pax2  (Addgene inc)

Journal: Experimental & Molecular Medicine

Article Title: Deficiency of exocyst complex component Exoc5 exacerbates the progression of kidney fibrosis

doi: 10.1038/s12276-026-01649-8

Figure Lengend Snippet: a Western blot of Pax2 and PCNA expression. b , c , Graphs of Pax2 ( b ) and PCNA ( c ) expression in the kidney tissue; the densities of bands were quantified using ImageJ software ( n = 6). GAPDH was used as the loading control. d , f , h The kidney sections were subjected to immunofluorescence staining using antibodies against Pax2 (green) and AQP1 (red) ( d ), PCNA (green) and AQP1 (red) ( f ), BrdU (green) and AQP1 (red) ( h ). DAPI staining (blue) was performed to visualize cell nuclei. Arrowheads indicate Pax2-, PCNA- and BrdU-positive nuclei in proximal tubule cells, respectively. e , g , i The quantification was performed by counting Pax2- ( e ), PCNA- ( g ) and BrdU-positive cells ( i ) in proximal tubules in a 40× micrograph with a field area of 0.1 mm 2 . j , l The kidney sections were subjected to immunofluorescence staining using antibodies against Pax2 (green) and PCNA (red) ( j ) and Pax2 (red) and BrdU (green) ( l ). DAPI staining (blue) was used to visualize cell nuclei. k , m The quantification was performed by counting Pax2- and PCNA-positive cells ( k ) and Pax2- and BrdU-positive cells ( m ) in proximal tubules in a 40× micrograph with a field area of 0.1 mm 2 . Pax2-, PCNA- and BrdU-positive cells in proximal tubules are marked by arrowheads, as indicated by the colors in the figures. PT–Exoc5 WT and PT–Exoc5 KO mice were subjected to either UUO or sham surgery, and kidneys were collected 7 days following the surgery. BrdU was administered every other day from the day before UUO until death. Results are expressed as the mean ± s.e.m. ( n = 4–6). * P < 0.05. NS, not significant; ND, not detected.

Article Snippet: Antibodies against the following proteins were used: Exoc5 (cat. no. sc-514802), Exoc4 (cat. no. VAM-SV016, StressMarq Biosciences), Exoc6 (cat. no. 12723-1-AP, Proteintech), Exoc7 (cat. no. 28666, Cell Signaling Technology), p-Smad3 (cat. no. ab52903, Abcam), Smad3 (cat. no. 9523, Cell Signaling Technology), β-catenin (cat. no. 8480, Cell Signaling Technology), alpha-smooth muscle actin (α-SMA; cat. no. A2547, Sigma-Aldrich), vimentin (cat. no. 5741, Cell Signaling Technology), Pax2 (cat. no. PRB-276P, Covance), proliferating cell nuclear antigen expression (PCNA; cat. no. m879, DAKO), YAP/TAZ (cat. no. 8418, Cell Signaling Technology), YAP (cat. no. 14074, Cell Signaling Technology), phosphorylated YAP (p-YAP; cat. no. 13008, Cell Signaling Technology), CTGF (cat. no. sc-365970), CYR61 (cat. no. 39382, Cell Signaling Technology), N-cadherin (cat. no. 13116, Cell Signaling Technology) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH; cat. no. NBP600-502, NOVUS).

Techniques: Western Blot, Expressing, Software, Control, Immunofluorescence, Staining

Journal: Experimental & Molecular Medicine

Article Title: Deficiency of exocyst complex component Exoc5 exacerbates the progression of kidney fibrosis

doi: 10.1038/s12276-026-01649-8

Figure Lengend Snippet: a – k Western blot ( a , d and i ) of EXOC5 ( b and e ), PAX2 ( c ), EXOC4 ( f ), EXOC6 ( g ), EXOC7 ( h ), YAP ( j ) expression in the cell lysate; band densities were quantified using ImageJ software ( n = 3–4); The p-YAP/YAP ratio was calculated on the basis of band intensity ( k ). l The fixed cells were subjected to immunofluorescence staining using anti-YAP antibody (green). m – o Western blot ( m ) of the vimentin ( n ) and N-cadherin ( o ) expression in the cell lysate, analyzed 48 h after TGF-β treatment, and band densities were quantified using ImageJ software ( n = 4). GAPDH was used as a loading control. p , q The fixed cells were subjected to immunofluorescence staining using anti-α-SMA antibody (red) ( p ), and α-SMA-positive cells were counted ( q ) in a 20× micrograph. DAPI staining (blue) was used to visualize cell nuclei. HK-2 cells were transfected with either Exoc5-siRNA or scrambled siRNA and the cells were treated with or without 5 ng/ml TGF-β for 1 or 48 h. Results are expressed as the mean ± s.e.m. ( n = 3). * P < 0.05. NS, not significant.

Article Snippet: Antibodies against the following proteins were used: Exoc5 (cat. no. sc-514802), Exoc4 (cat. no. VAM-SV016, StressMarq Biosciences), Exoc6 (cat. no. 12723-1-AP, Proteintech), Exoc7 (cat. no. 28666, Cell Signaling Technology), p-Smad3 (cat. no. ab52903, Abcam), Smad3 (cat. no. 9523, Cell Signaling Technology), β-catenin (cat. no. 8480, Cell Signaling Technology), alpha-smooth muscle actin (α-SMA; cat. no. A2547, Sigma-Aldrich), vimentin (cat. no. 5741, Cell Signaling Technology), Pax2 (cat. no. PRB-276P, Covance), proliferating cell nuclear antigen expression (PCNA; cat. no. m879, DAKO), YAP/TAZ (cat. no. 8418, Cell Signaling Technology), YAP (cat. no. 14074, Cell Signaling Technology), phosphorylated YAP (p-YAP; cat. no. 13008, Cell Signaling Technology), CTGF (cat. no. sc-365970), CYR61 (cat. no. 39382, Cell Signaling Technology), N-cadherin (cat. no. 13116, Cell Signaling Technology) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH; cat. no. NBP600-502, NOVUS).

Techniques: Western Blot, Expressing, Software, Immunofluorescence, Staining, Control, Transfection