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Proteintech nti mlc2
Nti Mlc2, supplied by Proteintech, used in various techniques. Bioz Stars score: 94/100, based on 205 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Related Articles

Synthesized:

Article Title: RHOA lactylation at oncogenic hotspots promotes oncogenic activity and protein stabilization.
Article Snippet: Human KCTD13 and LDHA gene were amplified from MDA-MB231 cDNA library and subcloned into pLVX-Puro vector. pCMV-3×FLAG-USP9X(human)-Neo (P56773) and pCMV-USP33(human)-3×Myc-Neo(P71806) were obtained from MiaoLingBio, China. .. The DNA fragments of Mb-Pyl Klacr-RS/Pyl-tRNA pair were synthesized by Beijing Tsingke Biotech Co., Ltd. Antibodies against RHOA and MLC2 were purchased from Proteintech (catalog no. 10749-1-AP and 10906-1-AP). .. Antibody against FLAG was purchased from Sigma-Aldrich (catalog no. F3165).

Article Title: RHOA lactylation at oncogenic hotspots promotes oncogenic activity and protein stabilization
Article Snippet: Human KCTD13 and LDHA gene were amplified from MDA-MB-231 cDNA library and subcloned into pLVX-Puro vector. pCMV-3 × FLAG-USP9X(human)-Neo ( P56773 ) and pCMV-USP33(human)−3 × Myc-Neo( P71806 ) were obtained from MiaoLingBio, China. .. The DNA fragments of Mb-Pyl Klacr-RS/Pyl-tRNA pair were synthesized by Beijing Tsingke Biotech Co., Ltd. Antibodies against RHOA and MLC2 were purchased from Proteintech (catalog no. 10749–1-AP and 10,906–1-AP). .. Antibody against FLAG was purchased from Sigma-Aldrich (catalog no. F3165).

Membrane:

Article Title: CapG promoted nasopharyngeal carcinoma cell motility involving Rho motility pathway independent of ROCK
Article Snippet: Denatured protein was separated via 10% SDS polyacrylamide gel electrophoresis and transferred to PVDF membrane (0.22 μm, Merck Millipore, MA, USA). .. Membrane was blocked with TBST solution (20 mM Tris, 137 mM NaCl, 0.1% Triton X-100, pH 7.6 ± 0.1) containing 5% nonfat milk for 1 h at room temperature, followed by incubation with specific primary antibodies at 4 °C for 16 h. The primary antibodies included alpha tubulin (1:2000; ab52866, Abcam, Cambridge, UK), CapG (1:1000; ab155688, Abcam, Cambridge, UK), MLC2 (1:2000; 10906-1-AP, Proteintech, IL, USA), and phospho-MLC2 (p-MLC2) (1:1000; no. 3674, Cell Signaling Technology, MA, USA). ..

Article Title: CapG promoted nasopharyngeal carcinoma cell motility involving Rho motility pathway independent of ROCK.
Article Snippet: Denatured protein was separated via 10% SDS polyacrylamide gel electrophoresis and transferred to PVDF membrane (0.22 μm, Merck Millipore, MA, USA). .. Membrane was blocked with TBST solution (20 mM Tris, 137 mM NaCl, 0.1% Triton X-100, pH 7.6 ± 0.1) containing 5% nonfat milk for 1 h at room temperature, followed by incubation with specific primary antibodies at 4 °C for 16 h. The primary antibodies included alpha tubulin (1:2000; ab52866, Abcam, Cambridge, UK), CapG (1:1000; ab155688, Abcam, Cambridge, UK), MLC2 (1:2000; 10906-1-AP, Proteintech, IL, USA), and phospho-MLC2 (p-MLC2) (1:1000; no. 3674, Cell Signaling Technology, MA, USA). ..

Incubation:

Article Title: CapG promoted nasopharyngeal carcinoma cell motility involving Rho motility pathway independent of ROCK
Article Snippet: Denatured protein was separated via 10% SDS polyacrylamide gel electrophoresis and transferred to PVDF membrane (0.22 μm, Merck Millipore, MA, USA). .. Membrane was blocked with TBST solution (20 mM Tris, 137 mM NaCl, 0.1% Triton X-100, pH 7.6 ± 0.1) containing 5% nonfat milk for 1 h at room temperature, followed by incubation with specific primary antibodies at 4 °C for 16 h. The primary antibodies included alpha tubulin (1:2000; ab52866, Abcam, Cambridge, UK), CapG (1:1000; ab155688, Abcam, Cambridge, UK), MLC2 (1:2000; 10906-1-AP, Proteintech, IL, USA), and phospho-MLC2 (p-MLC2) (1:1000; no. 3674, Cell Signaling Technology, MA, USA). ..

Article Title: CapG promoted nasopharyngeal carcinoma cell motility involving Rho motility pathway independent of ROCK.
Article Snippet: Denatured protein was separated via 10% SDS polyacrylamide gel electrophoresis and transferred to PVDF membrane (0.22 μm, Merck Millipore, MA, USA). .. Membrane was blocked with TBST solution (20 mM Tris, 137 mM NaCl, 0.1% Triton X-100, pH 7.6 ± 0.1) containing 5% nonfat milk for 1 h at room temperature, followed by incubation with specific primary antibodies at 4 °C for 16 h. The primary antibodies included alpha tubulin (1:2000; ab52866, Abcam, Cambridge, UK), CapG (1:1000; ab155688, Abcam, Cambridge, UK), MLC2 (1:2000; 10906-1-AP, Proteintech, IL, USA), and phospho-MLC2 (p-MLC2) (1:1000; no. 3674, Cell Signaling Technology, MA, USA). ..

Article Title: The Hippo pathway links adipocyte plasticity to adipose tissue fibrosis
Article Snippet: .. Membranes were blotted with primary antibodies against HSP90 (Santa Cruz, sc-13119, 1:10,000), α-Tubulin (Sigma, T6199, 1:10,000), Lamin B1 (Proteintech, 66095-1-Ig, 1:10,000), LATS1 (Cell Signaling, 3477, 1:1000), LATS2 (Bethyl Laboratories, A300-479A, 1:1000), YAP (Cell Signaling, 4912 or 14074, 1:1000), TAZ (Cell Signaling, 4883 or 83669, 1:1000), p-YAP (Ser 112) (Cell Signaling, 4911), p-TAZ (Ser 89) (Cell Signaling, 59971, 1:1000), HSL (Cell Signaling, 4107, 1:1000), Perilipin 1 (Vala Sciences, 4854, 1:10,000), SMAD2/3 (Cell Signaling, 8685, 1:1000), p-SMAD2 (Ser465/467)/SMAD3 (Ser423/425) (Cell Signaling, 8828, 1:1000), αSMA (Cell Signaling, 19245, 1:5000), FLAG (Abmart, M20008, 1:5000), Caspase 3 (Cell Signaling, 9662,1:1000), MST1 (Cell Signaling, 3682, 1:1000), MST2 (Cell Signaling, 3952, 1:1000), p-SAPK/JNK (Thr183/Tyr185) (Cell Signaling, 4668, 1:1000), SAPK/JNK (Cell Signaling, 9252, 1:1000), p-AKT (Ser473) (Cell Signaling, 4060, 1:1000), AKT (Cell Signaling, 9272, 1:1000), p-p38 MAPK (Thr180/Tyr182) (Cell Signaling, 4511, 1:1000), p38 MAPK (Cell Signaling, 8690, 1:1000), p-MLC2 (Ser19) (Cell Signaling, 3671, 1:1000), MLC2 (Proteintech, 10906-1-AP, 1:1000), ERK1/ERK2 (ABclonal, A10613, 1:1000), p-ERK1(T202/Y204)/ERK2(T185/Y187) (ABclonal, AP0472, 1:1000), GFP (Thermo Scientific, A-11120, 1:1000), Myc (Cell Signaling, 2276, 1:2000) and Col1a1 (Cell Signaling, 72026, 1:1000) followed by incubation with HRP-conjugated anti-rabbit secondary antibody (Thermo Scientific, 31460, 1:10,000) or anti-mouse secondary antibody (Thermo Scientific, 32430, 1:10,000). .. Nuclear and cytoplasmic fractionation were performed using the Nuclear and Cytoplasmic Protein Extraction Kit (Beyotime, P0028), according to the manufacturer’s manual.



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(A) Schematic overview of ROCK1 construct transfection used to assess entosis induction. (B) Representative immunoblots showing ROCK1, phosphorylated <t>MLC2</t> (pMLC2), and tubulin (loading control) in MCF7 cells transfected with GFP, GFP–ROCK1, or constitutively active GFP–ROCK1 Δ3. Immunoblot analysis of pMLC2 was performed 6 h post-transfection, whereas ROCK1 expression was assessed at 24 h. (C) Representative 3D confocal images of live MCF7 cells expressing GFP–ROCK1 Δ3 and stained with SiR-Actin (red) and Hoechst (blue). White arrows indicate entotic structures. Boxed regions showing representative entotic structures are magnified, with corresponding orthogonal z-stack views shown alongside each image. (D) Quantification of entotic events in MCF7 cells expressing GFP–ROCK1 or GFP–ROCK1 Δ3. (E) Distribution percentages of GFP-positive cells participating in CIC structures as outer cells, inner cells, or both. (F) Representative time-lapse imaging (phase contrast, Hoechst, and GFP) capturing an entotic event in MCF7 cells expressing GFP–ROCK1 Δ3. Yellow dotted lines outline inner cells, and red dotted lines outline host cells. Experiments were performed in biological triplicate (n = 3). Data are presented as mean ± SEM. Statistical analysis in panel D was performed using a paired two-tailed Student’s t-test (**p < 0.01), whereas panel E was analyzed using two-way ANOVA followed by Bonferroni post hoc test (*p < 0.05, **p < 0.01, ****p < 0.0001).
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(A) Schematic overview of ROCK1 construct transfection used to assess entosis induction. (B) Representative immunoblots showing ROCK1, phosphorylated <t>MLC2</t> (pMLC2), and tubulin (loading control) in MCF7 cells transfected with GFP, GFP–ROCK1, or constitutively active GFP–ROCK1 Δ3. Immunoblot analysis of pMLC2 was performed 6 h post-transfection, whereas ROCK1 expression was assessed at 24 h. (C) Representative 3D confocal images of live MCF7 cells expressing GFP–ROCK1 Δ3 and stained with SiR-Actin (red) and Hoechst (blue). White arrows indicate entotic structures. Boxed regions showing representative entotic structures are magnified, with corresponding orthogonal z-stack views shown alongside each image. (D) Quantification of entotic events in MCF7 cells expressing GFP–ROCK1 or GFP–ROCK1 Δ3. (E) Distribution percentages of GFP-positive cells participating in CIC structures as outer cells, inner cells, or both. (F) Representative time-lapse imaging (phase contrast, Hoechst, and GFP) capturing an entotic event in MCF7 cells expressing GFP–ROCK1 Δ3. Yellow dotted lines outline inner cells, and red dotted lines outline host cells. Experiments were performed in biological triplicate (n = 3). Data are presented as mean ± SEM. Statistical analysis in panel D was performed using a paired two-tailed Student’s t-test (**p < 0.01), whereas panel E was analyzed using two-way ANOVA followed by Bonferroni post hoc test (*p < 0.05, **p < 0.01, ****p < 0.0001).
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(A) Schematic overview of ROCK1 construct transfection used to assess entosis induction. (B) Representative immunoblots showing ROCK1, phosphorylated <t>MLC2</t> (pMLC2), and tubulin (loading control) in MCF7 cells transfected with GFP, GFP–ROCK1, or constitutively active GFP–ROCK1 Δ3. Immunoblot analysis of pMLC2 was performed 6 h post-transfection, whereas ROCK1 expression was assessed at 24 h. (C) Representative 3D confocal images of live MCF7 cells expressing GFP–ROCK1 Δ3 and stained with SiR-Actin (red) and Hoechst (blue). White arrows indicate entotic structures. Boxed regions showing representative entotic structures are magnified, with corresponding orthogonal z-stack views shown alongside each image. (D) Quantification of entotic events in MCF7 cells expressing GFP–ROCK1 or GFP–ROCK1 Δ3. (E) Distribution percentages of GFP-positive cells participating in CIC structures as outer cells, inner cells, or both. (F) Representative time-lapse imaging (phase contrast, Hoechst, and GFP) capturing an entotic event in MCF7 cells expressing GFP–ROCK1 Δ3. Yellow dotted lines outline inner cells, and red dotted lines outline host cells. Experiments were performed in biological triplicate (n = 3). Data are presented as mean ± SEM. Statistical analysis in panel D was performed using a paired two-tailed Student’s t-test (**p < 0.01), whereas panel E was analyzed using two-way ANOVA followed by Bonferroni post hoc test (*p < 0.05, **p < 0.01, ****p < 0.0001).
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(A) Schematic overview of ROCK1 construct transfection used to assess entosis induction. (B) Representative immunoblots showing ROCK1, phosphorylated <t>MLC2</t> (pMLC2), and tubulin (loading control) in MCF7 cells transfected with GFP, GFP–ROCK1, or constitutively active GFP–ROCK1 Δ3. Immunoblot analysis of pMLC2 was performed 6 h post-transfection, whereas ROCK1 expression was assessed at 24 h. (C) Representative 3D confocal images of live MCF7 cells expressing GFP–ROCK1 Δ3 and stained with SiR-Actin (red) and Hoechst (blue). White arrows indicate entotic structures. Boxed regions showing representative entotic structures are magnified, with corresponding orthogonal z-stack views shown alongside each image. (D) Quantification of entotic events in MCF7 cells expressing GFP–ROCK1 or GFP–ROCK1 Δ3. (E) Distribution percentages of GFP-positive cells participating in CIC structures as outer cells, inner cells, or both. (F) Representative time-lapse imaging (phase contrast, Hoechst, and GFP) capturing an entotic event in MCF7 cells expressing GFP–ROCK1 Δ3. Yellow dotted lines outline inner cells, and red dotted lines outline host cells. Experiments were performed in biological triplicate (n = 3). Data are presented as mean ± SEM. Statistical analysis in panel D was performed using a paired two-tailed Student’s t-test (**p < 0.01), whereas panel E was analyzed using two-way ANOVA followed by Bonferroni post hoc test (*p < 0.05, **p < 0.01, ****p < 0.0001).
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Image Search Results


(A) Schematic overview of ROCK1 construct transfection used to assess entosis induction. (B) Representative immunoblots showing ROCK1, phosphorylated MLC2 (pMLC2), and tubulin (loading control) in MCF7 cells transfected with GFP, GFP–ROCK1, or constitutively active GFP–ROCK1 Δ3. Immunoblot analysis of pMLC2 was performed 6 h post-transfection, whereas ROCK1 expression was assessed at 24 h. (C) Representative 3D confocal images of live MCF7 cells expressing GFP–ROCK1 Δ3 and stained with SiR-Actin (red) and Hoechst (blue). White arrows indicate entotic structures. Boxed regions showing representative entotic structures are magnified, with corresponding orthogonal z-stack views shown alongside each image. (D) Quantification of entotic events in MCF7 cells expressing GFP–ROCK1 or GFP–ROCK1 Δ3. (E) Distribution percentages of GFP-positive cells participating in CIC structures as outer cells, inner cells, or both. (F) Representative time-lapse imaging (phase contrast, Hoechst, and GFP) capturing an entotic event in MCF7 cells expressing GFP–ROCK1 Δ3. Yellow dotted lines outline inner cells, and red dotted lines outline host cells. Experiments were performed in biological triplicate (n = 3). Data are presented as mean ± SEM. Statistical analysis in panel D was performed using a paired two-tailed Student’s t-test (**p < 0.01), whereas panel E was analyzed using two-way ANOVA followed by Bonferroni post hoc test (*p < 0.05, **p < 0.01, ****p < 0.0001).

Journal: bioRxiv

Article Title: Plastin-3 membrane recruitment drives cell-in-cell invasion during entosis

doi: 10.64898/2026.03.17.709257

Figure Lengend Snippet: (A) Schematic overview of ROCK1 construct transfection used to assess entosis induction. (B) Representative immunoblots showing ROCK1, phosphorylated MLC2 (pMLC2), and tubulin (loading control) in MCF7 cells transfected with GFP, GFP–ROCK1, or constitutively active GFP–ROCK1 Δ3. Immunoblot analysis of pMLC2 was performed 6 h post-transfection, whereas ROCK1 expression was assessed at 24 h. (C) Representative 3D confocal images of live MCF7 cells expressing GFP–ROCK1 Δ3 and stained with SiR-Actin (red) and Hoechst (blue). White arrows indicate entotic structures. Boxed regions showing representative entotic structures are magnified, with corresponding orthogonal z-stack views shown alongside each image. (D) Quantification of entotic events in MCF7 cells expressing GFP–ROCK1 or GFP–ROCK1 Δ3. (E) Distribution percentages of GFP-positive cells participating in CIC structures as outer cells, inner cells, or both. (F) Representative time-lapse imaging (phase contrast, Hoechst, and GFP) capturing an entotic event in MCF7 cells expressing GFP–ROCK1 Δ3. Yellow dotted lines outline inner cells, and red dotted lines outline host cells. Experiments were performed in biological triplicate (n = 3). Data are presented as mean ± SEM. Statistical analysis in panel D was performed using a paired two-tailed Student’s t-test (**p < 0.01), whereas panel E was analyzed using two-way ANOVA followed by Bonferroni post hoc test (*p < 0.05, **p < 0.01, ****p < 0.0001).

Article Snippet: Membranes were blocked in 5% non-fat milk in TBS-T or in 5% BSA in TBS-T when detecting phospho-specific antibodies and incubated with primary antibodies against ROCK1 (rabbit monoclonal, 1:1000; Cell Signaling Technology, #28999S), RhoA (rabbit monoclonal, 1:1000; Cell Signaling Technology, #2117S), PLS2 (rabbit monoclonal, 1:1000; Atlas Antibodies, #HPA019493), PLS3 (mouse monoclonal, 1:1000; Invitrogen, #MA5-27772), α-tubulin (rabbit polyclonal, 1:3000; Cell Signaling Technology, #2144S), phospho-MLC2 (Ser19) (rabbit, 1:1000; Cell Signaling Technology, #3671S), and GFP (chicken polyclonal, 1:1000; Rockland Immunochemicals, #600-901-215).

Techniques: Construct, Transfection, Western Blot, Control, Expressing, Staining, Imaging, Two Tailed Test