Journal: Carbohydrate polymers

Article Title: Stability and bioactivity of chitosan as a transfection agent in primary human cell cultures: A case for chitosan-only controls.

doi: 10.1016/j.carbpol.2017.10.021

Figure Lengend Snippet: Fig. 1. Plasmid DNA containing human CD59 on a high-yield promoter (pDNA, A) was incorporated into chitosan microparticles (pCsM, B). Both pCsM and microparticles without pDNA (CsM, C) were precipitated from chitosan polymers (Cs, D). All chitosan treatments were prepared and delivered in mildly acidic Vehicle (pH 5.6, E).

Article Snippet: TrueClone cDNA plasmids containing human CD59 (Origene, Rockville, Maryland) were expanded in Alpha-Gold competent Escherichia coli bacteria (Origene).

Techniques: Plasmid Preparation

Journal: Carbohydrate polymers

Article Title: Stability and bioactivity of chitosan as a transfection agent in primary human cell cultures: A case for chitosan-only controls.

doi: 10.1016/j.carbpol.2017.10.021

Figure Lengend Snippet: Fig. 4. Mean fluorescent intensity from flow cytometry was used to approximate average CD59 expression vSMC (A). CD59 is transiently depressed by pCsM, CsM and Cs (B, p ≤0.001) but recovers to baseline by 24 h despite the presence of Vehicle (C). In the absence of chitosan, Vehicle decreases CD59 expression.

Article Snippet: TrueClone cDNA plasmids containing human CD59 (Origene, Rockville, Maryland) were expanded in Alpha-Gold competent Escherichia coli bacteria (Origene).

Techniques: Cytometry, Expressing

Journal: Carbohydrate polymers

Article Title: Stability and bioactivity of chitosan as a transfection agent in primary human cell cultures: A case for chitosan-only controls.

doi: 10.1016/j.carbpol.2017.10.021

Figure Lengend Snippet: Fig. 5. Flow cytometry gates to determine Live CD59-positive vSMC were set based on living and EtOH-killed controls (A). Chitosan (pCsM, CsM, Cs) initially decreased the number of CD59-positive cells (B, p ≤0.001), an effect that was rapidly, potently and durably reversed, especially in the case of Cs (C, *p ≤0.01, §p ≤0.05).

Article Snippet: TrueClone cDNA plasmids containing human CD59 (Origene, Rockville, Maryland) were expanded in Alpha-Gold competent Escherichia coli bacteria (Origene).

Techniques: Flow Cytometry

Journal: Carbohydrate polymers

Article Title: Stability and bioactivity of chitosan as a transfection agent in primary human cell cultures: A case for chitosan-only controls.

doi: 10.1016/j.carbpol.2017.10.021

Figure Lengend Snippet: Fig. 6. CD59 mRNA was elevated after treatment with pDNA (p < 0.05) and pCsM (p < 0.01) but down-regulated by treatment with Vehicle (p < 0.05).

Article Snippet: TrueClone cDNA plasmids containing human CD59 (Origene, Rockville, Maryland) were expanded in Alpha-Gold competent Escherichia coli bacteria (Origene).

Techniques:

Journal: Acta pharmacologica Sinica

Article Title: Ginsenoside Rb1 alleviates colitis in mice via activation of endoplasmic reticulum-resident E3 ubiquitin ligase Hrd1 signaling pathway.

doi: 10.1038/s41401-020-00561-9

Figure Lengend Snippet: Fig. 2 Effects of the ginsenoside Rb1 (GRb1) on inflammation and apoptosis in a DSS-induced mouse model of colitis. Expression levels of the colonic cytokines a MPO, b IL-1β, c IL-6, and d TNF-α were determined by ELISA. Western blot analysis of the inflammation-related proteins e iNOS, f COX2, and g p-NF-κB-p65 (calculated as p-NF-κB-p65/NF-κB-p65) and the apoptosis-related protein h cleaved-caspase 3 (cl-caspase 3), calculated as cl-caspase 3/caspase 3. Data are expressed as the mean ± SD. Values in the sham group were set to 100%, and other values are given relative to those in the sham group, **P < 0.01 compared with the sham group; ##P < 0.01 compared with the DSS control group; n = 3 samples for Western blot experiments; n = 6 samples for other experiments.

Article Snippet: Antibodies against Hrd1(13473-1-AP), CHOP(15204-1-AP), Fas (13098-1-AP), caspase 8(13423-1-AP), caspase 3 (19677-1-AP), cleaved-caspase 3 (cl-caspase 3) (19677-1-AP), and iNOS (14142-1-AP) were purchased from Proteintech Co., Ltd. (Wuhan, China).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Western Blot, Control

Journal: Acta pharmacologica Sinica

Article Title: Ginsenoside Rb1 alleviates colitis in mice via activation of endoplasmic reticulum-resident E3 ubiquitin ligase Hrd1 signaling pathway.

doi: 10.1038/s41401-020-00561-9

Figure Lengend Snippet: Fig. 3 Effects of the ginsenoside Rb1 (GRb1) on ER stress and Hrd1 in a DSS-induced mouse model of colitis. Western blot analysis of the ER stress marker proteins a GRP78, b PERK, c CHOP, and d caspase 12. e Western blot analysis of Hrd1. f Expression of Hrd1 determined by real- time PCR. g Immunohistochemical staining of Hrd1 in the colonic epithelium. Scale bars, 200, 100, and 50 μm. Data are expressed as the mean ± SD; **P < 0.01 compared with the sham group; ##P < 0.01 compared with the DSS control group; n = 3 samples for Western blot experiments; n = 6 samples for other experiments.

Article Snippet: Antibodies against Hrd1(13473-1-AP), CHOP(15204-1-AP), Fas (13098-1-AP), caspase 8(13423-1-AP), caspase 3 (19677-1-AP), cleaved-caspase 3 (cl-caspase 3) (19677-1-AP), and iNOS (14142-1-AP) were purchased from Proteintech Co., Ltd. (Wuhan, China).

Techniques: Western Blot, Marker, Expressing, Real-time Polymerase Chain Reaction, Immunohistochemical staining, Staining, Control

Journal: Acta pharmacologica Sinica

Article Title: Ginsenoside Rb1 alleviates colitis in mice via activation of endoplasmic reticulum-resident E3 ubiquitin ligase Hrd1 signaling pathway.

doi: 10.1038/s41401-020-00561-9

Figure Lengend Snippet: Fig. 5 Effect of the ginsenoside Rb1 (GRb1) on apoptosis. a Effect of GRb1 (200 μM) on IEC-6 cell apoptosis, as determined by flow cytometry. Effect of GRb1 on the expression of b Fas and c caspase 8 in vivo. Effect of GRb1 (200 μM) on the expression of Fas in LPS-treated IEC-6 cells in the absence (d) and presence (e) of siRNA targeting Hrd1. f Effects of GRb1 (200 μM) on IEC-6 cell apoptosis in the presence of siRNA targeting Hrd1. Data are expressed as the mean ± SD. **P < 0.01 compared with the corresponding control group; ##P < 0.01 compared with the DSS control group. n = 3 samples for Western blot experiments, n = 6 samples for other experiments.

Article Snippet: Antibodies against Hrd1(13473-1-AP), CHOP(15204-1-AP), Fas (13098-1-AP), caspase 8(13423-1-AP), caspase 3 (19677-1-AP), cleaved-caspase 3 (cl-caspase 3) (19677-1-AP), and iNOS (14142-1-AP) were purchased from Proteintech Co., Ltd. (Wuhan, China).

Techniques: Cytometry, Expressing, In Vivo, Control, Western Blot

Journal: Acta pharmacologica Sinica

Article Title: Ginsenoside Rb1 alleviates colitis in mice via activation of endoplasmic reticulum-resident E3 ubiquitin ligase Hrd1 signaling pathway.

doi: 10.1038/s41401-020-00561-9

Figure Lengend Snippet: Fig. 7 Effects of the ginsenoside Rb1 (GRb1) on Hrd1, ER stress, apoptosis, and inflammation in a TNBS-induced colitis model. Western blot analysis of a Hrd1, b CHOP, c GRP78, d cleaved-caspase 3 (cl-caspase 3) (calculated as cl-caspase 3/caspase 3), and e p-NF-κB-p65 (calculated as p-NF-κB-p65/NF-κB-p65). Data are expressed as the mean ± SD. **P < 0.01 compared with the sham group (n = 3 mice); ##P < 0.01 compared with the TNBS-induced colitis group; n = 3 mice.

Article Snippet: Antibodies against Hrd1(13473-1-AP), CHOP(15204-1-AP), Fas (13098-1-AP), caspase 8(13423-1-AP), caspase 3 (19677-1-AP), cleaved-caspase 3 (cl-caspase 3) (19677-1-AP), and iNOS (14142-1-AP) were purchased from Proteintech Co., Ltd. (Wuhan, China).

Techniques: Western Blot

Journal: Science advances

Article Title: Codependencies of mTORC1 signaling and endolysosomal actin structures.

doi: 10.1126/sciadv.add9084

Figure Lengend Snippet: Fig. 4. RAGC dissociation from endolysosomes upon disruption of the branched actin network. (A) Cells in complete medium were treated with vehicle (DMSO) or with CK-666 for 60 min. After fixation, cells were stained for F-actin, CD63, and LAMTOR4. (B) Quantification of LAMTOR4-positive vesicles/cell in cells with the indicated treatment. (C and D) MDA-MB-231 cells were transfected with vectors expressing GFP-tagged LAMTOR1 (C) or FLAG-tagged LAMTOR4 (D) or corresponding empty vectors. Cells in complete medium were treated with vehicle (DMSO) or CK-666 for 60 min and lysed, and proteins were immunoprecipitated using the GFP- or FLAG-Trap procedure, respectively. Bound partners were detected by immunoblotting with the indicated antibodies. (E) MDA-MB-231 cells were stained for F-actin, RAGC, and CD63. The nucleus contour is shown with a dashed line. Scale bars, 10 and 2 μm (insets). (F) Quantification of RAGC-positive vesicles/cell in cells with the indicated treatment. (G to I) MDA-MB-231 cells transiently expressing FLAG-tagged RAGCQ120L or RAGCS75N or transfected with control empty FLAG vector (Ctrl) were fixed and labeled for FLAG tag and p4E-BP1 (see fig. S5, A and B) or mTOR. Quantification of the mean intensity of p4E-BP1 staining in the indicated cell population normalized to the mean intensity in control cells in (G). Number of mTOR-positive vesicular structures per cell in (H). Median and quartiles shown in the violin plots. Intensity of mTOR signal is shown using a fire lookup table in (I). Asterisk denotes FLAG-positive cell. Scale bar, 10 μm (I). All numerical data and statistical tests are provided in table S5.

Article Snippet: Plasmid FLAGHA-pcDNA3.1- was a gift from A. Antebi (Addgene plasmid #52535), plasmids pRK5 Flag-RAGC(S75N) (Addgene plasmid #99724), pRK5 Flag RAGC(Q120L) (Addgene plasmid #99725), N1-p18-EGFP (Addgene plasmid #42334, GFP-LAMTOR1), and pRK5-FLAG-C7orf59 (Addgene plasmid #42332, FLAGLAMTOR4) were provided by D. Sabatini (7, 57).

Techniques: Disruption, Staining, Transfection, Expressing, Immunoprecipitation, Western Blot, Control, Plasmid Preparation, Labeling, FLAG-tag

Journal: Journal of Cellular and Molecular Medicine

Article Title: AIMP3 inhibits cell growth and metastasis of lung adenocarcinoma through activating a miR‐96‐5p‐AIMP3‐p53 axis

doi: 10.1111/jcmm.16344

Figure Lengend Snippet: AIMP3 expression in clinical non‐small cell lung tumour and adjacent tissues. A, Representative IHC staining of AIMP3 on NSCLC and adjacent tissues. B, AIMP3 expression plotted based on immunohistochemical score. C, Seven random selected human NSCLC tumours and their paired adjacent normal tissues were tested using AIMP3 specific antibody. A, adjacent normal tissues; T, tumours. D, Density analysis of Western blot results. * P < .05 compared with adjacent normal tissues

Article Snippet: The proteins were detected by the following primary antibodies: AIMP3 (Proteintech, 10805‐1‐AP), p53 (Proteintech, 10442‐1‐AP), N‐cadherin(Cell Signaling, 13116), E‐cadherin (Cell Signaling, 3195), Vimentin (Cell Signaling, 5741), Snail (Cell Signaling, 3879), Slug (Cell Signaling, 9585), ATM (Proteintech, 27156‐1‐AP), p‐ATM (Abcam, ab81292), β‐actin (Cell Signaling, 3700) and GAPDH (Proteintech, 60004‐1‐Ig) and followed by incubation with a secondary antibody.

Techniques: Expressing, Immunohistochemistry, Immunohistochemical staining, Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: AIMP3 inhibits cell growth and metastasis of lung adenocarcinoma through activating a miR‐96‐5p‐AIMP3‐p53 axis

doi: 10.1111/jcmm.16344

Figure Lengend Snippet: Overexpression of AIMP3 suppresses the proliferation of NSCLC cells. A, The expression of AIMP3 protein in six NSCLC cell lines indicated and HBE cells. Relative fold changes were determined by densitometry and normalized to GAPDH. B, H1299 and A549 cells were transiently transfected with different amounts of HA‐AIMP3 for 24 h. Cell lysates were extracted and used to detect the protein levels of AIMP3 and p53. C, Saturation density assay. Cells transfected with indicated amount of AIMP3 were cultured in full medium for 6 days, trypsinized and counted. Data are presented as mean ± SEM (n = 3). D, Low serum assay. Cells transiently transfected with AIMP3 were cultured in 1% FBS RPMI1640 medium. At the indicated times, cells were trypsinized and counted. Data are presented as mean ± SEM (n = 3). E, Colony formation assay. 500 transiently transfected cells were seeded in 6‐well plates in 5% FBS RPMI1640 medium, respectively. After 14 days, cells were fixed and stained with 2% crystal violet. F, The number of colonies was counted and data are presented as mean ± SEM (n = 3). * P < .05, ** P < .01, *** P < .001 by Student's t test

Article Snippet: The proteins were detected by the following primary antibodies: AIMP3 (Proteintech, 10805‐1‐AP), p53 (Proteintech, 10442‐1‐AP), N‐cadherin(Cell Signaling, 13116), E‐cadherin (Cell Signaling, 3195), Vimentin (Cell Signaling, 5741), Snail (Cell Signaling, 3879), Slug (Cell Signaling, 9585), ATM (Proteintech, 27156‐1‐AP), p‐ATM (Abcam, ab81292), β‐actin (Cell Signaling, 3700) and GAPDH (Proteintech, 60004‐1‐Ig) and followed by incubation with a secondary antibody.

Techniques: Over Expression, Expressing, Transfection, Cell Culture, Serum Assay, Colony Assay, Staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: AIMP3 inhibits cell growth and metastasis of lung adenocarcinoma through activating a miR‐96‐5p‐AIMP3‐p53 axis

doi: 10.1111/jcmm.16344

Figure Lengend Snippet: AIMP3 suppresses the migration of NSCLC cells. A, Transwell migration assay. 10 4 transiently transfected H1299 and A549 cells were cultured in 1% FBS RPMI1640 medium and seeded in transwell chambers. 10 h later, cells were fixed, stained and photographed. B, The crystal violet was dissolved in DMSO, and the absorbance was tested. Data are presented as mean ± SEM (n = 3). C, Cells were transiently transfected with AIMP3 or control vector. The expression of N‐cadherin, E‐cadherin, Vimentin, Snail and Slug was analysed by Western blot. D, Relative fold changes were determined by densitometry and normalized to β‐actin

Article Snippet: The proteins were detected by the following primary antibodies: AIMP3 (Proteintech, 10805‐1‐AP), p53 (Proteintech, 10442‐1‐AP), N‐cadherin(Cell Signaling, 13116), E‐cadherin (Cell Signaling, 3195), Vimentin (Cell Signaling, 5741), Snail (Cell Signaling, 3879), Slug (Cell Signaling, 9585), ATM (Proteintech, 27156‐1‐AP), p‐ATM (Abcam, ab81292), β‐actin (Cell Signaling, 3700) and GAPDH (Proteintech, 60004‐1‐Ig) and followed by incubation with a secondary antibody.

Techniques: Migration, Transwell Migration Assay, Transfection, Cell Culture, Staining, Control, Plasmid Preparation, Expressing, Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: AIMP3 inhibits cell growth and metastasis of lung adenocarcinoma through activating a miR‐96‐5p‐AIMP3‐p53 axis

doi: 10.1111/jcmm.16344

Figure Lengend Snippet: AIMP3 induces cell cycle arrest at G2/M phase and apoptosis depends on p53. A, Cell cycle analysis of A549 cells transfected with different amounts of AIMP3 for 48 h. Cells were collected and analysed by flow cytometry after propidium iodide staining. The inserts showed the proportion of cells for each phase and were marked with different colours (violet: G0/G1 phase, green: S phase and gold: G2/M phase). B, The quantification of cell number in each phase of cell cycle. C, Examination of apoptotic cells in A549 cells transfected with indicated amount of AIMP3. D, Quantitative analysis of the percentage of apoptotic cells was shown in Figure C. Data are presented as mean ± SEM (n = 3); * P < .05, ** P < .01. E, The effect of AIMP3 on p53‐dependent transcription of NOXA , PUMA and p21 . A549 cells were transiently transfected with indicated amount of AIMP3 and incubated for 24 h. Total RNAs were extracted and used for RT‐PCR detection. F, A549 cells were co‐transfected with p21 promoter‐reporter vector and different amounts of AIMP3 expression plasmid (0, 50, 100 ng). Relative luciferase activity values are presented as mean ± SEM (n = 3); ** P < .01 by Student's t test. G, A549 cells were transiently transfected with HA‐AIMP3 for 24 h, and whole cell lysates were immunoprecipitated with HA antibody (mouse) or mouse IgG as control. The precipitates were immunoblotted with ATM antibody. H, Cells were transiently transfected with AIMP3 or control vector. The expression of indicated proteins was analysed by Western blot

Article Snippet: The proteins were detected by the following primary antibodies: AIMP3 (Proteintech, 10805‐1‐AP), p53 (Proteintech, 10442‐1‐AP), N‐cadherin(Cell Signaling, 13116), E‐cadherin (Cell Signaling, 3195), Vimentin (Cell Signaling, 5741), Snail (Cell Signaling, 3879), Slug (Cell Signaling, 9585), ATM (Proteintech, 27156‐1‐AP), p‐ATM (Abcam, ab81292), β‐actin (Cell Signaling, 3700) and GAPDH (Proteintech, 60004‐1‐Ig) and followed by incubation with a secondary antibody.

Techniques: Cell Cycle Assay, Transfection, Flow Cytometry, Staining, Incubation, Reverse Transcription Polymerase Chain Reaction, Plasmid Preparation, Expressing, Luciferase, Activity Assay, Immunoprecipitation, Control, Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: AIMP3 inhibits cell growth and metastasis of lung adenocarcinoma through activating a miR‐96‐5p‐AIMP3‐p53 axis

doi: 10.1111/jcmm.16344

Figure Lengend Snippet: AIMP3 suppresses tumour growth in mouse xenograft model. A, A549 cells stably expressing empty vector or AIMP3 were subcutaneously injected into nude mice , and the expression of AIMP3 was confirmed by western blot. Tumour volume was measured every 3 days and is shown as the mean ± SEM (empty vector, Mock: n = 8, AIMP3: n = 8). Significant differences were determined by one‐way ANOVA, followed by Dunnett's test; * P < .05 (versus empty). A representative image of tumour xenografts harvested at day 30. B, Tumour weights from the two groups are represented. C, BALB/c nude mice (n = 8 per group) received a tail vein injection of A549 cells stably expressing empty vector or AIMP3. The number of metastatic lung nodules in individual mice was counted under a dissection microscope. Data represent the mean ± SEM * P < .05 vs empty vector

Article Snippet: The proteins were detected by the following primary antibodies: AIMP3 (Proteintech, 10805‐1‐AP), p53 (Proteintech, 10442‐1‐AP), N‐cadherin(Cell Signaling, 13116), E‐cadherin (Cell Signaling, 3195), Vimentin (Cell Signaling, 5741), Snail (Cell Signaling, 3879), Slug (Cell Signaling, 9585), ATM (Proteintech, 27156‐1‐AP), p‐ATM (Abcam, ab81292), β‐actin (Cell Signaling, 3700) and GAPDH (Proteintech, 60004‐1‐Ig) and followed by incubation with a secondary antibody.

Techniques: Stable Transfection, Expressing, Plasmid Preparation, Injection, Western Blot, Dissection, Microscopy

Journal: Journal of Cellular and Molecular Medicine

Article Title: AIMP3 inhibits cell growth and metastasis of lung adenocarcinoma through activating a miR‐96‐5p‐AIMP3‐p53 axis

doi: 10.1111/jcmm.16344

Figure Lengend Snippet: AIMP3 is a direct target of miR‐96‐5p. A, Prediction of miR‐96‐5p binding sites in 3’UTR of AIMP3 using Target Scan Human 7.2 software. B, A549 cells were co‐transfected with AIMP3 3’‐UTR plasmid and miR‐96‐5p mimic (left panel) or miR‐96‐5p inhibitor (right panel). Relative luciferase activity values are presented as mean ± SEM (n = 3); * P < .05, ** P < .01. C, The expression of miR‐96‐5p in six NSCLC cell lines and HBE cells. U6 was used as an internal control. * P < .05, ** P < .01, *** P < .001. D, miR‐96‐5p expression in 36 pairs of NSCLC and matched adjacent normal tissues. *** P < .001. E, Correlation between AIMP3 and miR‐96‐5p expression in 36 NSCLC tissues. F, Up‐regulation of miR‐96‐5p suppressed the expression of AIMP3 protein and mRNA in A549 cells. G, Inhibition of miR‐96‐5p increased both the mRNA and protein levels of AIMP3

Article Snippet: The proteins were detected by the following primary antibodies: AIMP3 (Proteintech, 10805‐1‐AP), p53 (Proteintech, 10442‐1‐AP), N‐cadherin(Cell Signaling, 13116), E‐cadherin (Cell Signaling, 3195), Vimentin (Cell Signaling, 5741), Snail (Cell Signaling, 3879), Slug (Cell Signaling, 9585), ATM (Proteintech, 27156‐1‐AP), p‐ATM (Abcam, ab81292), β‐actin (Cell Signaling, 3700) and GAPDH (Proteintech, 60004‐1‐Ig) and followed by incubation with a secondary antibody.

Techniques: Binding Assay, Software, Transfection, Plasmid Preparation, Luciferase, Activity Assay, Expressing, Control, Inhibition

Journal: Journal of Cellular and Molecular Medicine

Article Title: AIMP3 inhibits cell growth and metastasis of lung adenocarcinoma through activating a miR‐96‐5p‐AIMP3‐p53 axis

doi: 10.1111/jcmm.16344

Figure Lengend Snippet: The effect of miR‐96‐5p on the growth of NSLSC cells. A, Saturation density assay. H1299 and A549 cells transiently transfected with miRNA control, miR‐96‐5p mimic or inhibitor were cultured in full medium. 6 days later, cells were trypsinized and counted. Data are presented as mean ± SEM (n = 3); * P < .05, ** P < .01 by Student's t test. B, Low serum assay. H1299 and A549 cells transiently transfected with miRNA control, miR‐96‐5p mimic were cultured in 1% FBS RPMI1640 medium. Cells were trypsinized and counted at indicated times. Data are presented as mean ± SEM (n = 3); ** P < .01 by Student's t test. C, Colony formation assay. Five hundred transiently transfected H1299 and A549 cells were seeded in 6‐well plates in 5% FBS RPMI1640 medium, respectively. After 14 days, cells were fixed and stained. The number of colonies was counted and data are presented as mean ± SEM (n = 3); ** P < .01 by Student's t test. D, Transwell migration assay. 10 4 transiently transfected H1299 and A549 cells were cultured in 1% FBS RPMI1640 medium and seeded in transwell chambers. 10 h later, cells were fixed, stained and photographed. Then, the crystal violet was dissolved in DMSO and the absorbance was tested. Data are presented as mean ± SEM (n = 3); ** P < .01 by Student's t test. E, qPCR was used to examine the miR‐96‐5p level in transfected A549 cells. F, Western blot was performed to detect the expression of AIMP3 and p53 in these cells. G, Tumour weights of tumours from control or miR‐96‐5p mimic transfected A549 xenografts at sacrifice. n = 8 for each group. H, The number of metastatic lung nodules in individual mice received a tail vein injection of A549 cells transfected control or miR‐96‐5p mimic was counted. Data represent the mean ± SEM* P < .05 vs control

Article Snippet: The proteins were detected by the following primary antibodies: AIMP3 (Proteintech, 10805‐1‐AP), p53 (Proteintech, 10442‐1‐AP), N‐cadherin(Cell Signaling, 13116), E‐cadherin (Cell Signaling, 3195), Vimentin (Cell Signaling, 5741), Snail (Cell Signaling, 3879), Slug (Cell Signaling, 9585), ATM (Proteintech, 27156‐1‐AP), p‐ATM (Abcam, ab81292), β‐actin (Cell Signaling, 3700) and GAPDH (Proteintech, 60004‐1‐Ig) and followed by incubation with a secondary antibody.

Techniques: Transfection, Control, Cell Culture, Serum Assay, Colony Assay, Staining, Transwell Migration Assay, Western Blot, Expressing, Injection