other:
Incubation:Article Title: Matrix stiffness induces endothelial network senescence
Article Snippet: .. Membranes were then incubated overnight at 4 °C with primary antibodies targeting: p21 (Cell Signaling, rabbit, 1:1000), NICD (Cell Signaling, rabbit, 1:1000), JAG1 (Cell Signaling, rabbit, 1:1000), JAG2 (Cell Signaling, rabbit, 1:1000), phospho-JNK (Cell Signaling, rabbit, 1:750), JNK (Cell Signaling, rabbit, 1:750), phospho-cJUN (Cell Signaling, rabbit, 1:750), cJUN (Cell Signaling, rabbit, 1:750), and PCNA (Proteintech, rabbit, 1:1000). .. Following primary antibody incubation, blots were washed three times with TBST and incubated with species-appropriate HRP-conjugated secondary antibodies or HRP-conjugated GAPDH (Cell Signaling, rabbit, 1:2500).
Binding Assay:Article Title: NAD + sensing by PARP7 regulates the C/EBPβ-dependent transcription program during adipogenesis.
Article Snippet: .. Other antibodies used were as follows: PARP7 (Thermo Fisher Scientific, PA5-40774; RRID:AB_2607074), C/EBPβ (Invitrogen, PA5-120052; RRID:AB_2913624, Invitrogen, PA5-86117; RRID:AB_2802916, and Cell Signaling Technology, 3082; RRID:AB_2260365), Phospho-C/EBPβ (Thr235) (Cell Signaling Technology, 3084S; RRID:AB_2260359), PPARγ (81B8) (Cell Signaling Technology, 2443; RRID:AB_823598), cJun (Proteintech, 24909-1-AP; RRID:AB_2860574), cJun (Cell Signaling Technology, 9165; RRID:AB_2130165), DTX2 (ThermoFisher, PA5-109664; RRID:AB_2855075), RNF114 (Proteintech, 14338-1-AP; RRID:AB_3085435), Histone H3K27ac (Active Motif, 39134; RRID:AB_2722569), Histone H2BK5ac (Cell Signaling Technology, 12799; RRID:AB_2636805), Histone H3K27me3 (Cell Signaling Technology, 9733; RRID:AB_2616029), FLAG (Sigma-Aldrich, F3165; RRID:AB_259529), MAR binding reagent (Millipore, MABE1076; RRID:AB_2665469), PAR binding reagent (Millipore, MABE1031; RRID:AB_2665467), Perilipin (Biosynth, 20R-PP004; RRID: AB_3665667), Ubiquitin (E4I2J) (Cell Signaling Technology, 43124; RRID:AB_2799235), K48-linkage specific polyubiquitin (Cell Signaling Technology, 4289; RRID:AB_10557239), p300 (Active Motif, 61404; RRID:AB_2716754), p300 (Santa Cruz, SC-584; RRID:AB_2293429), NAMPT (D1K6D) (Cell Signaling Technology, 61122; RRID:AB_2799602), NAMPT (Proteintech, 11776-1-AP; RRID:AB_2298317), β-tubulin (Abcam, ab6046; RRID:AB_2210370), and β-Actin (8H10D10) (Cell Signaling Technology, 3700; RRID:AB_2242334). .. Secondary antibodies included Goat anti-rabbit HRPconjugated IgG (ThermoFisher, 31460; RRID:AB_228341), Goat anti-mouse HRP-conjugated IgG (ThermoFisher, 31430; RRID:AB_10960845), Rabbit IgG (ThermoFisher, 10500C; RRID:AB_2532981), and Alexa Fluor 594 donkey anti-rabbit IgG (ThermoFisher, A-21207; RRID:AB_141637).
Article Title: NAD + sensing by PARP7 regulates the C/EBPβ-dependent transcription program during adipogenesis
Article Snippet: .. Other antibodies used were as follows: PARP7 (Thermo Fisher Scientific, PA5–40774; RRID:AB_2607074), C/EBPβ (Invitrogen, PA5–120052; RRID:AB_2913624, Invitrogen, PA5–86117; RRID:AB_2802916, and Cell Signaling Technology, 3082; RRID:AB_2260365), Phospho-C/EBPβ (Thr235) (Cell Signaling Technology, 3084S; RRID:AB_2260359), PPARγ (81B8) (Cell Signaling Technology, 2443; RRID:AB_823598), cJun (Proteintech, 24909–1-AP; RRID:AB_2860574), cJun (Cell Signaling Technology, 9165; RRID:AB_2130165), DTX2 (ThermoFisher, PA5–109664; RRID:AB_2855075), RNF114 (Proteintech, 14338–1-AP; RRID:AB_3085435), Histone H3K27ac (Active Motif, 39134; RRID:AB_2722569), Histone H2BK5ac (Cell Signaling Technology, 12799; RRID:AB_2636805), Histone H3K27me3 (Cell Signaling Technology, 9733; RRID:AB_2616029), FLAG (Sigma-Aldrich, F3165; RRID:AB_259529), MAR binding reagent (Millipore, MABE1076; RRID:AB_2665469), PAR binding reagent (Millipore, MABE1031; RRID:AB_2665467), Perilipin (Biosynth, 20R-PP004; RRID: AB_3665667), Ubiquitin (E4I2J) (Cell Signaling Technology, 43124; RRID:AB_2799235), K48-linkage specific polyubiquitin (Cell Signaling Technology, 4289; RRID:AB_10557239), p300 (Active Motif, 61404; RRID:AB_2716754), p300 (Santa Cruz, SC-584; RRID:AB_2293429), NAMPT (D1K6D) (Cell Signaling Technology, 61122; RRID:AB_2799602), NAMPT (Proteintech, 11776–1-AP; RRID:AB_2298317), β-tubulin (Abcam, ab6046; RRID:AB_2210370), and β-Actin (8H10D10) (Cell Signaling Technology, 3700; RRID:AB_2242334). .. Secondary antibodies included Goat anti-rabbit HRP-conjugated IgG (ThermoFisher, 31460; RRID:AB_228341), Goat anti-mouse HRP-conjugated IgG (ThermoFisher, 31430; RRID:AB_10960845), Rabbit IgG (ThermoFisher, 10500C; RRID:AB_2532981), and Alexa Fluor 594 donkey anti-rabbit IgG (ThermoFisher, A-21207; RRID:AB_141637).
Ubiquitin Proteomics:Article Title: NAD + sensing by PARP7 regulates the C/EBPβ-dependent transcription program during adipogenesis.
Article Snippet: .. Other antibodies used were as follows: PARP7 (Thermo Fisher Scientific, PA5-40774; RRID:AB_2607074), C/EBPβ (Invitrogen, PA5-120052; RRID:AB_2913624, Invitrogen, PA5-86117; RRID:AB_2802916, and Cell Signaling Technology, 3082; RRID:AB_2260365), Phospho-C/EBPβ (Thr235) (Cell Signaling Technology, 3084S; RRID:AB_2260359), PPARγ (81B8) (Cell Signaling Technology, 2443; RRID:AB_823598), cJun (Proteintech, 24909-1-AP; RRID:AB_2860574), cJun (Cell Signaling Technology, 9165; RRID:AB_2130165), DTX2 (ThermoFisher, PA5-109664; RRID:AB_2855075), RNF114 (Proteintech, 14338-1-AP; RRID:AB_3085435), Histone H3K27ac (Active Motif, 39134; RRID:AB_2722569), Histone H2BK5ac (Cell Signaling Technology, 12799; RRID:AB_2636805), Histone H3K27me3 (Cell Signaling Technology, 9733; RRID:AB_2616029), FLAG (Sigma-Aldrich, F3165; RRID:AB_259529), MAR binding reagent (Millipore, MABE1076; RRID:AB_2665469), PAR binding reagent (Millipore, MABE1031; RRID:AB_2665467), Perilipin (Biosynth, 20R-PP004; RRID: AB_3665667), Ubiquitin (E4I2J) (Cell Signaling Technology, 43124; RRID:AB_2799235), K48-linkage specific polyubiquitin (Cell Signaling Technology, 4289; RRID:AB_10557239), p300 (Active Motif, 61404; RRID:AB_2716754), p300 (Santa Cruz, SC-584; RRID:AB_2293429), NAMPT (D1K6D) (Cell Signaling Technology, 61122; RRID:AB_2799602), NAMPT (Proteintech, 11776-1-AP; RRID:AB_2298317), β-tubulin (Abcam, ab6046; RRID:AB_2210370), and β-Actin (8H10D10) (Cell Signaling Technology, 3700; RRID:AB_2242334). .. Secondary antibodies included Goat anti-rabbit HRPconjugated IgG (ThermoFisher, 31460; RRID:AB_228341), Goat anti-mouse HRP-conjugated IgG (ThermoFisher, 31430; RRID:AB_10960845), Rabbit IgG (ThermoFisher, 10500C; RRID:AB_2532981), and Alexa Fluor 594 donkey anti-rabbit IgG (ThermoFisher, A-21207; RRID:AB_141637).
Article Title: NAD + sensing by PARP7 regulates the C/EBPβ-dependent transcription program during adipogenesis
Article Snippet: .. Other antibodies used were as follows: PARP7 (Thermo Fisher Scientific, PA5–40774; RRID:AB_2607074), C/EBPβ (Invitrogen, PA5–120052; RRID:AB_2913624, Invitrogen, PA5–86117; RRID:AB_2802916, and Cell Signaling Technology, 3082; RRID:AB_2260365), Phospho-C/EBPβ (Thr235) (Cell Signaling Technology, 3084S; RRID:AB_2260359), PPARγ (81B8) (Cell Signaling Technology, 2443; RRID:AB_823598), cJun (Proteintech, 24909–1-AP; RRID:AB_2860574), cJun (Cell Signaling Technology, 9165; RRID:AB_2130165), DTX2 (ThermoFisher, PA5–109664; RRID:AB_2855075), RNF114 (Proteintech, 14338–1-AP; RRID:AB_3085435), Histone H3K27ac (Active Motif, 39134; RRID:AB_2722569), Histone H2BK5ac (Cell Signaling Technology, 12799; RRID:AB_2636805), Histone H3K27me3 (Cell Signaling Technology, 9733; RRID:AB_2616029), FLAG (Sigma-Aldrich, F3165; RRID:AB_259529), MAR binding reagent (Millipore, MABE1076; RRID:AB_2665469), PAR binding reagent (Millipore, MABE1031; RRID:AB_2665467), Perilipin (Biosynth, 20R-PP004; RRID: AB_3665667), Ubiquitin (E4I2J) (Cell Signaling Technology, 43124; RRID:AB_2799235), K48-linkage specific polyubiquitin (Cell Signaling Technology, 4289; RRID:AB_10557239), p300 (Active Motif, 61404; RRID:AB_2716754), p300 (Santa Cruz, SC-584; RRID:AB_2293429), NAMPT (D1K6D) (Cell Signaling Technology, 61122; RRID:AB_2799602), NAMPT (Proteintech, 11776–1-AP; RRID:AB_2298317), β-tubulin (Abcam, ab6046; RRID:AB_2210370), and β-Actin (8H10D10) (Cell Signaling Technology, 3700; RRID:AB_2242334). .. Secondary antibodies included Goat anti-rabbit HRP-conjugated IgG (ThermoFisher, 31460; RRID:AB_228341), Goat anti-mouse HRP-conjugated IgG (ThermoFisher, 31430; RRID:AB_10960845), Rabbit IgG (ThermoFisher, 10500C; RRID:AB_2532981), and Alexa Fluor 594 donkey anti-rabbit IgG (ThermoFisher, A-21207; RRID:AB_141637).
Western Blot:Article Title: Mutant RIT1 cooperates with YAP to drive an EMT-like lung cancer state
Article Snippet: Quantification of Western blot band intensity was performed using Licor Empiria Studio with normalization to the loading control. .. Primary antibodies used for immunoblotting: β-Actin 1:10000 (Cell Signaling Technology, 4970; Proteintech, 66009-1-Ig), Vinculin 1:10000 (Sigma, V9264), RIT1 1:2000 (Abcam, Ab53720), YAP (D8H1X) 1:200 (Cell Signaling Technology), cJUN 1:1000 (Cell Signaling Technology, 9165). .. FRA1 (D80B4) (Cell Signaling Technology, 5281), HA-Tag (C29F4) (3724, Cell Signaling Technology), ZEB1 (Abcam, ab155249), DYKDDDDK-Tag 1:500 (14793, Cell Signaling Technology).
Control:Article Title: Mutant RIT1 cooperates with YAP to drive an EMT-like lung cancer state
Article Snippet: .. The following antibodies were used: YAP1 (Abcam #ab52771); TAZ (CST #4883); panTEAD (CST #13295); cJUN (CST #9165); FOSL1 (Abcam #ab252421); H3K4me3 (CST #9751); H3K4me1 (CST #5326); H3K27me3 (CST #9733); H3K27ac (CST #8173); Pol II (CST #13523); ZEB1 (Abcam #ab155249); IgG control (CST #3900). ..
Article Title: Mutant RIT1 cooperates with YAP to drive an EMT-like lung cancer state.
Article Snippet: .. The following antibodies were used: YAP1 (Abcam #ab52771); TAZ (CST #4883); panTEAD (CST #13295); cJUN (CST #9165); FOSL1 (Abcam #ab252421); H3K4me3 (CST #9751); H3K4me1 (CST #5326); H3K27me3 (CST #9733); H3K27ac (CST #8173); Pol II (CST #13523); ZEB1 (Abcam #ab155249); IgG control (CST #3900). .. Primary antibodies were added to a final dilution of 1:100 and incubated for 2hrs rotating at 4 ◦ C. Samples were processed for AutoCUT&RUN by arraying in a 96 well PCR plate and processed on a Beckman Biomek i7 liquid handling instrument equipped with a 96S Super Magnet Plate (Alpaqua SKU A001322) for magnetic separation of samples during wash steps, and a QInstruments ColdPlate heater-cooler thermo block (QInstruments no. 2016-0110) with a 96 well PCR plate adapter (Qinstruments no. 2016-1041) to perform the MNase digestion reaction at 4 ◦ C after the addition of 2 mM CaCl2.
|
Cell Signaling Technology Inc is a verified supplier 
Cell Signaling Technology Inc manufactures this product 
![A subpopulation of RGCs express the stress marker phosphorylated c-JUN <t>(p-cJUN).</t> (A) Whole mount image ( Cx3cr1-Gfp/+ ) of e14.5 retina showing that a subset of RGCs (Brn3a+; red) express nuclear p-cJUN (white; yellow arrows). Boxed regions show enlarged region and label for p-cJUN alone or Brn3a alone (A′) . Scale bar = 10 μm. (B) Whole mount image (e14.5 Cx3cr1-Gfp/+ ) showing microglia (GFP) internalizing an RGC (Brn3a + red) labeled with p-cJUN (white) using HCR IHC. Scale bar = 1 μm. (C) Whole mount images showing RGCs (Brn3a+; red) with nuclear p-cJUN (white) in control mice (WT) and Mertk/CR3 dKO mice. Yellow arrows indicate double positive p-cJUN and Brn3a staining. (D) Quantification of double positive p-cJUN + Brn3a + RGCs per mm 2 in e14.5 control and Mertk/CR3 dKO mice. *** p < 0.01 unpaired t -test [ N = 4 (WT), 5 (Mertk CR3 dKO)]. (E) Quantification of p-cJUN + Brn3a + RGCs per mm 2 in e14.5 and e16.5 Mertk/CR3 dKO mice. * p < 0.05 Welch’s t -test [ N = 5 (e14.5), 4 (e16.5)]. (F) Quantification of p-cJUN + Brn3a + RGCs per mm 2 in P0 control, Mertk KO and Mertk/CR3 dKO mice. ns, not significant. Welch’s one-way ANOVA with Brown–Forsythe variance test [ N = 7 (WT), 7 (Mertk KO), 6 (Mertk CR3 dKO)]. (G) Whole mount retina images to show vehicle treated Axl KO mice with RGCs (Brn3a+; red) expressing nuclear p-cJUN (white) and microglia present (IBA1+; green). Scale bar = 500 μm. Boxed regions show enlarged area (G′) with yellow arrows indicating double positive p-cJUN and Brn3a staining. Scale bars = 100-10 μm. (H) Whole mount retina images to show Axl KO PLX treated mice with depletion of microglia (IBA1; green) and RGCs (Brn3a+; red) with nuclear p-cJUN (white) expression remaining. Boxed regions show enlarged area (H′) with yellow arrows indicating double positive p-cJUN and Brn3a staining. Scale bars = 100 to 10 μm.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_5010/pmc12715010/pmc12715010__fncel-19-1719791-g002.jpg)