calreticulin  (Boster Bio)

Journal: Frontiers in Oncology

Article Title: Identification and comprehensive analysis of an immune-related gene prognostic model for indicating tumor immune microenvironment features in soft tissue sarcoma

doi: 10.3389/fonc.2025.1609501

Figure Lengend Snippet: CALR is overexpressed in osteosarcoma and modulates cellular phenotypes. (A) qRT-PCR analysis of CALR mRNA levels in osteosarcoma cell lines (MG-63, 143B, HOS) compared to human normal osteoblasts (hFOB1.19). (B) Western blot showing endogenous CALR protein expression in untreated osteosarcoma cells (Saos-2, MG-63, 143B, U2OS) and CALR-overexpressing 143B cells (OE-CALR). (C) Immunofluorescence staining of CALR (TRITC, red) and nuclei (DAPI, blue) in 143B cells under overexpression (OE-CALR), knockdown (sh-CALR), and negative control (NC) conditions. Fluorescence intensity quantification is shown below. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Cells on coverslips were fixed with 4% PFA, permeabilized with 0.2% Triton X-100, blocked with 3% BSA, and incubated with anti-CALR antibody (#10208-1-AP, 1:200, Proteintech) overnight at 4°C, followed by TRITC-conjugated secondary antibody (1:500) and DAPI (1 μg/mL) at RT.

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Immunofluorescence, Staining, Over Expression, Knockdown, Negative Control, Fluorescence

Journal: International Journal of Oncology

Article Title: Targeting CALR reduces energy metabolism of esophageal cancer cells and inhibits tumor-associated fibroblast infiltration

doi: 10.3892/ijo.2025.5755

Figure Lengend Snippet: CALR is upregulated in ESCC and positively associated with CANX and PDIA3. (A) RNA expression of CALR in ESCC vs. normal specimens in TCGA-ESCA cohort. Correlation between CALR and (B) CANX and (C) PDIA3 at the RNA level in TCGA-ESCA cohort. (D) Immunohistochemical straining of (E) CALR, (F) CANX and (G) PDIA3 expressions in ESCC and normal tissue. Scale bar, 50 µ m. * P<0.05; *** P<0.001 vs. control. CALR, calreticulin; Control, paratumor; ESCC, esophageal squamous cell carcinoma; CANX, calnexin; PDIA3, protein disulfide isomerase A3; ESCA, esophageal carcinoma; TPM, Transcripts Per Million.

Article Snippet: A total of 20 μ g protein/lane was separated by 10% SDS-PAGE, transferred to PVDF membranes and blocked with 5% non-fat dry milk at room temperature for 1 h. The blocked PVDF membranes were incubated overnight at 4°C with primary antibodies against CALR (1:1,000; cat. no. 27298-1-AP; Proteintech Group, Inc.), CANX (1:500; cat. no. BF0515; Affinity Biosciences), PDIA3 (cat. no. 15967-1-AP; Proteintech Group, Inc.), vimentin (cat. no. bs-8533R), N-cadherin (cat. no. bs-1172R), glucose regulatory protein 78 (GRP78) (cat. no. bs-1219R; all BIOSS), α-smooth muscle actin (SMA; cat. no. Bs70000; Biogot Technology Co., Ltd.), fibroblast activation protein (FAP; all 1:1,000; cat. no. bs-5758R; BIOSS), ferroptosis suppressor protein 1 (FSP-1) (1:4,000; cat. no. 20886-1-AP), Platelet-derived growth factor receptors (PDGFR) (cat. no. 13449-1-AP; both Proteintech Group, Inc.), TGF-β (both 1:1,000; cat. no. Ab66043, Abcam).

Techniques: RNA Expression, Immunohistochemical staining, Control

Journal: International Journal of Oncology

Article Title: Targeting CALR reduces energy metabolism of esophageal cancer cells and inhibits tumor-associated fibroblast infiltration

doi: 10.3892/ijo.2025.5755

Figure Lengend Snippet: CALR regulates IP3R calcium ion release channels and ER stress. (A) Protein expression of (B) CALR, (C) CANX and (D) PDIA3 in KYSE150 after treatment with ER stress agonist thapsigargin. (E) Protein expression of (F) CALR, (G) CANX and (H) PDIA3 in KYSE410 cells following treatment with ER stress agonist thapsigargin. (I) In intracellular GRP78 expression after overexpression or knockdown of CALR in KYSE150. (J) Protein expression of GRP78 in KYSE150. (K) In intracellular GRP78 expression after overexpression or knockdown of CALR in KYSE410. (L) Protein expression of GRP78 in KYSE410. (M) Co-IP revealed the interaction of GRP75 and VDAC1 and IP3R1 in KYSE150. (N) Co-IP revealed the interaction of GRP75 and VDAC1 and IP3R1 in KYSE410. †† P<0.01, ††† P<0.001 vs. DMSO, ** P<0.01, *** P<0.001 vs. OE-NC, ### P<0.001 vs. sh-NC. CALR, Calreticulin; IP3R, inositol 1,4,5-Trisphosphate Receptor; ER, endoplasmic reticulum; CANX, calnexin; PDIA3, protein disulfide isomerase A3; ESCC, esophageal squamous cell carcinoma; GRP, glucose regulatory protein; VDAC, voltage-dependent anion channel; IB, immunoblotting; IP, immunoprecipitation; OE, overexpression; NC, negative control; sh, short hairpin.

Article Snippet: A total of 20 μ g protein/lane was separated by 10% SDS-PAGE, transferred to PVDF membranes and blocked with 5% non-fat dry milk at room temperature for 1 h. The blocked PVDF membranes were incubated overnight at 4°C with primary antibodies against CALR (1:1,000; cat. no. 27298-1-AP; Proteintech Group, Inc.), CANX (1:500; cat. no. BF0515; Affinity Biosciences), PDIA3 (cat. no. 15967-1-AP; Proteintech Group, Inc.), vimentin (cat. no. bs-8533R), N-cadherin (cat. no. bs-1172R), glucose regulatory protein 78 (GRP78) (cat. no. bs-1219R; all BIOSS), α-smooth muscle actin (SMA; cat. no. Bs70000; Biogot Technology Co., Ltd.), fibroblast activation protein (FAP; all 1:1,000; cat. no. bs-5758R; BIOSS), ferroptosis suppressor protein 1 (FSP-1) (1:4,000; cat. no. 20886-1-AP), Platelet-derived growth factor receptors (PDGFR) (cat. no. 13449-1-AP; both Proteintech Group, Inc.), TGF-β (both 1:1,000; cat. no. Ab66043, Abcam).

Techniques: Expressing, Over Expression, Knockdown, Co-Immunoprecipitation Assay, Western Blot, Immunoprecipitation, Negative Control

Journal: International Journal of Oncology

Article Title: Targeting CALR reduces energy metabolism of esophageal cancer cells and inhibits tumor-associated fibroblast infiltration

doi: 10.3892/ijo.2025.5755

Figure Lengend Snippet: CALR increases CANX and PDIA3 expression in esophageal squamous cell carcinoma cells. (A) Representative immunohistochemical staining. Relative expression of (B) CALR, (C) CANX and (D) PDIA3 in KYSE150, relative expression of (E) CALR, (F) CANX and (G) PDIA3 in KYSE410 following OE-CALR or sh-CALR transfection. (H) Representative immunofluorescent staining. Scale bar, 100 µ m. Relative expression of (I) CALR, (J) CANX and (K) PDIA3 in KYSE150, relative expression of (L) CALR, (M) CANX and (N) PDIA3 in KYSE410 with OE-CALR or sh-CALR transfection. * P<0.05, ** P<0.01, *** P<0.001 vs. OE-NC, # P<0.05, ## P<0.01 vs. sh-NC. CALR, calreticulin; CANX, calnexin; PDIA3, protein disulfide isomerase A3; OE, overexpression; sh, short hairpin; NC, negative control.

Article Snippet: A total of 20 μ g protein/lane was separated by 10% SDS-PAGE, transferred to PVDF membranes and blocked with 5% non-fat dry milk at room temperature for 1 h. The blocked PVDF membranes were incubated overnight at 4°C with primary antibodies against CALR (1:1,000; cat. no. 27298-1-AP; Proteintech Group, Inc.), CANX (1:500; cat. no. BF0515; Affinity Biosciences), PDIA3 (cat. no. 15967-1-AP; Proteintech Group, Inc.), vimentin (cat. no. bs-8533R), N-cadherin (cat. no. bs-1172R), glucose regulatory protein 78 (GRP78) (cat. no. bs-1219R; all BIOSS), α-smooth muscle actin (SMA; cat. no. Bs70000; Biogot Technology Co., Ltd.), fibroblast activation protein (FAP; all 1:1,000; cat. no. bs-5758R; BIOSS), ferroptosis suppressor protein 1 (FSP-1) (1:4,000; cat. no. 20886-1-AP), Platelet-derived growth factor receptors (PDGFR) (cat. no. 13449-1-AP; both Proteintech Group, Inc.), TGF-β (both 1:1,000; cat. no. Ab66043, Abcam).

Techniques: Expressing, Immunohistochemical staining, Staining, Transfection, Over Expression, Negative Control

Journal: International Journal of Oncology

Article Title: Targeting CALR reduces energy metabolism of esophageal cancer cells and inhibits tumor-associated fibroblast infiltration

doi: 10.3892/ijo.2025.5755

Figure Lengend Snippet: CALR induces epithelial-mesenchymal transition of esophageal squamous cell carcinoma cells. (A) Representative western blots. Relative protein expression of (B) CALR, (C) CANX, (D) PDIA3, (E) vimentin and (F) N-cadherin in KYSE150 with OE-CALR or sh-CALR transfection. Relative protein expression of (G) CALR, (H) CANX, (I) PDIA3, (J) vimentin and (K) N-cadherin in KYSE410 with OE-CALR or sh-CALR transfection. *** P<0.001. CALR, Calreticulin; CANX, calnexin; PDIA3, protein disulfide isomerase A3; OE, overexpression; sh, short hairpin; NC, negative control.

Article Snippet: A total of 20 μ g protein/lane was separated by 10% SDS-PAGE, transferred to PVDF membranes and blocked with 5% non-fat dry milk at room temperature for 1 h. The blocked PVDF membranes were incubated overnight at 4°C with primary antibodies against CALR (1:1,000; cat. no. 27298-1-AP; Proteintech Group, Inc.), CANX (1:500; cat. no. BF0515; Affinity Biosciences), PDIA3 (cat. no. 15967-1-AP; Proteintech Group, Inc.), vimentin (cat. no. bs-8533R), N-cadherin (cat. no. bs-1172R), glucose regulatory protein 78 (GRP78) (cat. no. bs-1219R; all BIOSS), α-smooth muscle actin (SMA; cat. no. Bs70000; Biogot Technology Co., Ltd.), fibroblast activation protein (FAP; all 1:1,000; cat. no. bs-5758R; BIOSS), ferroptosis suppressor protein 1 (FSP-1) (1:4,000; cat. no. 20886-1-AP), Platelet-derived growth factor receptors (PDGFR) (cat. no. 13449-1-AP; both Proteintech Group, Inc.), TGF-β (both 1:1,000; cat. no. Ab66043, Abcam).

Techniques: Western Blot, Expressing, Transfection, Over Expression, Negative Control

Journal: International Journal of Oncology

Article Title: Targeting CALR reduces energy metabolism of esophageal cancer cells and inhibits tumor-associated fibroblast infiltration

doi: 10.3892/ijo.2025.5755

Figure Lengend Snippet: CALR regulates tumor growth and the expression of tumor-associated fibroblast activation marker proteins in mice. (A) Subcutaneous graft. (B) Size of the subcutaneous graft. (C) Staining and (D) quantification of collagen fibers by Masson's staining. (E) Staining and (F) quantification of reticular fibers. (G) Expression of (H) CALR, (I) CANX and (J) PDIA3 detected by immunofluorescent staining. Scale bar, 100 µ m. (K) Expression of (L) CALR, (M) CANX and (N) PDIA3 detected by western blotting. (O) Representative western blots. Expression levels of tumor-associated fibroblast activation marker proteins (P) α-SMA, (Q) FAP, (R) FSP1, (S) PDGFR and (T) TGF-β were detected by western blotting. ** P<0.01, *** P<0.001. CALR, Calreticulin; CANX, calnexin; PDIA3, protein disulfide isomerase A3; SMA, smooth muscle actin; FAP, fibroblast Activation Protein; FSP1, fibroblast specific protein 1; PDGFR, platelet derived growth factor receptors; sh, short hairpin; NC, negative control.

Article Snippet: A total of 20 μ g protein/lane was separated by 10% SDS-PAGE, transferred to PVDF membranes and blocked with 5% non-fat dry milk at room temperature for 1 h. The blocked PVDF membranes were incubated overnight at 4°C with primary antibodies against CALR (1:1,000; cat. no. 27298-1-AP; Proteintech Group, Inc.), CANX (1:500; cat. no. BF0515; Affinity Biosciences), PDIA3 (cat. no. 15967-1-AP; Proteintech Group, Inc.), vimentin (cat. no. bs-8533R), N-cadherin (cat. no. bs-1172R), glucose regulatory protein 78 (GRP78) (cat. no. bs-1219R; all BIOSS), α-smooth muscle actin (SMA; cat. no. Bs70000; Biogot Technology Co., Ltd.), fibroblast activation protein (FAP; all 1:1,000; cat. no. bs-5758R; BIOSS), ferroptosis suppressor protein 1 (FSP-1) (1:4,000; cat. no. 20886-1-AP), Platelet-derived growth factor receptors (PDGFR) (cat. no. 13449-1-AP; both Proteintech Group, Inc.), TGF-β (both 1:1,000; cat. no. Ab66043, Abcam).

Techniques: Expressing, Activation Assay, Marker, Staining, Western Blot, Derivative Assay, Negative Control