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Cell growth and proliferation are increased in the K-Ras revertant A549 cells. (A) 2D colony-formation assay performed with the A549 NT , A549 REV1 , and A549 REV2 cells as described in Materials and Methods . Representative images of three independent experiments are shown. Data on the graph represent the average number of colonies of nine replicates from three independent experiments ± SD ( n = 9). (B) 3D soft agar tumorgenicity assay performed as described in Materials and Methods . Representative images of three independent experiments are shown. Data on the graph represent the measured diameter of every colony in a field of view from three independent experiments ± SD ( n = 27−37 colonies). (C) MTT assay comparing cell growth in the different strains, performed as described in Materials and Methods . Data on the graph represent measurements from four independent experiments normalized to the control A549 cells ± SD ( n = 4). (D) MTT assay performed with cells pretreated with 10 μM rapamycin or 0.1% DMSO as control (Ctrl). Data on the graph represent measurements from three independent experiments normalized to the A549 NT DMSO control ± SD ( n = 3). (E) MTT assay performed on cells treated with NT, K-Ras, or Pan-Ras siRNAs. Data on the graph represent measurements from three independent experiments normalized to the control NT <t>siRNA</t> condition ± SD ( n = 3). Adjusted p values: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. ns, nonstatistically significant.

Signalsilence Rictor Sirna, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rictor+8649s/SignalSilence+Rictor+siRNA+I/pmc10848948-111-33-39
Average 93 stars, based on 9 article reviews

signalsilence rictor sirna - by Bioz Stars, 2026-09

93/100 stars

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1) Product Images from "CRISPR-mediated reversion of oncogenic KRAS mutation results in increased proliferation and reveals independent roles of Ras and mTORC2 in the migration of A549 lung cancer cells"

Article Title: CRISPR-mediated reversion of oncogenic KRAS mutation results in increased proliferation and reveals independent roles of Ras and mTORC2 in the migration of A549 lung cancer cells

Journal: Molecular Biology of the Cell

doi: 10.1091/mbc.E23-05-0152

Figure Legend Snippet: Cell growth and proliferation are increased in the K-Ras revertant A549 cells. (A) 2D colony-formation assay performed with the A549 NT , A549 REV1 , and A549 REV2 cells as described in Materials and Methods . Representative images of three independent experiments are shown. Data on the graph represent the average number of colonies of nine replicates from three independent experiments ± SD ( n = 9). (B) 3D soft agar tumorgenicity assay performed as described in Materials and Methods . Representative images of three independent experiments are shown. Data on the graph represent the measured diameter of every colony in a field of view from three independent experiments ± SD ( n = 27−37 colonies). (C) MTT assay comparing cell growth in the different strains, performed as described in Materials and Methods . Data on the graph represent measurements from four independent experiments normalized to the control A549 cells ± SD ( n = 4). (D) MTT assay performed with cells pretreated with 10 μM rapamycin or 0.1% DMSO as control (Ctrl). Data on the graph represent measurements from three independent experiments normalized to the A549 NT DMSO control ± SD ( n = 3). (E) MTT assay performed on cells treated with NT, K-Ras, or Pan-Ras siRNAs. Data on the graph represent measurements from three independent experiments normalized to the control NT siRNA condition ± SD ( n = 3). Adjusted p values: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. ns, nonstatistically significant.

Techniques Used: Colony Assay, MTT Assay, Control

K-Ras promotes A549 cell migration independently of its mutation. (A) Wound closure cell-migration assays were performed with A549 NT , A549 REV1 , and A549 REV2 cells as described in Materials and Methods . Representative images of three independent experiments are shown, taken immediately after wounding (0 h) and after 24 h. Data on the graph represent measured wound closure (migration distances) in five areas of each wound from three independent experiments ± SD ( n = 15). (B) Wound-healing migration assays were performed with A549 NT , A549 REV1 , and A549 REV2 subjected to siRNA-mediated knockdown of K-Ras alone, of K-, H-, and N-Ras (Pan-Ras siRNA) or treated with NT siRNA control. The data were analyzed and graphed as described in (A). Adjusted p values for the differences between siRNA-mediated knockdowns and the NT siRNA control for each strain: ## p < 0.01; ### p < 0.001; #### p < 0.0001. ns, nonstatistically significant.

Figure Legend Snippet: K-Ras promotes A549 cell migration independently of its mutation. (A) Wound closure cell-migration assays were performed with A549 NT , A549 REV1 , and A549 REV2 cells as described in Materials and Methods . Representative images of three independent experiments are shown, taken immediately after wounding (0 h) and after 24 h. Data on the graph represent measured wound closure (migration distances) in five areas of each wound from three independent experiments ± SD ( n = 15). (B) Wound-healing migration assays were performed with A549 NT , A549 REV1 , and A549 REV2 subjected to siRNA-mediated knockdown of K-Ras alone, of K-, H-, and N-Ras (Pan-Ras siRNA) or treated with NT siRNA control. The data were analyzed and graphed as described in (A). Adjusted p values for the differences between siRNA-mediated knockdowns and the NT siRNA control for each strain: ## p < 0.01; ### p < 0.001; #### p < 0.0001. ns, nonstatistically significant.

Techniques Used: Migration, Mutagenesis, Knockdown, Control

mTORC2 mediates the migration of A549 cells independently of K-Ras’s mutational status. (A and B) Wound closure migration assays were performed with the A549 NT , A549 REV1 , and A549 REV2 cells treated with 10 μM rapamycin (mTORC1 inhibitor), 10 nM PP242 (mTORC1/mTORC2 inhibitor), or 0.1% DMSO control (A), or subjected to siRNA-mediated Rictor knockdown or treated with NT siRNA control (B), as described in Materials and Methods . Rictor immunoblots were performed to verify its siRNA-mediated knockdown and the data shown are representative of three independent experiments. Data on graphs represent measured migration distances in five areas of each wound from three independent experiments ± SD ( n = 15). (C) Transwell 3D invasion assays were performed with the cells and conditions as described in (A). Representative images of three independent experiments are shown. Data on graph represent the average number of cells that have invaded per area measured from three separate experiments ± SD ( n = 3). **** adjusted p value < 0.0001. Adjusted p values for the differences between treatment conditions and their respective control for each strain: #, p < 0.05; ##, p < 0.01; ###, p < 0.001; ####, p < 0.0001. ns, nonstatistically significant.

Figure Legend Snippet: mTORC2 mediates the migration of A549 cells independently of K-Ras’s mutational status. (A and B) Wound closure migration assays were performed with the A549 NT , A549 REV1 , and A549 REV2 cells treated with 10 μM rapamycin (mTORC1 inhibitor), 10 nM PP242 (mTORC1/mTORC2 inhibitor), or 0.1% DMSO control (A), or subjected to siRNA-mediated Rictor knockdown or treated with NT siRNA control (B), as described in Materials and Methods . Rictor immunoblots were performed to verify its siRNA-mediated knockdown and the data shown are representative of three independent experiments. Data on graphs represent measured migration distances in five areas of each wound from three independent experiments ± SD ( n = 15). (C) Transwell 3D invasion assays were performed with the cells and conditions as described in (A). Representative images of three independent experiments are shown. Data on graph represent the average number of cells that have invaded per area measured from three separate experiments ± SD ( n = 3). **** adjusted p value < 0.0001. Adjusted p values for the differences between treatment conditions and their respective control for each strain: #, p < 0.05; ##, p < 0.01; ###, p < 0.001; ####, p < 0.0001. ns, nonstatistically significant.

Techniques Used: Migration, Control, Knockdown, Western Blot

EGF stimulates mTORC2 activity independently of Ras in A549 cells. pAKT(S473) and pERK were assessed in cells pretreated with K-Ras siRNA, Pan-Ras siRNAs, or NT siRNA control, and stimulated or not with 100 ng/mL EGF as described in Materials and Methods . Ras knockdowns were verified by immunoblot. Representative immunoblots of three independent experiments are shown. Bands were quantified by densitometry and the data on the graph represent the ratio of pAKT(S473)/AKT expressed as percentage of A549 NT control stimulation measured from three separate experiments ± SD ( n = 3). Adjusted p value: **, p < 0.01. ns, nonstatistically significant.

Figure Legend Snippet: EGF stimulates mTORC2 activity independently of Ras in A549 cells. pAKT(S473) and pERK were assessed in cells pretreated with K-Ras siRNA, Pan-Ras siRNAs, or NT siRNA control, and stimulated or not with 100 ng/mL EGF as described in Materials and Methods . Ras knockdowns were verified by immunoblot. Representative immunoblots of three independent experiments are shown. Bands were quantified by densitometry and the data on the graph represent the ratio of pAKT(S473)/AKT expressed as percentage of A549 NT control stimulation measured from three separate experiments ± SD ( n = 3). Adjusted p value: **, p < 0.01. ns, nonstatistically significant.

Techniques Used: Activity Assay, Control, Western Blot

Small Interfering RNA:

Article Title: Rictor maintains endothelial integrity under shear stress
Article Snippet: A primary antibody against GAPDH (60,004–1) was purchased from Proteintech. .. Small interfering RNA (siRNA) targeting Rictor (8649s) was purchased from Cell Signaling Technology. .. Actin-Tracker Red-Rhodamine (C2207s) was obtained from Beyotime.

Article Title: Rictor maintains endothelial integrity under shear stress.
Article Snippet: A primary antibody against GAPDH (60,004–1) was purchased from Proteintech. .. Small interfering RNA (siRNA) targeting Rictor (8649s) was purchased from Cell Signaling Technology. .. ActinTracker Red-Rhodamine (C2207s) was obtained from Beyotime.

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Journal: Molecular Biology of the Cell

Article Title: CRISPR-mediated reversion of oncogenic KRAS mutation results in increased proliferation and reveals independent roles of Ras and mTORC2 in the migration of A549 lung cancer cells

doi: 10.1091/mbc.E23-05-0152

Figure Lengend Snippet: Cell growth and proliferation are increased in the K-Ras revertant A549 cells. (A) 2D colony-formation assay performed with the A549 NT , A549 REV1 , and A549 REV2 cells as described in Materials and Methods . Representative images of three independent experiments are shown. Data on the graph represent the average number of colonies of nine replicates from three independent experiments ± SD ( n = 9). (B) 3D soft agar tumorgenicity assay performed as described in Materials and Methods . Representative images of three independent experiments are shown. Data on the graph represent the measured diameter of every colony in a field of view from three independent experiments ± SD ( n = 27−37 colonies). (C) MTT assay comparing cell growth in the different strains, performed as described in Materials and Methods . Data on the graph represent measurements from four independent experiments normalized to the control A549 cells ± SD ( n = 4). (D) MTT assay performed with cells pretreated with 10 μM rapamycin or 0.1% DMSO as control (Ctrl). Data on the graph represent measurements from three independent experiments normalized to the A549 NT DMSO control ± SD ( n = 3). (E) MTT assay performed on cells treated with NT, K-Ras, or Pan-Ras siRNAs. Data on the graph represent measurements from three independent experiments normalized to the control NT siRNA condition ± SD ( n = 3). Adjusted p values: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. ns, nonstatistically significant.

Article Snippet: Dharmacon ON-TARGET plus Non-targeting siRNA Control Pool and SMARTPool of Human H-Ras (3265), Human N-Ras (4893), and Human K-Ras (3845) were obtained from Horizon Discovery (Waterbeach, UK), and SignalSilence Control siRNA (unconjugated) and SignalSilence Rictor siRNA were purchased from Cell Signaling Technology (Danvers, MA).

Techniques: Colony Assay, MTT Assay, Control

Journal: Molecular Biology of the Cell

Article Title: CRISPR-mediated reversion of oncogenic KRAS mutation results in increased proliferation and reveals independent roles of Ras and mTORC2 in the migration of A549 lung cancer cells

doi: 10.1091/mbc.E23-05-0152

Figure Lengend Snippet: K-Ras promotes A549 cell migration independently of its mutation. (A) Wound closure cell-migration assays were performed with A549 NT , A549 REV1 , and A549 REV2 cells as described in Materials and Methods . Representative images of three independent experiments are shown, taken immediately after wounding (0 h) and after 24 h. Data on the graph represent measured wound closure (migration distances) in five areas of each wound from three independent experiments ± SD ( n = 15). (B) Wound-healing migration assays were performed with A549 NT , A549 REV1 , and A549 REV2 subjected to siRNA-mediated knockdown of K-Ras alone, of K-, H-, and N-Ras (Pan-Ras siRNA) or treated with NT siRNA control. The data were analyzed and graphed as described in (A). Adjusted p values for the differences between siRNA-mediated knockdowns and the NT siRNA control for each strain: ## p < 0.01; ### p < 0.001; #### p < 0.0001. ns, nonstatistically significant.

Techniques: Migration, Mutagenesis, Knockdown, Control

Journal: Molecular Biology of the Cell

Article Title: CRISPR-mediated reversion of oncogenic KRAS mutation results in increased proliferation and reveals independent roles of Ras and mTORC2 in the migration of A549 lung cancer cells

doi: 10.1091/mbc.E23-05-0152

Figure Lengend Snippet: mTORC2 mediates the migration of A549 cells independently of K-Ras’s mutational status. (A and B) Wound closure migration assays were performed with the A549 NT , A549 REV1 , and A549 REV2 cells treated with 10 μM rapamycin (mTORC1 inhibitor), 10 nM PP242 (mTORC1/mTORC2 inhibitor), or 0.1% DMSO control (A), or subjected to siRNA-mediated Rictor knockdown or treated with NT siRNA control (B), as described in Materials and Methods . Rictor immunoblots were performed to verify its siRNA-mediated knockdown and the data shown are representative of three independent experiments. Data on graphs represent measured migration distances in five areas of each wound from three independent experiments ± SD ( n = 15). (C) Transwell 3D invasion assays were performed with the cells and conditions as described in (A). Representative images of three independent experiments are shown. Data on graph represent the average number of cells that have invaded per area measured from three separate experiments ± SD ( n = 3). **** adjusted p value < 0.0001. Adjusted p values for the differences between treatment conditions and their respective control for each strain: #, p < 0.05; ##, p < 0.01; ###, p < 0.001; ####, p < 0.0001. ns, nonstatistically significant.

Techniques: Migration, Control, Knockdown, Western Blot

Journal: Molecular Biology of the Cell

Article Title: CRISPR-mediated reversion of oncogenic KRAS mutation results in increased proliferation and reveals independent roles of Ras and mTORC2 in the migration of A549 lung cancer cells

doi: 10.1091/mbc.E23-05-0152

Figure Lengend Snippet: EGF stimulates mTORC2 activity independently of Ras in A549 cells. pAKT(S473) and pERK were assessed in cells pretreated with K-Ras siRNA, Pan-Ras siRNAs, or NT siRNA control, and stimulated or not with 100 ng/mL EGF as described in Materials and Methods . Ras knockdowns were verified by immunoblot. Representative immunoblots of three independent experiments are shown. Bands were quantified by densitometry and the data on the graph represent the ratio of pAKT(S473)/AKT expressed as percentage of A549 NT control stimulation measured from three separate experiments ± SD ( n = 3). Adjusted p value: **, p < 0.01. ns, nonstatistically significant.

Techniques: Activity Assay, Control, Western Blot

Cerebral ischemia up-regulated the expression of Sirt1/autophagy signaling in autopsied brains of stroke patients. Immunoblotting was performed on an equal amount of total protein sample (75 μg ) from healthy controls (n = 3) and stroke patients (n = 6). Respective total proteins and beta-actin were served as a loading control (Lc3-II/I), while Sirt1, Beclin-1, Atg-3, Atg-5, Atg-7, and Atg-12-5 were normalized to beta-actin (A–G). Graphs show changes in the expression of Sirt1/autophagy signaling. The densitometry values are expressed as mean ± SD (Raw immunoblot images were included in this article as supplementary Data).

Journal: Heliyon

Article Title: Up-regulation of Sirtuin-1/autophagy signaling in human cerebral ischemia: possible role in caspase-3 mediated apoptosis

doi: 10.1016/j.heliyon.2022.e12278

Figure Lengend Snippet: Cerebral ischemia up-regulated the expression of Sirt1/autophagy signaling in autopsied brains of stroke patients. Immunoblotting was performed on an equal amount of total protein sample (75 μg ) from healthy controls (n = 3) and stroke patients (n = 6). Respective total proteins and beta-actin were served as a loading control (Lc3-II/I), while Sirt1, Beclin-1, Atg-3, Atg-5, Atg-7, and Atg-12-5 were normalized to beta-actin (A–G). Graphs show changes in the expression of Sirt1/autophagy signaling. The densitometry values are expressed as mean ± SD (Raw immunoblot images were included in this article as supplementary Data).

Article Snippet: Sirt1 (CST#8649), Autophagy Antibody Sampler Kit (CST#4445), beta-actin (CST#8457), Anti-rabbit Ig-G-Alexa fluor-488 (CST#4412), Anti-mouse Ig-G Alexaflour-555 (CST# 4409), Prolong Gold Antifade Reagent with DAPI (CST#8961).

Techniques: Expressing, Western Blot, Control

Subcellular localization of Sirt1, Beclin-1, Atg-7, and Lc3-I/II in autopsied brains of stroke patients. Brain tissues from healthy subjects (n = 3) and stroke patients (n = 4) were paraffin-embedded and sectioned into 5–10 μm thick slices. (A–D) Graphical representation of changes in Immunoreactivity of Sirt1, Beclin-1, Atg-7, and Lc3-I/II from stroke patients and healthy subjects. The black arrow represents Sirt1, Beclin-1, Atg-7, and Lc3-I/II immunoreactivity for the respective group. Immunoreactivity was depicted in brown (DAB). The neuronal nucleus is counterstained with hematoxylin (magnification 400X; Scale Bar – 20 μm). (E) Triple immunofluorescence labeling for Lc3-I/II (green), Sirt1 (red), and DAPI (blue) in control (n = 3) and stroke human brains (n = 4). The representative image shows increased number of Lc3-I/II positive cells co-localized with Sirt1 in stroke brain compared to control (White arrows; magnification 630X; Scale Bar – 20 μm).

Journal: Heliyon

Article Title: Up-regulation of Sirtuin-1/autophagy signaling in human cerebral ischemia: possible role in caspase-3 mediated apoptosis

doi: 10.1016/j.heliyon.2022.e12278

Figure Lengend Snippet: Subcellular localization of Sirt1, Beclin-1, Atg-7, and Lc3-I/II in autopsied brains of stroke patients. Brain tissues from healthy subjects (n = 3) and stroke patients (n = 4) were paraffin-embedded and sectioned into 5–10 μm thick slices. (A–D) Graphical representation of changes in Immunoreactivity of Sirt1, Beclin-1, Atg-7, and Lc3-I/II from stroke patients and healthy subjects. The black arrow represents Sirt1, Beclin-1, Atg-7, and Lc3-I/II immunoreactivity for the respective group. Immunoreactivity was depicted in brown (DAB). The neuronal nucleus is counterstained with hematoxylin (magnification 400X; Scale Bar – 20 μm). (E) Triple immunofluorescence labeling for Lc3-I/II (green), Sirt1 (red), and DAPI (blue) in control (n = 3) and stroke human brains (n = 4). The representative image shows increased number of Lc3-I/II positive cells co-localized with Sirt1 in stroke brain compared to control (White arrows; magnification 630X; Scale Bar – 20 μm).

Techniques: Immunofluorescence, Labeling, Control

SIRT1 protein expression in the five different experimental groups. Data are presented as mean ± SEM (n=5 rats/group). **P<0.01 vs. controls. ##P<0.01 vs. HFD. ++P<0.01 vs. HFD-Q. SIRT1, sirtuin 1; HFD, high fat diabetic; Q, quercetin; CC, compound-C.

Journal: Experimental and Therapeutic Medicine

Article Title: Quercetin modulates AMPK/SIRT1/NF-κB signaling to inhibit inflammatory/oxidative stress responses in diabetic high fat diet-induced atherosclerosis in the rat carotid artery

doi: 10.3892/etm.2020.9410

Figure Lengend Snippet: SIRT1 protein expression in the five different experimental groups. Data are presented as mean ± SEM (n=5 rats/group). **P<0.01 vs. controls. ##P<0.01 vs. HFD. ++P<0.01 vs. HFD-Q. SIRT1, sirtuin 1; HFD, high fat diabetic; Q, quercetin; CC, compound-C.

Article Snippet: Following blocking with 5% skimmed milk for 1 h at room temperature, the membranes were incubated with primary antibodies against SIRT1 (cat. no. 8649S; 1:1,500; Cell Signaling Technology, Inc.) and β-actin (cat. no. 3700S; 1:5,000; Cell Signaling Technology, Inc.) overnight at 4 ̊C.

Techniques: Expressing

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1) Product Images from "CRISPR-mediated reversion of oncogenic KRAS mutation results in increased proliferation and reveals independent roles of Ras and mTORC2 in the migration of A549 lung cancer cells"

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