal discs from third-instar larvae carrying the reporter hidGFP , stained with anti-Dcp1 [red in (A–D) , white in (A′–D′)] , anti-GFP [green in (A–D) , white in (A’’–D’’)] and the nuclear marker Hoechst [DNA, blue in (A–D) ]. (A–D’’) Control hidGFP ; nubGal4 wing disc. (B–B’’) hidGFP wing disc expressing a mys RNAi under the control of nubGal4 , hidGFP;nub > mys RNAi . (C–C’’) hidGFP wing disc expressing a dominant negative form of the EGFR, DER DN , under the control of nubGal4 , hidGFP ; nub > DER DN . (C–D’’) hidGFP wing disc co-expressing a mys RNAi and DER DN under the control of nubGal4 hidGFP ; nub > mys RNAi ; DER DN . (E) Quantification of Dcp1 levels in wing discs of the indicated genotypes. The statistical significance of differences was assessed with a t -test, *** and ** p values < 0.001 and <0.01, respectively. NS, non-statistically significant. Scale bar in all panels, 30 μm.

Article Snippet: The following primary antibodies were used: goat anti-GFP FICT (Abcam, 1:500), rabbit anti-caspase Dcp1 (Cell Signaling; 1:100), rabbit anti-pJNK (Promega, 1:200), mouse anti-βGal (Promega, 1:1000), mouse anti-βPS (Developmental Studies Hybridoma Bank, DSHB, University of Iowa, United States, 1:50) and mouse anti-myc (Promega, 1:100).

Techniques: Staining, Marker, Control, Expressing, Dominant Negative Mutation

Journal: Frontiers in Cell and Developmental Biology

Article Title: Integrins Cooperate With the EGFR/Ras Pathway to Preserve Epithelia Survival and Architecture in Development and Oncogenesis

doi: 10.3389/fcell.2022.892691

Figure Lengend Snippet: Ectopic activation of the Ras pathway rescues anoikis. (A–C’’’) Maximal projection of confocal views of third-instar wing imaginal discs carrying the reporter hidGFP , stained with anti-βPS [blue in (A–C) , white in (A′–C′)] , anti-Dcp1 [red in (A–C) , white in (A’’–C’’)] and anti-GFP [green in (A–C) , white in (A‴–C‴)] . (A–A‴) Control hidGFP;nubGal4 wing disc. (B–B‴) hidGFP wing disc expressing a mys RNAi under the control of nubGal4 , hidGFP ; nub > mys RNAi . (C–C‴) hidGFP wing disc co-expressing a mys RNAi and an activated form of Ras, Ras V12 , under the control of nubGal4 hidGFP ; nub > mys RNAi ; Ras V12 . (D) Quantification of Dcp1 levels in wing discs of the indicated genotypes. The statistical significance of differences was assessed with a t -test, *** and * p values < 0.001 and <0.05, respectively. Scale bar in all panels, 30 μm.

Article Snippet: The following primary antibodies were used: goat anti-GFP FICT (Abcam, 1:500), rabbit anti-caspase Dcp1 (Cell Signaling; 1:100), rabbit anti-pJNK (Promega, 1:200), mouse anti-βGal (Promega, 1:1000), mouse anti-βPS (Developmental Studies Hybridoma Bank, DSHB, University of Iowa, United States, 1:50) and mouse anti-myc (Promega, 1:100).

Techniques: Activation Assay, Staining, Control, Expressing