Journal: Oncotarget

Article Title: Asiatic acid protects against hepatic ischemia/reperfusion injury by inactivation of Kupffer cells via PPARγ/NLRP3 inflammasome signaling pathway

doi: 10.18632/oncotarget.21151

Figure Lengend Snippet: Male C57BL/6 mice were pretreated with either AA (30 mg/kg) or vehicle 1 h before hepatic I/R surgery. After 6 h reperfusion, liver tissues and serum samples were harvested. (A) Representative histological staining (H&E) of ischemic liver tissue. Results representative of 4-6 mice/group. Liver damage, evaluated by Suzuki’s histological score. Scale bar: 30μm. (B) Hepatocellular function in serum samples was evaluated by sALT and sAST levels (IU/L). Results expressed as mean±SD (n=4-6 samples/group). (C) & (D) Liver neutrophils and monocytes/macrophages were detected by immunofluorescent staining. Quantification of Ly6G+ cells and CD11b+ cells per HPF. Representative of 4-6 mice/group. Scale bar: 30μm. (E) RT-qPCR for detection of IL-6, TNF-α, and CXCL1 in ischemic livers (n=4-6 samples/group). * Significant difference ( P < 0.05) compared with corresponding control. # Significant difference ( P < 0.05) compared with I/R.

Article Snippet: Alexa Fluor 488-conjugated anti-Ly6G, Alexa Fluor 488-conjugated anti-F4/80 and Alexa Fluor 594-conjugated anti-CD11b were purchased from Bioss, China.

Techniques: Staining, Quantitative RT-PCR

Journal: Oncotarget

Article Title: Asiatic acid protects against hepatic ischemia/reperfusion injury by inactivation of Kupffer cells via PPARγ/NLRP3 inflammasome signaling pathway

doi: 10.18632/oncotarget.21151

Figure Lengend Snippet: Mice were pretreated with GW9662 (2 mg/kg, i.p.) or vehicle 30 min prior to treatment with AA, followed by an I/R insult. Liver tissues and serum samples were harvested 6 h after reperfusion. (A) Representative histological staining, Ly6G+ cells and CD11b+ cells infiltration and immunohistochemical staining of NLRP3 in ischemic livers. Scale bar: 30μm. (B) Suzuki’s histological score and sALT. (C) Quantitative analysis of infiltrated Ly6G+ cells and CD11b+ cells. (D) mRNA levels of IL-6, TNF-α and CXCL1, as well as mRNA levels of NLRP3 and IL-1β were determined using RT-qPCR. (E) Protein expressions of NLRP3, ASC, pro-caspase-1, cleaved caspase-1 p10, pro-IL-1β, IL-1β, PPARγ, Bcl-2, pro-caspase-3 and caspase-3 were detected using Western blot analysis. (F) Quantitative analysis of NLRP3 positively-stained cells. (G) ELISA analysis of IL-1β levels in animal serums. (H) Co-immunoprecipitation and Western blot for assessing the NLRP3 inflammasome assembly. The results are presented as the mean±SD of 4-6 animals per group. Blots shown are representative of 3 experiments with similar results. * P < 0.05 compared between the indicated groups. ** P < 0.01 compared between the indicated groups.

Article Snippet: Alexa Fluor 488-conjugated anti-Ly6G, Alexa Fluor 488-conjugated anti-F4/80 and Alexa Fluor 594-conjugated anti-CD11b were purchased from Bioss, China.

Techniques: Staining, Immunohistochemical staining, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Immunoprecipitation

Journal: Oncotarget

Article Title: Asiatic acid protects against hepatic ischemia/reperfusion injury by inactivation of Kupffer cells via PPARγ/NLRP3 inflammasome signaling pathway

doi: 10.18632/oncotarget.21151

Figure Lengend Snippet: Mice were pretreated with GdCl3 24 h prior to AA treatment or AA+GW9662 treatment, followed by an I/R insult. Liver tissues and serum samples were harvested after 6 h reperfusion. (A) Representative histological staining, Ly6G+ cells and CD11b+ cells infiltration and immunohistochemical staining of NLRP3 in ischemic livers. Scale bar: 30μm. (B) Suzuki’s histological score and sALT. (C) Quantitative analysis of infiltrated Ly6G+ cells and CD11b+ cells. (D) mRNA levels of IL-6, TNF-α and CXCL1, as well as mRNA levels of NLRP3 and IL-1β were determined using RT-qPCR. (E) Protein expressions of NLRP3, ASC, pro-caspase-1, cleaved caspase-1 p10, pro-IL-1β, IL-1β, PPARγ, Bcl-2, pro-caspase-3 and caspase-3 were detected using Western blot analysis. (F) Quantitative analysis of NLRP3 positively-stained cells. (G) ELISA analysis of IL-1β levels in animal serums. The results are presented as the mean±SD of 4-6 animals per group. Blots shown are representative of 3 experiments with similar results. * P < 0.05 compared between the indicated groups. ** P < 0.01 compared between the indicated groups.

Article Snippet: Alexa Fluor 488-conjugated anti-Ly6G, Alexa Fluor 488-conjugated anti-F4/80 and Alexa Fluor 594-conjugated anti-CD11b were purchased from Bioss, China.

Techniques: Staining, Immunohistochemical staining, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Journal of Inflammation Research

Article Title: The Role of Bone Morphogenetic Protein 4 in Microglial Polarization in the Process of Neuropathic Pain

doi: 10.2147/JIR.S356531

Figure Lengend Snippet: Exogenous BMP4-induced allodynia, microglial activation and polarization. ( A ) PWT in rats after intrathecal BMP4 application showed a significant effect on time: F (4.143, 66.29) = 16.09, P<0.01, left; F (3.960, 63.35) = 12.67, P<0.01, right, intrathecal application: F (1, 16) = 161.3, P<0.01, left; F (1, 16) = 160.5, P<0.01, right and interaction: F (7, 112) = 11.76, P<0.01, left; F (7, 112) = 5.653, P<0.01, right. Compared with the Sham group, rats in the BMP4 group developed a significant decrease in bilateral PWT for the whole first week, P<0.01, n=9 at each time point for both groups. ( B ) Representative Western blotting showed a sustained increase of CD11b (P<0.01) and CD16 (P<0.01) expressions for the whole 1st week in the BMP4 group compared with the Sham group. Meanwhile, ARG-1 levels increased at day 1 (P<0.01) and day 4 (P<0.01), then fell below the levels of the Sham group (P<0.01). n=3 for each column. ( C and D ) Double-immunofluorescence further detected that compared with the Sham group (P7), CD11b expression was elevated in the dorsal horn of spinal cord after BMP4 treatment. Moreover, expressions of CD16 and ARG-1 showed similar pattern with Western blotting results, which were both mainly accumulated with CD11b + cells. n=3 for each column. Two-way ANOVA, followed by a Bonferroni test ( A ) and one-way ANOVA, followed by Sidak’s multiple comparisons test ( B – D ) were performed to analyze the statistical differences. *Represented P<0.05 and **Represented P<0.01 compared with the Sham group.

Article Snippet: All sections were rinsed in 0.2%Triton X-100 mixed with 5% donkey serum for 40 min at room temperature and then incubated overnight with primary antibodies at 4°C at the following dilutions: mouse anti-CD11b (SM262PS, 1:50, OriGene, MD, US), rabbit anti-CD16 (1:50, Affinity) and rabbit anti-ARG1 (1:100, Affinity).

Techniques: Activation Assay, Western Blot, Immunofluorescence, Expressing

Journal: Journal of Inflammation Research

Article Title: The Role of Bone Morphogenetic Protein 4 in Microglial Polarization in the Process of Neuropathic Pain

doi: 10.2147/JIR.S356531

Figure Lengend Snippet: Exogenous BMP4 drove the imbalance of the M1/M2 ratio. ( A ) The steps and gating strategies of CD11b + microglia. ( B and C ) Representative flow cytometry images showed that exogenous BMP4 induces a sustained elevation of CD11b + CD86 + M1 microglia from day 1 to day 7. In the meantime, CD172 + CD11b + M2 microglia increased at day 1 and day 4, then fell to the Sham level at day 7. Moreover, the M1/M2 ratio persistently increased from P1 to P7 throughout the whole 1st week, indicating BMP4 induced a lasting imbalance of M1/M2 ratio. n=3 for each time point for both groups. One-way ANOVA, followed by Sidak’s multiple comparisons test were performed to analyze the statistical differences. *Represented P<0.05 and **Represented P<0.01 compared with the Sham group.

Article Snippet: All sections were rinsed in 0.2%Triton X-100 mixed with 5% donkey serum for 40 min at room temperature and then incubated overnight with primary antibodies at 4°C at the following dilutions: mouse anti-CD11b (SM262PS, 1:50, OriGene, MD, US), rabbit anti-CD16 (1:50, Affinity) and rabbit anti-ARG1 (1:100, Affinity).

Techniques: Flow Cytometry

Journal: Journal of Inflammation Research

Article Title: The Role of Bone Morphogenetic Protein 4 in Microglial Polarization in the Process of Neuropathic Pain

doi: 10.2147/JIR.S356531

Figure Lengend Snippet: Noggin relieved allodynia, attenuated microglial activation, and changed the polarized pattern after SNL. ( A ) PWT in rats receiving intrathecal Noggin application after SNL showed a significant effect on time: F (3.463, 55.41) = 162.3, P<0.01, intrathecal application: F (1, 16) = 44.85, P<0.01 and interaction: F (7, 112) = 3.540, P<0.01. Compared with the SNL group, Noggin application provided marked pain relief at day 3, day 4, day 5 and day 7 after SNL. n=9 at each time point for both groups. ( B and C ) Representative Western blotting showed SNL induced CD11b and CD16 upregulation at the first week, while ARG-1 expression remained unchanged compared with the Sham group; Moreover, Noggin effectively decreased the CD11b expressions at P1 and P4 and the CD16 expressions consecutively for the whole week. Simultaneously, the ARG-1 levels significantly elevated from P1 to P7 after Noggin treatment. n=3 for each column. Two-way ANOVA, followed by a Bonferroni test (A) and one-way ANOVA, followed by Sidak’s multiple comparisons test (B and C) were performed to analyze the statistical differences. **Represented P<0.01 compared with the Sham group, # Represented P<0.05 and ## Represented P<0.01 compared with the SNL group.

Article Snippet: All sections were rinsed in 0.2%Triton X-100 mixed with 5% donkey serum for 40 min at room temperature and then incubated overnight with primary antibodies at 4°C at the following dilutions: mouse anti-CD11b (SM262PS, 1:50, OriGene, MD, US), rabbit anti-CD16 (1:50, Affinity) and rabbit anti-ARG1 (1:100, Affinity).

Techniques: Activation Assay, Western Blot, Expressing

Journal: Journal of Inflammation Research

Article Title: The Role of Bone Morphogenetic Protein 4 in Microglial Polarization in the Process of Neuropathic Pain

doi: 10.2147/JIR.S356531

Figure Lengend Snippet: Noggin reversed the imbalance of the M1-M2 ratio in the process of NP. ( A and B ) Representative flow cytometry images showed that SNL induced a significant elevation of CD11b + CD86 + M1 microglia from day 1 to day 7, while CD172 + CD11b + M2 microglia decreased at P4 in the SNL group compared with the Sham group. Then, in the SNL+NOG group, Noggin treatment markedly decreased the proportion of CD11b+CD86+ M1 microglia from P1 to P7, while increased the proportion of CD172 + CD11b + M2 microglia at P7. Moreover, M1/M2 ratio markedly decreased from P4 to P7 after Noggin treatment. n=3 for each time point. One-way ANOVA, followed by Sidak’s multiple comparisons test were performed to analyze the statistical differences. *Represented P<0.05 and **Represented P<0.01 compared with the Sham group; # Represented P<0.05 and ## Represented P<0.01 compared with the SNL group.

Article Snippet: All sections were rinsed in 0.2%Triton X-100 mixed with 5% donkey serum for 40 min at room temperature and then incubated overnight with primary antibodies at 4°C at the following dilutions: mouse anti-CD11b (SM262PS, 1:50, OriGene, MD, US), rabbit anti-CD16 (1:50, Affinity) and rabbit anti-ARG1 (1:100, Affinity).

Techniques: Flow Cytometry

Journal: General Psychiatry

Article Title: Microglia activation in the offspring of prenatal Poly I: C exposed rats: a PET imaging and immunohistochemistry study

doi: 10.1136/gpsych-2018-000006

Figure Lengend Snippet: (A) CD11b-immunopositive cells in the control group (A, C) and thetreatment group (B, D) Group comparison of OD values in the prefrontal cortex and hippocampus

Article Snippet: Then sections were incubated in a humidified chamber with the rabbit primary antibody against CD11b (1:100; Boster Biological Engineering, China) in PBS overnight at 4°C.

Techniques: Control, Comparison

Journal: Journal of dairy science

Article Title: Direct labeling of milk cells without centrifugation for counting total and differential somatic cells using flow cytometry.

doi: 10.3168/jds.2022-22038

Figure Lengend Snippet: Figure 4. Cell differentiation using antibodies against integrin α-M (CD11b) and major histocompatibility complex class II (MHCII). Gating of total cells (P1) based on positive DNA staining in a dot plot with forward scatter height (FSC-H) on the x-axis and PB450-H on the y-axis (A, C, E). Dot plot for cell differentiation, selecting the events from gate P1 from the dot plot (A, C, E). The JF646-labeled MHCII antibodies are represented in the AP-H channel on the x-axis and the fluorescein isothiocyanate (FITC)-labeled CD11b antibodies on the FITC-H chan- nel are represented on the y-axis, respectively. The distinction of 2 cell populations, CD11b+/MHCII− events in the upper left gate (MHCII−) and CD11b+/MHCII+ events in the upper right gate (MHCII+; B, D, F) were performed. Representative quarter milk samples with low SCC (<10,000 cells/mL; A, B), high (>10,000 < 10,000,000 cells/mL; C, D), and very high (>10,000,000 cells/mL; E, F) are shown.

Article Snippet: The obtained cells were resuspended, and the cell suspension (20 μL) was pipetted onto the coated slides and incubated at RT for 2 h. The cells were fixed using 100 μL of PBS containing 4% paraformaldehyde (PBS-PFA 4%) at 4°C for 1 h. Nonspecific binding sites were saturated with PBSBSA 5% at RT for 1 h. Primary antibodies mouse antibovine CD11b (Clone MM10 A, WS0600B-100, Kingfisher Biotech) and mouse anti-bovine MHCII (Clone CC108, MCA5656) were diluted in a ratio of 1:200 in PBS-BSA 5% before overnight incubation at 4°C.

Techniques: Cell Differentiation, Immunopeptidomics, Staining, Labeling

Journal: Journal of dairy science

Article Title: Direct labeling of milk cells without centrifugation for counting total and differential somatic cells using flow cytometry.

doi: 10.3168/jds.2022-22038

Figure Lengend Snippet: Figure 5. Verification of the differential somatic cell count (dsCC) method by fluorescence microscopy. Immunofluorescence staining of polymorphonuclear cells (PMNC) cells isolated from bull blood (upper lane), bovine macrophages (Woo et al., 2006; middle lane), and raw milk (lower lane), incubated with integrin α-M (CD11b; secondary antibody FITC labeled, green; B, F, J) and major histocompatibility complex class II (MHCII; secondary antibody APC labeled, red; C, G, K) antibodies, together with the nuclear counterstained Hoechst (A, D, I) and the merged pictures of all 3 channels (panels D, H, L).

Article Snippet: The obtained cells were resuspended, and the cell suspension (20 μL) was pipetted onto the coated slides and incubated at RT for 2 h. The cells were fixed using 100 μL of PBS containing 4% paraformaldehyde (PBS-PFA 4%) at 4°C for 1 h. Nonspecific binding sites were saturated with PBSBSA 5% at RT for 1 h. Primary antibodies mouse antibovine CD11b (Clone MM10 A, WS0600B-100, Kingfisher Biotech) and mouse anti-bovine MHCII (Clone CC108, MCA5656) were diluted in a ratio of 1:200 in PBS-BSA 5% before overnight incubation at 4°C.

Techniques: Cell Counting, Fluorescence, Microscopy, Immunofluorescence, Staining, Isolation, Incubation, Labeling, Immunopeptidomics

Journal: Journal of Cellular and Molecular Medicine

Article Title: ATPR triggers acute myeloid leukaemia cells differentiation and cycle arrest via the RARα/LDHB/ERK‐glycolysis signalling axis

doi: 10.1111/jcmm.15353

Figure Lengend Snippet: ATPR inhibits cells proliferation and differentiation of AML cells. (A) AML cells were exposed to various concentrations (0, 10 −9 , 10 −8 , 10 −7 , 10 −6 , or 10 −5 M) of ATPR for 72 h, followed by the determination of cell viability using the CCK8 assay. (B) NB4 and MOLM‐13 cells were exposed to various concentrations (0, 10 −9 , 10 −8 , 10 −7 , 10 −6 , or 10 −5 M) of ATPR for 24–72 h, followed by the determination of cell viability using the CCK8 assay. (C) NB4 and MOLM‐13 cells were treated with ATPR (10 −6 M) for different durations (24–72 h). Immunofluorescence was used to detect KI67. (D) CFDA‐SE levels were assessed by flow cytometry. (E) NB4 and MOLM‐13 cells were treated with 0, 10 −9 , 10 −8 , 10 −7 , 10 −6 , or 10 −5 M) of ATPR for 24–72 h. The cell cycle distribution was analysed by flow cytometry (Mean ± SD, n = 3). (F) NB4 and MOLM‐13 cells were treated with ATPR (10 −6 M) for different durations (24–72 h). The protein expression of Cyclin A2, CDK4 and Cyclin D3 were assessed by Western blotting (Mean ± SD, n = 3). (G) NB4 and MOLM‐13 cells were stained with Wright‐Giemsa dye and the cell morphological features were observed under a microscopy. (H) CD11b expression was analysed by flow cytometer. (I) CD14 expression was analysed by flow cytometer. (J) NBT reduction experiment was performed to count the positive cell rate (Mean ± SD, n = 3). (K) The protein expression of PU.1 was determined by Western blotting (Mean ± SD, n = 3). * P < 0.05, ** P < 0.01 versus control group

Article Snippet: Human monoclonal LDHB, KI67 and CD11b antibody (Bioss, China) for IHC were used at a 1:300 dilution.

Techniques: CCK-8 Assay, Immunofluorescence, Flow Cytometry, Expressing, Western Blot, Staining, Microscopy

Journal: Journal of Cellular and Molecular Medicine

Article Title: ATPR triggers acute myeloid leukaemia cells differentiation and cycle arrest via the RARα/LDHB/ERK‐glycolysis signalling axis

doi: 10.1111/jcmm.15353

Figure Lengend Snippet: ATPR shows antileukaemic effects with RARα dependent. (A) The NB4 cells were treated with ATPR (10 −6 M) for different durations (24h–72 h). The protein expression of PML‐RARα, RARα, RARβ and RARγ was assessed by Western blotting (Mean ± SD, n = 3). (B) Values are presented as the mean ± SD (n = 3) of three in Quantitative real‐time PCR analysis of mRNA expression of RARα, RARβ, RARγ, CRABP2 and CYP26A1 treated with ATPR (10 −6 M) for 48h in NB4 cells‐dependent experiments. (C) The MOLM‐13 cells were treated with ATPR (10 −6 M) for different durations (24h–72h). The protein expression of RARα, RARβ and RARγ was assessed by Western blotting (Mean ± SD, n = 3). (D) Values are presented as the mean ± SD of three in Quantitative real‐time PCR analysis of mRNA expression of RARα, RARβ, RARγ, CRABP2 and CYP26A1 treated with ATPR (10 −6 M) for 48h in MOLM‐13 cells‐dependent experiments. (E) The cell cycle distribution was analysed by flow cytometry (Mean ± SD, n = 3). (F) CD11b expression was analysed by flow cytometer. (G) The protein expression of PU.1, Cyclin A2, CDK4 and Cyclin D3 was assessed by Western blotting (Mean ± SD, n = 3). (H) AML cells were exposed to various concentrations (0, 2, 4, 8 or 10mM) of 2‐DG for 72 h, followed by the determination of cell viability using the CCK8 assay. (I) AML cells were exposed to various concentrations (0, 10 −7 , 10 −6 , or 10 −5 M) of ATPR for 72 h, followed by the determination of the glycolysis rate using the glucose, lactic acid and ATP detection kit (Mean ± SD, n = 3). (J) Values are presented as the mean ± SD of three in Quantitative real‐time PCR analysis of mRNA expression of LDHB, LDHA, HK2, ENO1 and GAPDH treated with ATPR (10 −6 M) for 48h in MOLM‐13 and NB4 cells‐dependent experiments (K) The OCR was determined in MOLM‐13 and NB4 cells by Seahorse XF. * P < 0.05, ** P < 0.01 versus control group

Article Snippet: Human monoclonal LDHB, KI67 and CD11b antibody (Bioss, China) for IHC were used at a 1:300 dilution.

Techniques: Expressing, Western Blot, Real-time Polymerase Chain Reaction, Flow Cytometry, CCK-8 Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: ATPR triggers acute myeloid leukaemia cells differentiation and cycle arrest via the RARα/LDHB/ERK‐glycolysis signalling axis

doi: 10.1111/jcmm.15353

Figure Lengend Snippet: Knockdown LDHB expression inhibits proliferation while promoting differentiation and glycolysis of AML cells. (A) Western analysis of LDHB expression in AML patients PBMC compared with normal human PBMC (Mean ± SD, n = 3). (B) Western analysis of LDHB expression in AML cell lines (Mean ± SD, n = 3). (C, D) After treatment with ATPR (0, 10 −9 , 10 −8 , 10 −7 , 10 −6 , or 10 −5 M) for different durations (24–72 h). The protein expression of LDHB was assessed by Western blotting (Mean ± SD, n = 3). (E) NB4 and MOLM‐13 cells were treated with ATPR in the absence or in the presence of the RARα‐selective antagonist Ro 41‐5253. The protein expression of LDHB was assessed by Western blotting (Mean ± SD, n = 3). (F) Stable control and shLDHB‐transfected NB4 cells were observed by inversed fluorescent microscope. (G) The protein expression of LDHB was assessed by Western blotting after treatment with shLDHB for 7 days (Mean ± SD, n = 3). (H) Cell growth of NB4 and MPOLM‐13 cells after transfection with shLDHB as determined by the CCK‐8 assay at different time points (Mean ± SD, n = 3). (I) The cell cycle distribution was analysed by flow cytometry in NB4 and MOLM‐13 cells after LDHB depletion (Mean ± SD, n = 3). (J) Immunofluorescence was used to detect KI67 in NB4 and MOLM‐13 cells after LDHB depletion. (K) After LDHB depletion, NB4 and MOLM‐13 cells were stained with Wright‐Giemsa dye and the cell morphological features were observed under a microscopy. (L) NBT reduction experiment was performed to count the positive cell rate (Mean ± SD, n = 3). (M) CD11b and CD14 expression were analysed by flow cytometer in NB4 and MOLM‐13 cells after LDHB depletion (Mean ± SD, n = 3). (N) The protein expression of PU.1, Cyclin A2, CDK4 and Cyclin D3 was assessed by Western blotting after LDHB depletion (Mean ± SD, n = 3). * P < 0.05, ** P < 0.01 versus control group

Article Snippet: Human monoclonal LDHB, KI67 and CD11b antibody (Bioss, China) for IHC were used at a 1:300 dilution.

Techniques: Expressing, Western Blot, Transfection, Microscopy, CCK-8 Assay, Flow Cytometry, Immunofluorescence, Staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: ATPR triggers acute myeloid leukaemia cells differentiation and cycle arrest via the RARα/LDHB/ERK‐glycolysis signalling axis

doi: 10.1111/jcmm.15353

Figure Lengend Snippet: ATPR regulates the Raf/MEK/ERK signalling pathway through LDHB and the effects of LDHB on tumour growth in vivo (A) Protein expression of Raf, p‐MEK, MEK, p‐ERK and ERK was determined by Western blotting analysis after treatment with ATPR for different concentration (10 −7 ,10 −6 ,10 −5 ) (Mean ± SD, n = 3). (B) Protein expression of Raf, p‐MEK, MEK, p‐ERK and ERK was determined by western blotting analysis after treatment with ATPR in the absence or in the presence of the RARα‐selective antagonist Ro 41‐5253 (Mean ± SD, n = 3). (C) Protein expression of Raf, p‐MEK, MEK, p‐ERK and ERK was determined by Western blotting analysis after LDHB depletion in NB4 and MOLM‐13 cells (Mean ± SD, n = 3). (D) Representative two tumour tissues from vehicle control mice and ATPR‐treated mice group was fixed and immunohistochemistry staining for LDHB. (E, F) Tumour images and weights at experimental endpoints in NC and shLDHB xenografts (Mean ± SD, n = 6). (G) Tumour volumes were measured every day (Mean ± SD, n = 6). (H) Immunohistochemistry staining for LDHB, CD11b and KI67 of NC and shLDHB in NB4 and MOLM‐13 group. * P < 0.05, ** P < 0.01 versus control group

Article Snippet: Human monoclonal LDHB, KI67 and CD11b antibody (Bioss, China) for IHC were used at a 1:300 dilution.

Techniques: In Vivo, Expressing, Western Blot, Concentration Assay, Immunohistochemistry, Staining