Journal: Veterinary Sciences

Article Title: Analysis of Histochemical Characteristics of Submandibular Gland of the Bactrian Camel

doi: 10.3390/vetsci12020108

Figure Lengend Snippet: Immunohistochemical distribution characteristics of EGF and EGFR. ( a ) Immunohistochemical expression of EGF in the submandibular gland of a Bactrian camel; ( b ) immunohistochemical expression of EGFR in the submandibular gland of a Bactrian camel; ( c ) Bactrian camel submandibular gland negative control; ( d ) immunofluorescence expression of EGF in a submandibular gland of the Bactrian camel; ( e ) EGFR immunofluorescence expression in the submandibular gland of a Bactrian camel; ( f ) expression of nuclear immunofluorescence in a submandibular gland of a Bactrian camel; ( g ) EGF and EGFR in the submandibular gland of Bactrian camel immunofluorescence double localization. StD: striated ducts; ID: intercalated ducts (white and black arrow). Scale bars, 20 μm ( a – c ) and 50 μm ( d – g ).

Article Snippet: Rabbit polyclonal antibodies against EGF (Bioss Cat# bs-2008R, RRID:AB_10856305), EGFR (bs-10007R), and an IHC test kit (PV-0023) were purchased from Beijing BIOSS Antibodies, Ltd. (Beijing, China).

Techniques: Immunohistochemical staining, Expressing, Negative Control, Immunofluorescence

Journal: Veterinary Sciences

Article Title: Analysis of Histochemical Characteristics of Submandibular Gland of the Bactrian Camel

doi: 10.3390/vetsci12020108

Figure Lengend Snippet: Distribution of EGF and EGFR in submandibular gland tissues of Bactrian camels detected by immunofluorescence. -: weak positive expression. ++: medium intensity positive expression. +++: strong positive.

Article Snippet: Rabbit polyclonal antibodies against EGF (Bioss Cat# bs-2008R, RRID:AB_10856305), EGFR (bs-10007R), and an IHC test kit (PV-0023) were purchased from Beijing BIOSS Antibodies, Ltd. (Beijing, China).

Techniques: Immunofluorescence, Expressing

Journal: BMC Genomics

Article Title: Identification of candidate gonadal sex differentiation genes in the chicken embryo using RNA-seq

doi: 10.1186/s12864-015-1886-5

Figure Lengend Snippet: RNA-seq expression of novel candidates for in vivo validation

Article Snippet: The FGFR3 antibody was from Bioss (bs-0165R) and used at 1:100.

Techniques: Expressing, In Vivo

Journal: BMC Genomics

Article Title: Identification of candidate gonadal sex differentiation genes in the chicken embryo using RNA-seq

doi: 10.1186/s12864-015-1886-5

Figure Lengend Snippet: Female candidate genes expression in vivo. Whole mount in situ hybridisation for 3 female-biased candidate genes, on E6 UGS from males and females. CAPN5 is more highly expressed in female ( a ) than in males, where only low staining is observed ( g , j ). CAPN5 is also expressed in the adrenal glands in both females and males ( a , g arrows ). In over-stained sections CAPN5 appears to be expressed in the juxta-cortical medulla of the ovary ( d ). GPR56 shows strong expression in female gonads ( b ) and no expression in the males ( h , k ), consistent with RNA-seq. In females it is expressed in the cortex of the ovary, with a lower level in the medulla ( e ). FGFR3 is also higher in females ( c ) than males, which show low expression ( i – gonads delineated by lines). FGFR3 is mostly expressed in the ovarian cortex ( f) , and in the male weakly in a subset of cord cells ( l , arrow heads ). These results are consistent with RNA-seq data. Typically, 3 UGS from each sex were used for each probe, and these images are representative of what is seen. A sense control probe is also tested, and did not show any staining for any of the genes (data not shown)

Article Snippet: The FGFR3 antibody was from Bioss (bs-0165R) and used at 1:100.

Techniques: Expressing, In Vivo, In Situ, Hybridization, Staining, RNA Sequencing Assay

Journal: BMC Genomics

Article Title: Identification of candidate gonadal sex differentiation genes in the chicken embryo using RNA-seq

doi: 10.1186/s12864-015-1886-5

Figure Lengend Snippet: Expression of candidate genes is altered in sex-reversed gonads. Whole mount in situ of E9 UGS from male controls, female controls or fadrozole treated females. The expression of male candidate genes BMPR2 ( a - c ) and ZNF385b ( d - f ) changed dramatically in fadrozole treated embryos ( c , f ), where expression was almost at the same level as untreated males ( a , d ). NZP also had increased expression in treated females ( i ) compared to untreated ( h ), although expression of this gene has generally dropped in control males at this stage ( g ). LAMA1 expression had also dropped in males at E9 ( j ), and only low expression was found in females ( k ). A slight increase in expression was seen in treated females ( l ). CAPN5 was lost from the gonads (dotted lines) but not the adrenal gland of treated females ( o ) compared to control females ( n ) . In the gonads it obtained a similar level of expression to control males ( m ). GPR56, which is absent in males ( p ), was also lost from treated females ( r ) whereas in untreated females it is strongly expressed in the left ovary ( q ). Similarly, fadrozole treatment abolished FGFR3 staining in the gonads ( u ), to a similar level as untreated males ( s ) from normal strong expression in females ( t )

Article Snippet: The FGFR3 antibody was from Bioss (bs-0165R) and used at 1:100.

Techniques: Expressing, In Situ, Staining

Journal: BMC Genomics

Article Title: Identification of candidate gonadal sex differentiation genes in the chicken embryo using RNA-seq

doi: 10.1186/s12864-015-1886-5

Figure Lengend Snippet: Novel candidate female genes are translated in the embryonic gonads. Immunofluorescence staining using antibodies raised in house (CAPN5 and GPR56) or commercial antibodies (FGFR3) in E8 gonads. CAPN5 protein is only weakly expressed in males ( a ), but shows high expression in a subset of cells in the female gonads ( b ). These cells are present in the juxta-cortical medulla, and staining appears cytoplasmic ( c ). GPR56 is expressed in the cortex of the female ( e , f ), where it appears cytoplasmic, although this staining is weak, and GPR56 is weak in male gonads ( d ). FGFR3 is expressed in the male gonads ( g ) and in the female gonads ( h ), where it is strongly expressed in the membrane or cytosol of the germ cells (see insert Gi). Strong staining in the female cortex ( i ) is probably due to germ cell expression, although may also be due to additional cortical cells expressing this receptor

Article Snippet: The FGFR3 antibody was from Bioss (bs-0165R) and used at 1:100.

Techniques: Immunofluorescence, Staining, Expressing

Journal: Frontiers in Genetics

Article Title: Assembly factor for spindle microtubules ( ASPM ) promotes osimertinib resistance in lung cancer by increasing EGFR stability

doi: 10.3389/fgene.2025.1593314

Figure Lengend Snippet: ASPM enhances EGFR resistance in lung cancer cells. (A) Dose‒effect relationship curves of osimertinib for PC-9 cells; (B) Dose‒effect relationship curves of osimertinib for PC-9 OR cells. (C) mRNA expression levels of ASPM and EGFR in PC-9 and PC-9 OR cells. (D) Protein expression of ASPM and EGFR in PC-9 and PC-9 OR cells. The histogram represents the relative gray values of the intracellular EGFR and ASPM proteins, and the data are presented as the means ± standard deviations (means ± SDs, n = 3 fields). (E) Silencing of the ASPM gene and mRNA expression levels of ASPM in PC-9 and PC-9 OR cells. (F) Dose‒effect relationship curves of osimertinib in PC-9 si ASPM cells. (G) Dose‒effect relationship curves of osimertinib in PC-9 OR si ASPM cells. (H) mRNA expression levels of ASPM were detected in PC-9 cells after overexpressing the ASPM gene. (I) Dose‒effect curves of osimertinib after PC-9 overexpression of ASPM . (J) The viability of the cells in each group was assessed via a CCK-8 assay. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: The membrane was blocked with 5% skim milk for 60 min and incubated with primary antibodies, including ASPM (Proteintech, 26223-1-AP) (1:600) and EGFR (Cusabio, CSB-PA005571LA01HU) (1:1000), overnight at 4 °C, followed by 2 h of incubation with HRP-conjugated goat anti-rabbit (Proteintech, SA00001-2) (1:2000) at room temperature.

Techniques: Expressing, Over Expression, CCK-8 Assay

Journal: Frontiers in Genetics

Article Title: Assembly factor for spindle microtubules ( ASPM ) promotes osimertinib resistance in lung cancer by increasing EGFR stability

doi: 10.3389/fgene.2025.1593314

Figure Lengend Snippet: ASPM plays a key role in the stabilization of drug-resistant cell lines. (A) Scatter plot of the correlation between ASPM and EGFR expression; p < 0.05. (B) Silencing of ASPM followed by ASPM and EGFR mRNA expression. (C) Protein expression of EGFR after silencing ASPM. (D) CHX administration at various time points. The right panel represents the relative gray values of the intracellular EGFR proteins, and the data are presented as the means ± standard deviations (means ± SDs, n = 3 fields). (E) IF images of PC-9 and PC-9 OR cells incubated with primary antibodies against EGFR (red) and ASPM (green). Detection was performed using secondary antibodies coupled to Alexa Fluor 488 and Alexa Fluor 555. The cell nuclei were stained with DAPI (blue), Scale = 7.5 μm. (F) mRNA expression levels of ASPM in PC-9 OR cells after silencing of ASPM -2. (G) PC-9 OR cells in the NC group/si ASPM -2 group were stained with propidium iodide (PI), and the DNA content was detected via flow cytometry. The red color represents the theoretical curve that we fit via ModFit software on the basis of the DNA content distribution data of the cells. (H) Statistics of the flow cytometric results in H plots. n = 3, a p value <0.05 was considered statistically significant, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: The membrane was blocked with 5% skim milk for 60 min and incubated with primary antibodies, including ASPM (Proteintech, 26223-1-AP) (1:600) and EGFR (Cusabio, CSB-PA005571LA01HU) (1:1000), overnight at 4 °C, followed by 2 h of incubation with HRP-conjugated goat anti-rabbit (Proteintech, SA00001-2) (1:2000) at room temperature.

Techniques: Expressing, Incubation, Staining, Flow Cytometry, Software

Journal: Frontiers in Genetics

Article Title: Assembly factor for spindle microtubules ( ASPM ) promotes osimertinib resistance in lung cancer by increasing EGFR stability

doi: 10.3389/fgene.2025.1593314

Figure Lengend Snippet: ASPM is significantly upregulated in NSCLC tumor tissues and strongly associated with reduced patient survival. ASPM silencing attenuates PC-9 and PC-9 OR malignant phenotypes, including proliferation and invasion, and sensitizes resistant cells to osimertinib. In addition, inhibiting the expression of ASPM effectively reduces damage to the cell cycle and protein stability of drug-resistant cells, thereby restoring the expression and function of EGFR.

Article Snippet: The membrane was blocked with 5% skim milk for 60 min and incubated with primary antibodies, including ASPM (Proteintech, 26223-1-AP) (1:600) and EGFR (Cusabio, CSB-PA005571LA01HU) (1:1000), overnight at 4 °C, followed by 2 h of incubation with HRP-conjugated goat anti-rabbit (Proteintech, SA00001-2) (1:2000) at room temperature.

Techniques: Expressing

Journal: Journal of Virology

Article Title: Zika Virus Hijacks Stress Granule Proteins and Modulates the Host Stress Response

doi: 10.1128/JVI.00474-17

Figure Lengend Snippet: ZIKV infection does not induce robust SG formation in A549 cells or primary human fetal astrocytes (HFAs). A549 cells, primary HFAs, and Huh-7 cells were infected with ZIKV strain PLCal or MR766 (MOI of 3) for the indicated time periods and then processed for confocal microscopy analyses. Infected cells were detected using mouse anti-flavivirus E protein antibody; SGs were identified by staining for endogenous G3BP1 and TIAR using rabbit and goat polyclonal antibodies, respectively. (A) Representative images of mock- and ZIKV-infected cells at 48 h.p.i. are shown. (B) Quantification of SGs was performed using the software Volocity, and the average numbers of SGs per cell are shown in the table. A minimum of 30 cells was used for each sample. n.d., not detected. (C) A549 cells were infected with ZIKV (PLCal strain) for 48 h and then processed for confocal microscopy analyses. P-bodies were identified by staining Dcp1a with a rabbit polyclonal antibody. Primary antibodies were detected with donkey anti-mouse IgG conjugated to Alexa Fluor 546, donkey anti-rabbit IgG conjugated to Alexa Fluor 488, and chicken anti-goat IgG conjugated to Alex Fluor 647. Nuclei were stained with DAPI. Size bars are 17 μm (A) and 100 μm (C). Images were acquired on a spinning-disk confocal microscope. (D) Quantification of P-bodies was performed using Volocity software. A minimum of 30 cells were used for each sample. Values are expressed as the average number of P-bodies per cell ± standard errors from three independent experiments.

Article Snippet: Purified ZIKV NS5 was used to generate goat polyclonal antibody (ProSci Incorporated).

Techniques: Infection, Confocal Microscopy, Staining, Software, Microscopy